CN105688029A - A pharmaceutical composition containing a composite extract product of saururus chinensis, turmeric and milkwort root and used for preventing or treating obesity - Google Patents

A pharmaceutical composition containing a composite extract product of saururus chinensis, turmeric and milkwort root and used for preventing or treating obesity Download PDF

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Publication number
CN105688029A
CN105688029A CN201410710686.3A CN201410710686A CN105688029A CN 105688029 A CN105688029 A CN 105688029A CN 201410710686 A CN201410710686 A CN 201410710686A CN 105688029 A CN105688029 A CN 105688029A
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China
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rhizoma
extract
saururi
radix polygalae
experimental group
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马珍烈
郭东勳
李智慧
金童乾
金兑洙
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KOREA INST OF ORIENTAL MEDICIN
Korea Institute of Oriental Medicine KIOM
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KOREA INST OF ORIENTAL MEDICIN
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Abstract

The invention relates to a pharmaceutical composition and a food composition which are used for preventing or treating obesity. The compositions contain a composite extract product of saururus chinensis, turmeric and milkwort root or a composite extract product of saururus chinensis, turmeric, milkwort root and grassleaf sweetflag rhizome. The invention also relates to a preparing method of the composite extract products. The compositions containing the composite extract product can hinder lipogenesis under the conditions of no cytotoxicity, and therefore the compositions can be widely used for preventing, improving or treating the obesity or obesity related diseases.

Description

The pharmaceutical compositions for preventing or treat obesity of the composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae
Technical field
The present invention relates to the compositions for preventing or treat obesity of a kind of composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, more particularly it relates to an the preparation method for preventing or treat the pharmaceutical compositions of obesity, food compositions and described composite extract of the composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae or the composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
Background technology
Generally, the fat state representing that body fat tissue is too much, is phenomenon that is unbalanced with the energy consumed by body movement etc. by the energy of food intake and that make remaining energy stacking be body fat。If unbalanced and cause body fat abnormal increase through long energy, could various metabolic diseases and the adult diseases such as induced Diabetic, hyperlipidemia, heart disease, apoplexy, arteriosclerosis, fatty liver。This phenomenon not only exists in west, also becomes serious social problem in Korea S。
The a variety of causes such as psychological factor that environmental factors that hereditary factors, westernization dietetic life bring, pressure bring, abnormal energy metabolism can cause obesity。It addition, its kind is divided into due to gluttony according to reason and lacks kinetic Obesity and the disease caused due to incretion, hypothalamic, heritability, metabolic etc. is fat。
Evaluate the method that the method for bluntness has the method measuring body weight, measures wrinkle of skin thickness, be generally more than 25 be defined as obesity by bluntness index。Body obesity index (constitutional index, bodymassindex, BMI) is the body weight () square value drawn divided by height (m)。For westerner, bluntness be more than 30 be evaluated as obesity, however, it is contemplated that the difference between race, in Korea S, by bluntness be more than 25 be evaluated as obesity。
In order to develop Bariatric agent, the whole world is just carrying out many-sided research。Bariatric medicine substantially can be divided into suppression fat absorption, reduces fat and promote heat generation, modulation of appetite and satiety, obstruction protein metabolism and the mood regulation mechanism relevant to food intake。Representational Bariatric agent has using orlistat (oristat) as raw material to suppress the orlistat of fat absorptionTM(XenicalTM) and stimulate sympathetic nervous system using sibutramine (sibutramine) as main material, thus the Reductil of appetite-suppressingTM(ReductilTM)。But, for orlistatTM, report that it has the side effect such as steatorrhea, abdominal pain, vomiting, pruritus, hepatic injury, for ReductilTM, not only there is due to it side effect such as headache, inappetence, insomnia, constipation, but also serious cardiovascular side effects can be caused, therefore, cause the arguements such as strengthening use standard recently。
Except having the pharmacotherapy by this Bariatric agent, as the exercise therapy of prevention and the dietetic therapy of the method also restricted food intake for the treatment of obesity, increase energy expenditure, and also implement psychotherapy, action therapy, surgical treatment etc.。
As preferred Bariatric method, it has been disclosed that the most safely and effectively method be promote energy expenditure by moving while, the method carrying out being used in combination with the Bariatric medicine of few side effects。But, orlistatTMAnd ReductilTMIt is in the news serious side effect Deng Bariatric medicine, in safety, can not get clear and definite trust。Accordingly, it would be desirable to while developing a kind of anti-obesity effect that human body performance is excellent, security performance accesses the material of guarantee。
In this case, the present inventor strives to find and human body does not cause side effect, and the result of the material of Bariatric excellent effect, obesity is shown prevention and the effect for the treatment of by the composite extract confirming Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae in the scope that cell does not produce toxicity, thus completing the present invention。
Summary of the invention
Solve the technical problem that
It is an object of the invention to, it is provided that the composite extract of a kind of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae or its fraction are used for the application treating in the medicine of obesity in preparation。
Described composite extract is that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are with 1:(0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w)。Described composite extract can be the composite extract by utilizing the ethanol (EtOH) of 25% to obtain。
Another object of the present invention is to, it is provided that the composite extract of a kind of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei or its fraction are used for the application treating in the medicine of obesity in preparation。
Described composite extract can be that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are with 1:(0.2～1): (0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w/w)。Described composite extract can be the composite extract by utilizing the ethanol (EtOH) of 25% to obtain。
Another object of the present invention is to, it is provided that the preparation method of the composite extract of a kind of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。
Described test portion can with 1:(0.2～1): the ratio of (0.2～1) (w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
Another object of the present invention is to, it is provided that the preparation method of the composite extract of a kind of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei, described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。
Described test portion can with 1:(0.2～1): (0.2～1): the ratio of (0.2～1) (w/w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
Another object of the present invention is to, it is provided that a kind of compositions, described compositions contains composite extract or its fraction of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
Described composite extract can be that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are with 1:(0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w)。Described compositions can be the compositions with Bariatric purposes。Described composite extract can be the composite extract by utilizing the ethanol (EtOH) of 25% to obtain。
Another object of the present invention is to, it is provided that a kind of compositions, described compositions contains composite extract or its fraction of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
Described composite extract can be that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are with 1:(0.2～1): (0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w/w)。Described compositions can be the compositions with Bariatric purposes。Described composite extract can be the composite extract by utilizing the ethanol (EtOH) of 25% to obtain。
Technical scheme
Make an embodiment for achieving the above object, the present invention provides a kind of pharmaceutical compositions for preventing or treat obesity, described pharmaceutical compositions contains the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei or their fraction。
The term " Rhizoma Saururi (Herba Saururi) (SaururichinensisBaill.) " of the present invention represents the perennial herb of the Piperales Saururaceae used in the traditional Chinese medical science as Chinese medicine。Known described Rhizoma Saururi (Herba Saururi) bitter in the mouth and pungent；Flower and root are white；When blooming, 2～3 in upper leaves become white；Alabastrum is respectively born the fruit of a circle when 9～October。
According to the present invention, described Rhizoma Saururi (Herba Saururi) can use as the effective ingredient being used for preventing or treating the pharmaceutical compositions of obesity, and the position at this moment used can be the whole position of leaf, stem, root or plant。
The term " Rhizoma Curcumae Longae (CurcumaeLongaeRhizoma) " of the present invention represents the perennial herb of the Rhizoma Zingiberis Recens order Zingiberaceae used in the traditional Chinese medical science as Chinese medicine。Known described Rhizoma Curcumae Longae bitter in the mouth and peppery；The outer surface of root and stem is faint yellow, internal in salmon pink；Distribute Camphora fragrance；Spica is Zao more raw than leaf；Axil in 4～June place grows yellow flowers。
According to the present invention, described Rhizoma Curcumae Longae can use as the effective ingredient being used for preventing or treating the pharmaceutical compositions of obesity, and the position at this moment used can be stem or root position。
The term " Radix Polygalae (PolygalaeRadix) " of the present invention represents the perennial herb of the cattle Seedling order Polygalaceae used in the traditional Chinese medical science as Chinese medicine。Known described Radix Polygalae taste is peppery and bitter；Root is thick and long in carinate；Almost without hair except top has the hair being slightly curved；Purple flower is opened during 7～August；Fruit is divided into multiple grid and is referred to as the capsule containing multiple seeds。
According to the present invention, described Radix Polygalae can use as the effective ingredient being used for preventing or treating the pharmaceutical compositions of obesity, and the position at this moment used can be root position。
The term " Rhizoma Acori Graminei (AcoriGramineriRhizoma) " of the present invention represents the perennial herb of the Arales Araeceae used in the traditional Chinese medical science as Chinese medicine。The root of known described Rhizoma Acori Graminei and stem outwards stretch；Tuberosity place grows fibrous root；Distance between each tuberosity in ground, and it is short to expose the spacing between the tuberosity on earth's surface；Overall in green；Flower is the hermaphrodite flower with gynoecium and stamen；Yellow flowers is opened during 6～July；Fruit is divided into multiple grid and has a lot of seed。
According to the present invention, described Rhizoma Acori Graminei can use as the effective ingredient being used for preventing or treating the pharmaceutical compositions of obesity, and the position at this moment used can be stem or root position。
The composite extract that test portion containing described Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei maybe is carried out extracting and obtains by the composite extract that the test portion containing described Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae is carried out extracting and obtains by term " composite extract " expression of the present invention。Difference according to extract part, each extract of plant or the activity of its fraction whether, level of activity and contained therein concrete become branch different。The present inventor finds out that Rhizoma Saururi (Herba Saururi) plant is overall at first, the composite extract of the root position of the stem of Rhizoma Curcumae Longae and root and Radix Polygalae, or the effect that Rhizoma Saururi (Herba Saururi) plant is overall, the stem of Rhizoma Curcumae Longae and the composite extract of root position, the root position of Radix Polygalae and the stem of Rhizoma Acori Graminei and root position demonstrate prevention or treatment is fat。
Further, described composite extract is not only proved to as obesity has prevention or therapeutic effect, and, by the experimental result to mice, its stability have also been obtained proof (Fig. 7 and Fig. 8)。On the other hand, the composition of composite extract is constituted, for instance, the individuality of Radix Polygalae extract shows great toxicity (embodiment 3-2, Fig. 4), i.e. in hepatic tissue and renal tissue, show great chronic inflammatory disease (Fig. 7 and Fig. 8)。This result implys that the individual components of the composite extracts such as Radix Polygalae extract is difficult to individually be administered to individuality。
According to the present invention, described composite extract can use as the effective ingredient being used for preventing or treating the pharmaceutical compositions of obesity, as long as the content for obtaining Rhizoma Saururi (Herba Saururi) contained in the test portion of described composite extract, Rhizoma Curcumae Longae and Radix Polygalae can when using in demonstrating the preparation of compositions of prevention or the fat effect for the treatment of, it does not have limits especially。But the mixing ratio of the Rhizoma Saururi (Herba Saururi) contained in described test portion, Rhizoma Curcumae Longae and Radix Polygalae can be preferably 1:(0.2～1): (0.2～1) (w/w/w), it is more preferably 1:(0.4～1): (0.4～1) (w/w/w), it is most preferred that for 1:0.6:1 (w/w/w)。
In addition, when obtaining composite extract from the above-mentioned test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei, as long as the content of Rhizoma Saururi (Herba Saururi) contained in described test portion, Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei can use in the preparation demonstrating the compositions of effect of prevention or treatment obesity, it does not have limit especially。But the mixing ratio of the Rhizoma Saururi (Herba Saururi) contained in described test portion, Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei is preferably 1:(0.2～1): (0.2～1): (0.2～1) (w/w/w/w), it is more preferably 1:(0.4～1): (0.4～1): (0.4～1) (w/w/w/w), it is most preferred that for 1:0.6:1:0.4 (w/w/w/w)。And it is possible to use commercial sale or from naturally gathering or the described Rhizoma Saururi (Herba Saururi) of cultivation, Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
For described extracting method, it is preferred to use boiled water extraction (), hot water extraction, Soakage extraction, reflux cooling extract or ultrasonic extraction, but be not limited to this。
Described extract can use Extraction solvent to extract or add fractional distillation solvent in the extract prepared by using Extraction solvent to extract to carry out fractional distillation and prepare。For described Extraction solvent, it is possible to use water, organic solvent or their mixed solvent etc., but it is not limited to this。For described organic solvent, it is possible to use carbon number is alcohol or the non-polar solven such as ethyl acetate or acetone polar solvent, hexane or dichloromethane of 1 to 4, or can use their mixed solvent。And, it is preferable that the alcohol that water, carbon number can be used to be 1 to 4 or their mixed solvent, more preferably can use ethanol。By utilizing the ethanol (EtOH) of 25% to be prepared for extract as described solvent in one embodiment of the invention。
Gross weight gauge with pharmaceutical compositions, it is possible to contain the described extract of 0.001 to 100 weight % respectively, it is preferable that the described extract of 0.1 to 80 weight % can be contained respectively。
The term " fraction of composite extract " of the present invention represents by separating the result thing that the fractional method of special component or special groups obtains from described each composite extract。
In the present invention, described fraction can obtain by composite extract is suitable for multiple fractional method, solvent fractionation that described fractional method can be implemented for multi-solvents is processed, by having ultrafiltration fractionating process that the ultrafilter membrane of the molecular weight cutoff specified implements, implement the chromatography fractionating process etc. of plurality of color Zymography (in order to prepare) according to size, electric charge, hydrophobicity, the separation of affinity, but be not limited to this。For the solvent used in described solvent fractionation, it is possible to use polar solvent or non-polar solven, it is preferred to use non-polar solven, but it is not limited to this。Described solvent fractionation can from nonpolar high to Low solvent, the method for fractional distillation successively be implemented by using, for instance, it is possible to use by using hexane or ethyl acetate and the method for composite extract described in fractional distillation successively。
Gross weight gauge with pharmaceutical compositions, it is possible to contain the described fraction of 0.001 to 100 weight % respectively, it is preferable that the described fraction of 0.1 to 80 weight % can be contained respectively。
The term " pharmaceutical compositions " of the present invention represents the compositions of preparation for the purpose of prevention or treatment disease, various form can be turned to according to conventional method dosage form and use respectively, such as, dosage form can turn to the peroral dosage forms such as powder, granule, tablet, capsule, suspension, emulsion, syrup, it is also possible to dosage form turns to the form of external preparation, suppository and sterilizing injecting solution and uses。
Health is caused unsuitable state owing to body weight increases by term " obesity " expression of the present invention, it is also represented by owing to caloric intake specific energy consumption is many, so there is no be consumed and remaining energy piles up, with the form of multiple lipid, the state raised for lipid content in body fat or blood。If it occur that above-mentioned obesity, various metabolic diseases and the adult diseases such as diabetes, hyperlipidemia, heart disease, apoplexy, arteriosclerosis, fatty liver can be caused。
Compositions provided by the invention, it is possible not only to suppress lipogenesis to suppress fat, and can improve, prevent or treat from the derivative obesity-related disease of obesity, for instance, the various metabolic diseases such as diabetes, hyperlipidemia, heart disease, apoplexy, arteriosclerosis, fatty liver。
Term " prevention " in the present invention represents by being administered all behaviors that the compositions of the present invention suppresses or postpone the morbidity of described obesity or obesity-related disease。
Term " treatment " in the present invention represents by being administered all behaviors that the compositions of the present invention makes described obesity or obesity-related disease take a turn for the better or change to useful direction。
According to one embodiment of the invention, obtain respectively containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, the test portion of Radix Polygalae and Rhizoma Acori Graminei or hot water extract's (composite extract or indivedual extract) of indivedual test portion containing described each composition, warm water extraction thing (composite extract or indivedual extract) or ethanol extraction (composite extract or indivedual extract) (referring to table 1), and carry out processing (referring to table 2) and identifying the result of In Vitro Anti obesity effect in 3T3-L1 by multiple concentration by described each extract, confirm as hot water extract (Fig. 1 a to Fig. 1 c), in warm water extraction thing (Fig. 2 a to Fig. 2 c) or ethanol extraction (Fig. 3 a to Fig. 3 c), composite extract and Radix Polygalae extract show fat content and reduce effect and cell quantity minimizing effect, Rhizoma Saururi extract, Rhizoma Curcumae Longae extract and Rhizoma Acori Graminei extract reduce effect and cell quantity minimizing effect entirely without showing fat content。Further, in described hot water extract, warm water extraction thing and ethanol extraction, the fat content minimizing effect & Safety confirming as ethanol extraction is the most excellent。And, described each extract is identified the result of the In Vitro Anti obesity effect of described each extract to mice administration (referring to table 3) with multiple content nursing food rich in fat, and the mice of administration Radix Polygalae extract and composite extract (ethanol extraction) shows the most excellent effect of losing weight。Wherein, the toxicity that Radix Polygalae extract performance is serious, in contrast, the toxicity confirming as composite extract (ethanol extraction) weakens (referring to Fig. 4)；Carrying out fat weight measurement result, analyze as being Radix Polygalae to the composition suppressing fat to increase influential effect maximum, be secondly Dioscorea zingiberensis, the inhibition that fat is increased by Rhizoma Acori Graminei contributes minimum (referring to Fig. 6)；Confirm as composite extract (ethanol extraction) in hepatic tissue (referring to Fig. 7) and renal tissue (referring to Fig. 8) and show higher safety than Radix Polygalae extract。
According to another embodiment of the present invention, in order to confirm other material except analyzing, in the composition contained by described composite extract, fat being increased the Rhizoma Acori Graminei that inhibition is minimum, namely the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae is to fat inhibition, Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are contained as essential component, and optionally contains Rhizoma Acori Graminei and trefoil test portion and obtain ethanol extraction (referring to table 4) as object respectively。Each ethanol extraction of described acquisition is obtained laboratory animal (referring to table 5) with multiple consumption respectively to mice administration, using the laboratory animal of described acquisition as object, body weight (referring to Fig. 9), food intake (referring to Figure 10), fat weight (Figure 11 a to Figure 11 d), hungry level (referring to Figure 12 a to 12c), leptin level (referring to Figure 13) and blood constituent rank (referring to Figure 14 a to 14k) are evaluated。Itself it was found that not only the composite extract of Rhizoma Saururi (Herba Saururi) Rhizoma Curcumae Longae and Radix Polygalae show the obesity effect of raising compared with matched group, and the composite extract containing Rhizoma Acori Graminei further also shows the obesity effect of raising compared with matched group。
As a result it will be appreciated that composite extract contained in the obesity compositions of the present invention can available from the test portion that must contain Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, or can available from the test portion containing Rhizoma Acori Graminei further。
The pharmaceutical compositions of the described present invention can also contain pharmaceutically acceptable diluent, excipient or carrier further。Described compositions containing pharmaceutically acceptable carrier can be the multiple dosage forms such as oral or non-per os。Time formulation, use normally used filler, diluent (), prepared by binding agent, wetting agent, disintegrating agent, the diluent such as surfactant or excipient。Solid preparation as oral administration includes tablet pill, powder, granule, capsule etc.。These solid preparations by mixing more than one excipient in more than one compound, for instance, starch, calcium carbonate, sucrose, lactose, gelatin etc. and prepare。Further, except merely using excipient, the lubricant such as magnesium stearate, Talcum is also used。Suspending agent, mixture for internal use (), Emulsion, syrup etc. belong to oral liquid preparation。Oral liquid preparation can include commonly using as beyond the water of single diluent, liquid Paraffin, it is also possible to include multiple excipient, for instance, wetting agent, sweeting agent, aromatic, preservative etc.。Non-oral administration preparation includes sterile water solution, non-aqueous solvent, suspending agent, Emulsion, freeze-dried preparation, suppository。The ester etc. that propylene glycol, Polyethylene Glycol, Fructus Canarii albi wet goods plant oil, ethyl oleate etc. can be injected can be used to be used as non-aqueous solvent or suspended solvents uses。Witepsol, Polyethylene Glycol (Macrogol), tween (tween) 61, cocoa butter, bright mesophyll reality, glycerin jelly etc. can serve as the base material of suppository。
Described pharmaceutical compositions can be the arbitrary dosage form in tablet, pill, powder, granule, capsule, suspending agent, mixture for internal use, Emulsion, syrup, sterile water solution, non-aqueous solvent, suspending agent, Emulsion, freeze-dried preparation and suppository。
Another embodiment as the purpose in order to reach the invention described above, the present invention provides a kind of prevention or the fat method for the treatment of, and described method includes to be used for, as effective ingredient, the step that the pharmaceutical compositions preventing or treat obesity is administered to individuality that is that have fat probability or that have occurred and that obesity containing described composite extract or its fraction。At this moment, each effective ingredient being included in described composite extract is described above。
Term " individuality " in the present invention represents and includes having occurred and that obesity or it may happen that all animals of people of obesity。
Term " administration " in the present invention represents the behavior of the pharmaceutical compositions being imported the present invention by suitable method to individuality。
In the present invention, the administration of described pharmaceutical compositions, as long as tissue can be achieved the goal, it is possible to implemented by general any approach。The pharmaceutical compositions of the present invention can pass through the method administrations such as oral administration, oral administration, rectally, topical, intraperitoneal administration, intravenous administration, intraarterial delivery, intramuscular adminstration, intranasal administration, subcutaneous administration, intradermal administration, percutaneous dosing, intranasal administration, feeding drug into pulmones, drop rectum with drug, eyeball administration according to purpose, but is not limited to this。Further, described pharmaceutical compositions can use active substance can be administered to any device of purpose signaling。
Pharmaceutical compositions according to one embodiment of the invention contains described composite extract。With pharmaceutical compositions gross weight gauge, containing the described composite extract of 0.1 to 50 weight % content, but it is not limited to this。
The described pharmaceutical compositions of the present invention can pharmaceutically effectively to measure administration。Term in the present invention " pharmaceutically effectively amount " represents that the ratio can be applicable to reasonably being benefited of therapeutic treatment/dangerous is for treating the sufficient amount of disease, the degree of effective dose according to include individual kind and severity, the age, sex, pharmaceutically active, to the sensitivity of medicine, administration time, route of administration and eliminating ratio, treatment time, the factor of medicine used and other factor decision known in medical domain simultaneously。The pharmaceutical compositions of the present invention can as independent Therapeutic Administration or with other therapeutic agent coupling be administered, it is possible to be administered successively with known treatment agent or be administered simultaneously。Furthermore it is possible to single administration or multiple dosing。Considering described factor, it is most important for obtaining maximum effect with minimum amount administration under the degree not having side effects。
The dosage of pharmaceutical compositions for preventing or treat obesity of the present invention, those skilled in the art can according to application target, the severity of disease, the age of patient, body weight, sex, medical history or determine as the kind etc. of material of effective ingredient use。As an embodiment, it is possible to the pharmaceutical compositions of the present invention is administered with each adult about 1 μ dose of g/kg/ days to 100mg/kg/ days, it is preferred to 20 to 30mg/kg/ sky。The administration frequency of described pharmaceutical compositions is not made particular determination, it is possible to be divided into multiple dosing once a day or by consumption。
As realizing another embodiment of described purpose, the present invention provides a kind of food compositions for preventing or improve obesity, and described compositions contains described composite extract or its fraction。
Owing to the Rhizoma Saururi (Herba Saururi) contained in described composite extract, Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are used as Chinese crude drug for a long time, therefore, it is possible to prove its safety, such that it is able to often eat, it is possible to so that the form of the food of prevention or improvement obesity can be helped to prepare and absorb。At this moment, the content of the described extract contained in described food or its fraction being not particularly limited, with the gross weight gauge of food compositions, the content of described complex composition can be 0.01 to 100 weight %, it is preferred to 1 to 80 weight %。When food is beverage, in 10ml, it is possible to the described composite extract of the ratio containing 1 to 30g, it is preferable that the described composite extract of the ratio of 3 to 20g can be contained。Further, described compositions can contain further be typically in food compositions use the supplementary element that can improve abnormal smells from the patient, taste, vision etc.。For example, it is possible to containing vitamin A, C, D, E, B1, B2, B6, B12, nicotinic acid, biotin, folic acid, pantothenic acid etc.。And, it is also possible to containing mineral such as zinc, ferrum, calcium, chromium, magnesium, manganese, copper。And, it is also possible to containing aminoacid such as lysine, tryptophan, cysteine, valines。And also preservative (calcium sorbate can be added, sodium benzoate, salicylic acid, dihydrokainic acid sodium etc.), antibacterial (bleaching powder and dense bleaching powder, sodium hypochlorite etc.), antioxidant (butylhydroxy anisole, butylated hydroxytoluene etc.), coloring agent (tar color etc.), developer (sodium nitrite, sub-sodium acetate etc.), bleach (sodium sulfite), flavouring agent (monosodium glutamate (MSG), sodium glutamate etc.), sweetening material (dulcin, sucdrol, saccharin, sodium etc.), spice (vanillin, lactone etc.), extender (Alumen, D-potassium hydrogen tartrate etc.), hardening agent, emulsifying agent, thickening agent (thickener), liniment, natural gum bodying agent (), foam in hibitors, solvent, the food additives (foodadditives) such as modifying agent。Described additive is chosen according to the kind of food, and uses with suitable amount。
It addition, utilize the food compositions containing described composite extract or its fraction, it is possible to preparation is for preventing or improve the functional food of obesity。
Specifically such as, described food compositions is used can to prepare the processed food for preventing or improve obesity。This processed food includes cookies, beverage, drinks, fermented food, canned food, milk processed food, processed meat food, noodles etc.。Cookies includes plasticity cookies, sunken cake, cake, bread, sugar, fruit jelly, chewing gum, cereal (include grain corn sheet etc. and replace the kind of staple foods) etc.。Beverage includes drinking water, soda pop, functional ion beverage, fruit juice (such as, Fructus Mali pumilae, Fructus Vitis viniferae, Aloe, mandarin orange, Fructus Persicae, Radix Dauci Sativae, tomato fruit juice etc.), happy dew etc.。Drinks includes rice wine, whiskey, liquor, medicated beer, foreign wine, fruit wine etc.。Fermented food includes soy sauce, salty sauce, Fructus Capsici sauce etc.。Canned food includes Aquatic product thing canned food (such as, tuna, blue and white fish, saury, Carnis Rapanae thomasianae canned food etc.), livestock products canned food (beef, Carnis Sus domestica, Carnis Gallus domesticus, turkey canned food etc.), primary product canned food (Semen Maydis, Fructus Persicae, pineapple tin etc.)。Milk processed food include cheese, butter, Yoghourt etc., processed meat food includes fried pork chop, fried beefsteak, Deep-fried chicken joints, sausage, pot bag meat, cube meat class (), dried beef slices ( ) etc.。Also include the noodles such as seal-packed dough。In addition, described compositions may be used for retort pouch food, soup class etc.。
Term " functional food (functionalfood) " in the present invention is and specific food (foodforspecialhealthuse for health care, FoSHU) identical term, represent except supply nutrition, it is also represented by being processed as the food that the medical science that makes organism regulatory function effectively show, medical effect are excellent, and in order to obtain the beneficial effect for preventing or improve osteopathia, described food can be prepared as the variforms such as tablet, capsule, powder, granule, liquid, pill。
As realizing another embodiment of described purpose, the preparation method that the present invention provides a kind of described composite extract, described composite extract is the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。
In the preparation method of above-mentioned composite extract, for described extracting method, the extracting method commonly used in the art such as hot water extraction method, warm water extraction method, ethanol extraction method, ultrasonic extraction, Filtration and reflux extraction can be used, after obtaining extract, it is also possible to farther include described extract to be concentrated or cryodesiccated step。
As another embodiment realizing described purpose, the preparation method that the present invention provides a kind of composite extract, described composite extract is the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei, described preparation method include by containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae test portion utilize water, C1～C4Alcohol or its mixed solvent carry out extracting and obtain the step of extract。At this moment, each effective ingredient being included in described composite extract is described above。
Described test portion can with 1:(0.2～1): the ratio of (0.2～1) (w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
As another embodiment realizing described purpose, the preparation method that the present invention provides a kind of composite extract, described composite extract is the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei, and described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。At this moment, each effective ingredient being included in described composite extract is described above。
Described test portion can with 1:(0.2～1): (0.2～1): the ratio of (0.2～1) (w/w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
As another embodiment realizing the object of the invention, the present invention provides the compositions of a kind of composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae or its fraction。At this moment, each effective ingredient being included in described composite extract is described above。
The purposes of described compositions can be treatment obesity。
Described composite extract can be that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are with 1:(0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w)。
Described composite extract can pass through to utilize the ethanol (EtOH) of 25% to obtain。
As another embodiment realizing the object of the invention, the present invention provides the compositions of a kind of composite extract containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei or its fraction。At this moment, each effective ingredient being included in described composite extract is described above。
The purposes of described compositions can be treatment obesity。
Described composite extract can be that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are with 1:(0.2～1): (0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w/w)。
Described composite extract can pass through to utilize the ethanol (EtOH) of 25% to obtain。
Beneficial effect
The compositions of the composite extract containing the present invention also is able to hinder lipogenesis under not having Cytotoxic situation such that it is able to be widely applicable for obesity or the prevention of obesity-related disease, improvement or treatment。
It addition, the individual components of the described composite extract such as Radix Polygalae extract shows great toxicity in individuality, hepatic tissue and renal tissue show great chronic inflammatory disease。Therefore, individual components is difficult to individually be administered to individuality。
Accompanying drawing explanation
Fig. 1 a illustrates the inhibition of KIOM-2012Ob (1) hot water extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (1), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (1), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 1 b illustrates the inhibition of KIOM-2012Ob (2) hot water extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (2), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (2), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 1 c illustrates the inhibition of KIOM-2012Ob (3) hot water extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (3), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 1 d illustrates KIOM-2012Ob (1) hot water extract impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (1), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 1 e illustrates KIOM-2012Ob (2) hot water extract impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (2), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 1 f illustrates KIOM-2012Ob (3) hot water extract impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 2 a illustrates the inhibition of KIOM-2012Ob (1) the DW extract (distilled water extraction thing) to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (1), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 2 b illustrates the inhibition of KIOM-2012Ob (2) the DW extract (distilled water extraction thing) to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (2), is combined in the Rhizoma Saururi (Herba Saururi) of KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 2 c illustrates the inhibition of KIOM-2012Ob (3) the DW extract (distilled water extraction thing) to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (3), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 2 d illustrates KIOM-2012Ob (1) the DW extract (distilled water extraction thing) impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (1), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 2 e illustrates KIOM-2012Ob (2) the DW extract (distilled water extraction thing) impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (2), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 2 f illustrates KIOM-2012Ob (3) the DW extract (distilled water extraction thing) impact on the survival rate from the 3T3-L1 cell broken up。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 3 a illustrates the inhibition of KIOM-2012Ob (1) the 25%EtOH extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (3), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (1), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 3 b illustrates the inhibition of KIOM-2012Ob (2) the 25%EtOH extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (3), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (2), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 3 c illustrates the inhibition of KIOM-2012Ob (3) the 25%EtOH extract to the accumulation by oil red O stain neutral fat from the 3T3-L1 adipose cell broken up。Induce differentiation phase to process after KIOM-2012Ob combines crude drug hot water extract 8 days with it from 3T3-L1 and carry out oil red O stain。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob (3), is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。* symbol represents the significance (P < 0.05) that centering athero reduces compared with matched group (0 μ g/ml)。
Fig. 3 d illustrates the impact on the survival rate from the 3T3-L1 cell broken up of KIOM-2012Ob (1) the 25%EtOH extract。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (1), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。$ symbol represents the significance (P < 0.01) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。
Fig. 3 e illustrates the impact on the survival rate from the 3T3-L1 cell broken up of KIOM-2012Ob (1) the 25%EtOH extract。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (2), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。* symbol represents the significance (P < 0.05) that the cells survival rate of survival rate phase comparison KIOM-2012Ob (2) the EtOH extract with Radix Polygalae 25%EtOH extract increases。
Fig. 3 f illustrates the impact on the survival rate from the 3T3-L1 cell broken up of KIOM-2012Ob (3) the 25%EtOH extract。By WST method, the survival rate from the 3T3-L1 cell broken up is measured and represents。Concentration for the treatment of in chart represents with the concentration of KIOM-2012Ob, is combined in the Rhizoma Saururi (Herba Saururi) in KIOM-2012Ob (3), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and processes with identical combined content。$ symbol represents the significance (P < 0.05) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。$ symbol represents the significance (P < 0.01) reduced with matched group (0 μ g/ml) phase comparison cells survival rate。* symbol represents the significance (P < 0.05) that the cells survival rate of cells survival rate phase comparison KIOM-2012Ob (3) the EtOH extract with Radix Polygalae 25%EtOH extract increases。
Fig. 4 illustrates KIOM-2012Ob25%EtOH extract effect of losing weight in fat animal model。Feminine gender (negative) matched group employ the Normal group only supplying 10% food rich in fat, only supplying 60% food rich in fat, the positive (positive) matched group supplying orlistat (Xenical, 62.5mg/kg)+60% food rich in fat。1X in extract-treated groups represents the dosage using one day edible decoction of adult human male as the extract of standard, and 4X represents the 4 times amount administrations with it。The performance of KIOM-2012Ob-4X group is significantly lost weight。
Fig. 5 illustrates the food intake of the mice of supply KIOM-2012Ob25%EtOH extract。* symbol represent the mice of supply KIOM-2012Ob (3) the EtOH extract compared with the food intake of 60%HFD mice food intake reduce significance (P < 0.05)。
Fig. 6 illustrates the KIOM-2012Ob25%EtOH extract inhibition to athero。The negative control group employ the Normal group only supplying 10% food rich in fat, only supplying 60% food rich in fat, the positive controls supplying orlistat (Xenical, 62.5mg/kg)+60% food rich in fat。For the fatty tissue of mice, the weight of abdominal part and subcutaneous fat is measured。* symbol represents the significance (P < 0.05) that the mouse abdominal adipose weight of stomach fat weight phase comparison supply KIOM-2012Ob (3) the EtOH extract with 60%HFD mice reduces。
Fig. 7 illustrates the impact that liver is produced by KIOM-2012Ob25%EtOH extract。Hepatic tissue is fixed on formalin and trust carries out H&E tissue staining and pathology opinion。A represents trickle vesiculovirus lipomatous increase in hepatic tissue, and represents that trickle vesiculovirus lipomatous increase is suppressed in KIOM-2012Ob process group。B represents the increase of chronic inflammatory disease in liver。In KIOM-2012Ob process group, and other extracts from crude drugs process group, especially show the minimizing of significance (P < 0.05, P < 0.01) compared with Radix Polygalae extract process group。$ symbol represents the significance (P < 0.01) of vesiculovirus lipomatous increase fine with Normal group phase comparison。$ symbol represents and the significance (P < 0.05) of the fine vesiculovirus lipomatous increase of general Normal group phase comparison。* symbol represents the significance (P < 0.01) in KIOM-2012Ob process group, chronic inflammatory disease reduced compared with Radix Polygalae extract-4X process group。* symbol represents the significance (P < 0.05) in KIOM-2012Ob process group, chronic inflammatory disease reduced compared with Radix Polygalae extract-4X process group。## symbol represents the significance (P < 0.01) increased with negative control group phase comparison chronic inflammatory disease。# symbol represents the significance (P < 0.05) increased with negative control group phase comparison chronic inflammatory disease。
Fig. 8 illustrates the impact that kidney is produced by KIOM-2012Ob25%EtOH extract。Renal tissue is fixed on formalin and trust carries out H&E tissue staining and pathology opinion。Kidney shows the increase of chronic inflammatory disease。In KIOM-2012Ob process group, compared with other extracts from crude drugs process group, especially show the minimizing of significance (P < 0.05, P < 0.01) compared with Radix Polygalae extract process group。* symbol represents the significance (P < 0.01) in KIOM-2012Ob process group, chronic inflammatory disease reduced compared with Radix Polygalae extract-4X process group。* symbol represents the significance (P < 0.05) in KIOM-2012Ob process group, chronic inflammatory disease reduced compared with Radix Polygalae extract-4X process group。### symbol represents the significance (P < 0.001) increased with negative control group phase comparison chronic inflammatory disease。## symbol represents the significance (P < 0.01) increased with negative control group phase comparison chronic inflammatory disease。# symbol represents the significance (P < 0.05) increased with negative control group phase comparison chronic inflammatory disease。
Fig. 9 is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the body weight of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 10 is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the food intake of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 11 a is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the weight of the abdominal part genitals fat of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 11 b is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the weight of the liver sausage fat of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 11 c is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the weight of the subcutaneous fat of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 11 d for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the chart of the comparative result of total fatty weight of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 12 a is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the level of the Leptin of the activation of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 12 b is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the deacylated Leptin level of the activation of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 12 c is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the Leptin ratio level of the activation of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 13 for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood plasma in the chart of comparative result of leptin level。
Figure 14 a for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of neutral fat level。
Figure 14 b for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of T-CHOL level。
Figure 14 c for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of LDL-cholesterol level。
Figure 14 d for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of HDL-cholesterol level。
Figure 14 e is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of the blood glucose level of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 14 f for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of kreatinin level。
Figure 14 g for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of carbamide level。
Figure 14 h for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of lactic acid dehydrogenase (LDH) level。
Figure 14 i is for illustrating that basis is containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contains the chart of the comparative result of blood alkaline phosphatase (ALP) level of the animal model processing consumption of Rhizoma Acori Graminei or trefoil extract。
Figure 14 j for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of glutamate pyruvate transaminase (GPT) level。
Figure 14 k for illustrating according to containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and optionally contain the process consumption of Rhizoma Acori Graminei or trefoil extract animal model blood in the chart of comparative result of glutamic oxaloacetic transaminase, GOT (GOT) level。
Detailed description of the invention
Below, will the present invention will be described in more detail by embodiment。These embodiments are only intended to the present invention is carried out more specific description, and the present invention is not limited to these embodiments。
Embodiment 1: indivedual extracts of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and compound carry Take the preparation of thing
Each ground product of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei is obtained so that the ratio of table 1 mixes the mixture of 30g, and described mixture is respectively suitable for hot water extraction procedure, warm water extraction method and alcohol extraction procedure, thus obtaining corresponding extract respectively。The described each extract obtained is filtered and obtains liquid phase ingredient, and the liquid phase ingredient obtained is carried out lyophilization and obtains pulverous extract (hot water extract, warm water extraction thing and ethanol extraction), and described pulverous extract is used as so that the concentration of 10mg/ml is dissolved in distilled water the test portion tested subsequently。At this moment, for described hot water extraction, the unboiled water adding 1L in described mixture is heated until 100ml；Described warm water extraction, adds the distilled water of 300ml in described mixture, and extracts 24 hours when maintaining 38 DEG C；For described ethanol extraction, described mixture adds 25% ethanol of 300ml, and extracts 24 hours when maintaining 38 DEG C。
Further, each ground product 30g of described Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei adopt method same as mentioned above prepare indivedual extracts (hot water extract, warm water extraction thing and ethanol extraction) of each composition。
The combination ratio of table 1 extract
Extract Rhizoma Saururi (Herba Saururi) (%) Rhizoma Curcumae Longae (%) Radix Polygalae (%) Rhizoma Acori Graminei (%) Total amount (%)
KIOM-2012Ob(1) 10g(33.3) 10g(33.3) 6g(20) 4g(13.4) 30g(100)
KIOM-2012Ob(2) 10g(33.3) 6g(20) 10g(33.3) 4g(13.4) 30g(100)
KIOM-2012Ob(3) 10g(33.3) 4g(13.4) 10g(33.3) 6g(20) 30g(100)
Embodiment 2: indivedual extracts of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and compound carry Take the In Vitro Anti obesity effect of thing
The 3T3-L1 cell of purchased from American ATCC company is inoculated in containing 10% Ox blood serum (BovineSerum, BS, Gibco) and DMEM (Dulbecco'sModifiedEagle'sMedium, the Lonza) culture medium of 1% Pen .-Strep in, at 37 DEG C, at 5%CO2When cultivate after, and carried out successive transfer culture every 3 to 4 days and obtain Preadipocyte。
The Preadipocyte of described acquisition is reclaimed, and with 1x104After the concentration of cell quantity/ml carries out titration, by its with the amount plant division in 100 μ l/ holes in the microwell plate of 96-hole, at 37 DEG C, at 5%CO2When cultivate 4 days to saturated。Then, described cultured cells is added in described embodiment 1 each Adipocyte Differentiation inducing culture (DMEM containing indivedual extract or composite extract of preparation, 0.25 μM of dexamethasone, 0.5mM3-isobutyl group-1-methylxanthine, 10 μ g/ml insulin and hyclone (FBS, FetalBovineSerum), and further cultivate 2 days。At this moment, the concentration being contained in the described indivedual extracts in Adipocyte Differentiation inducing culture or composite extract is as shown in table 2 below。
The concentration for the treatment of of the other extract of 2, table and composite extract
Then, remove culture medium, and add and cultivate 2 days containing only after having the DMEM culture medium of 10 μ g/ml insulins, changed every two days afterwards once containing only having the DMEM culture medium of FBS and cultivating 8 days。
Reclaim cell after terminating to cultivate, and fix with 10% formalin after washing with PBS。The cell fixed adds oil red O dye, and add 70% isopropanol and wash, cell after dry washing, then 100% isopropanol is added, so that the oil red O on the cell being colored dissolves, and under 490nm, measure absorbance, utilize the absorbance measured to calculate and be deposited in intracellular fat content。At this moment, fat content is represented with the percentage ratio for matched group mean absorbance values。
Additionally, by WST (water solublity tetrazole (Water-solubletetrazolium), 4-[3-(4 iodine substituted phenyl)-2-(4-nitrobenzophenone)-2H-5-tetrazolium]-1,3-benzene disulfonate) (thesodiumsaltof4-[3-(4iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzenedisulfonate) survival rate of the cell of described differentiation is measured by algoscopy, thus evaluating each composite extract and the cytotoxicity of indivedual extract。Roughly, WST solution (10 μ l/100 μ l) is processed to cultivating the cell terminated, thus after cultivating 1 hour further, absorbance (520nm) is utilized to calculate the amount of the first being discharged in culture fluid cured (formazan), thus measuring the cell level of existence, and it is denoted as the ratio of the value for matched group。
First, for the result that the effect suppressing athero of hot water extract is analyzed, as shown in Fig. 1 a to Fig. 1 c, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 and 100 μ g/ml, fat content reduces according to concentration, in indivedual extracts, the fat content that only Radix Polygalae extract performance is similar to composite extract reduces effect, Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract are unrelated with concentration for the treatment of, do not show the effect of different minimizing fat contents。And, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 μ g/ml, fat slip respectively 10%, 13%, 17%；When processing by the concentration of 100 μ g/ml, fat slip respectively 25%, 28% and 26%。
And, hot water extract is carried out the result of cytotoxicity analysis, as shown in Fig. 1 d to 1f, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 and 100 μ g/ml, cell quantity reduces according to concentration, in each indivedual extracts, when processing with Radix Polygalae extract, cell number substantially reduces, when processing with Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract, unrelated with concentration for the treatment of, cell quantity does not reduce。
Then, for the result that the effect suppressing athero of warm water extraction thing is analyzed, as shown in Fig. 2 a to Fig. 2 c, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 and 100 μ g/ml, fat content reduces according to concentration, in each indivedual extracts, the fat content that only Radix Polygalae extract performance is similar to composite extract reduces effect, Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract are unrelated with concentration for the treatment of, do not show the effect of different minimizing fat contents。
And, the result that the thing cytotoxicity of warm water extraction is analyzed, as shown in Fig. 2 d to 2f, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 and 100 μ g/ml, cell quantity reduces according to concentration, in each indivedual extracts, when processing with Radix Polygalae extract, cell number substantially reduces, when processing with Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract, unrelated with concentration for the treatment of, cell quantity does not reduce。
Finally, the result that the effect suppressing athero of ethanol extraction is analyzed, as shown in Fig. 3 a to Fig. 3 c, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 50 and 100 μ g/ml, fat content reduces according to concentration, in each indivedual extracts, the fat content that only Radix Polygalae extract performance is similar to composite extract reduces effect, Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract are unrelated with concentration for the treatment of, do not show the effect of different minimizing fat contents。
And, the result that the thing cytotoxicity of warm water extraction is analyzed, as shown in Fig. 3 d to 3f, to each composite extract (KIOM-2012Ob (1), KIOM-2012Ob (2), KIOM-2012Ob (3)), when processing by the concentration of 100 μ g/ml, cell quantity reduces, in each indivedual extracts, when processing with the Radix Polygalae extract of 50 and 100 μ g/ml concentration, cell number substantially reduces, when processing with Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Rhizoma Acori Graminei extract, unrelated with concentration for the treatment of, cell quantity does not reduce。
The result of described each composite extract (hot water extract, warm water extraction thing and ethanol extraction) is carried out comprehensive after, it is possible to know suppress the effect of athero and cell safety in, the effect of ethanol extraction is the most excellent。
Embodiment 3: indivedual extracts of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei and compound carry Take the internal obesity effect of thing
Whether from the results verification of described embodiment 2, the inhibition of athero and cell safety aspect are shown the effect of excellence by the ethanol extraction of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei in vivo, therefore want to confirm under described ethanol extraction condition in vivo also effective。For this, the indivedual extracts extracted by alcohol extraction procedure and composite extract are ingested to mice and raise, and the body weight of mice raised, food intake and fat weight are measured, and hepatic tissue and renal tissue are compared analysis。
Embodiment 3-1: the acquisition of the mice raised under each different condition
After buying the male C57BL/6N mice 170 of 5 week old and adapting to 1 week, to its 60% food rich in fat 5 weeks raising of ingesting, then often to organize 10, the mice of raising being divided into 17 groups, the feedstuff of the different condition comprising different tests material that it is ingested also raises 6 weeks (referring to table 3) further。At this moment, positive controls is decided to be the group to its known Bariatric agent orlistat of administration。Composite extract is divided into normal diet group (X1) and too much diet group (X4) to be administered。Further, receptacle temperature maintains 22 ± 1 DEG C, and humidity maintains 50 ± 5%, every 12 hours adjusting brightness, 10% food rich in fat and 60% food rich in fat purchased from central laboratory animal company。
The food condition of table 3 mice
Embodiment 3-2: body weight evaluation
Measure the Mouse Weight from the embodiment 3-1 17 kinds of experimental grouies raised and carry out comparative analysis (referring to Fig. 4)。As shown in Figure 4, the body weight confirming experimental group 9 (Radix Polygalae-4 group) reduces with the level most like with Normal group, and the experimental group secondly reducing body weight is experimental group 5 (KIOM-2012Ob (2)-4X group) and 17 (KIOM-2012Ob (3)-4X groups)。The mice of other experimental group still shows the level lower than negative control group, it is known that substantially demonstrate effect of losing weight。
But, for experimental group 9 (Radix Polygalae-4X group), in 6 time-of-weeks, 10 merely hit 4 in the dust, and 1 health worsens seriously, therefore confirm to be subject to toxic insult very serious。In contrast, experimental group 15 (KIOM-2012Ob (2)-4X group) and 17 (KIOM-2012Ob (3)-4X groups) do not demonstrate the infringement being subject to toxicity。
This result is enlightened, although Radix Polygalae extract is effective to losing weight, but owing to showing great toxicity in individuality, it is thus impossible to individually dosed to individuality。
Embodiment 3-3: food intake evaluation
Measure the food intake of the mice from the embodiment 3-1 17 kinds of experimental grouies raised and carry out comparative analysis (referring to Fig. 5)。As it is shown in figure 5, the amount of the food of each experimental group picked-up has some differences, but except experimental group 9 (Radix Polygalae-4X group), it does not have demonstrate significant difference。
Embodiment 3-4: fat weight evaluation
Mice to the 17 kinds of experimental grouies raised from embodiment 3-1, after jejunitas 5 hours and inducing mouse all digest the feedstuff absorbed, and after putting to death above-mentioned mice by the de-bone method of cervical vertebra, takes out stomach fat and subcutaneous fat from each mice。With the stomach fat taken out described in brine and subcutaneous fat, and it is placed in filter paper and gets on moisture removal, then measure the gross weight of these fat and carry out comparative analysis (referring to Fig. 6)。As shown in Figure 6, it is possible to confirm that experimental group 9 (Radix Polygalae-4X group) and experimental group 15 (KIOM-2012Ob (2)-4X group) fat weight significance compared with negative control group reduces。And, to the experimental group 12 (KIOM-2012Ob (1)-1X group) being administered with the composite extract of identical level, experimental group 14 (KIOM-2012Ob (2)-1X group) and experimental group 16 (KIOM-2012Ob (3)-1X group) compare, it is possible to confirm fat sequentially increasing with experimental group 14,16 and 12；Comparative experiments group 13 (KIOM-2012Ob (1)-4X group), experimental group 15 (KIOM-2012Ob (2)-4X group) and experimental group 17 (KIOM-2012Ob (3)-4X group), it is possible to confirm fat sequentially increasing with experimental group 15,17 and 13。Therefore, considering the content of the Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and the Rhizoma Acori Graminei that are included in described each composite extract, it is possible to analyze and be, the fat of described composite extract increases inhibition, and to play the composition of maximum effect be Radix Polygalae, secondly being Rhizoma Curcumae Longae, the inhibition that Rhizoma Acori Graminei plays is minimum。
Embodiment 3-5: the pathological analysis of hepatic tissue
Carry out pathological analysis and the level of the fine lipomatous level of blister type and chronic inflammatory disease that are contained in hepatic tissue is compared analysis (referring to Fig. 7) after taking out hepatic tissue from the mice put to death among described embodiment 3-4 and be fixed on formalin。As shown in Figure 7 A, the fine lipomatous value of blister type during with Radix Polygalae extract (experimental group 8 and 9) with composite extract (experimental group 12-17) is compared with negative control group, all show low value, thus the fine blister type lipoma level significance confirming to be contained in hepatic tissue reduces。
Further, shown in the B in Fig. 7, Radix Polygalae extract (experimental group 8 and 9) greatly brings out chronic inflammatory disease in hepatic tissue, and on the contrary, composite extract (experimental group 12-17) makes these inflammation rates of bringing out substantially reduce。Especially when experimental group 15 and 16, it is shown that with negative control group almost similarity degree bring out rate, thus confirming to bring out chronic inflammatory disease hardly。
These results are enlightened, although Radix Polygalae extract has the fine lipomatous effect of blister type of minimizing, but owing to greatly bringing out chronic inflammatory disease, accordingly, it is difficult to be individually administered to individuality。
In contrast, composite extract not only reduces lipomatous excellent effect, and is in a ratio of very safe material with Radix Polygalae extract。
Embodiment 3-6: the pathological analysis of renal tissue
Carry out pathological analysis and the level of the chronic inflammatory disease being contained in renal tissue is compared analysis (referring to Fig. 8) after taking out renal tissue from the mice put to death among described embodiment 3-4 and be fixed on formalin。As shown in Figure 8, the chronic inflammatory disease level of the renal tissue of all experimental grouies shows high level than negative control group and Normal group, wherein, shows higher level with in the mice of Radix Polygalae extract (experimental group 9)。In contrast, the mice being administered with composite extract (experimental group 12-17) shows the level identical or low with the mice being administered with Radix Polygalae extract。
The result of comprehensive described embodiment 3-2 to 3-6, it is known that Radix Polygalae extract and composite extract show fat inhibition in vivo, although excellent effect fat of suppression, but the Radix Polygalae extract that Animal performance goes out serious toxicity is compared, composite extract shows the safety of excellent fat inhibition and raising。
Embodiment 4: derive from the fat suppression of the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae Effect
From the interpretation of result of described embodiment 3-4 to the composition be contained in described composite extract, Rhizoma Acori Graminei is minimum to fat increase inhibition contribution, therefore, determining and never use Rhizoma Acori Graminei, whether the composite extract that the test portion being only made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae extracts shows fat inhibition。
Embodiment 4-1: the acquisition of Nature inorganic bone thing
Acquisition must comprise the Rhizoma Saururi (Herba Saururi) of pulverizing, Rhizoma Curcumae Longae and Radix Polygalae, and optionally comprises Rhizoma Acori Graminei and trefoil mixture。Add 25% ethanol of 300ml to described mixture, extract 24 hours when maintaining 38 DEG C and obtain each composite extract respectively。Each extract of described acquisition is filtered and obtains liquid phase ingredient, and make the liquid phase ingredient lyophilization of acquisition obtain Powder Extract, and described Powder Extract is dissolved in distilled water with the concentration of 100/ml, thus being used as the test portion tested afterwards。
Further, the green tea extract used in comparative example is also obtained by same method。
The ratio of components of table 4 mixed extract
Embodiment 4-2: the acquisition of the mice raised under each different condition
After buying the male C57BL/6N mice 120 of 5 week old and adapting to 1 week, it is divided into 14 groups often organizing 9, and to the feedstuff of its ingest 60% food rich in fat (HFD) and the different condition that comprises different tests material and raise 8 weeks (referring to table 5) further。At this moment, normal receptacle temperature maintains 22 ± 1 DEG C, and humidity maintains 50 ± 5%, regulates a light and shade every 12 hours, and normal saline solution is used as excipient。
The food condition of table 5 mice
Embodiment 4-3: body weight evaluation
Measure the body weight of the mice of the 14 kinds of experimental grouies raised from described embodiment 4-2 and carried out comparative analysis (referring to Fig. 9)。As shown in Figure 9, the level that the body weight of all experimental grouies (experimental group 27-34) is all low than negative control group performance, the experimental group that body weight reduces with the level most like with Normal group, for experimental group 34 (KSL), is then followed successively by experimental group 28 (K300), experimental group 30 (KS300), experimental group 33 (KCL), experimental group 29 (KS150), experimental group 27 (K150), experimental group 31 (KC150) and experimental group 32 (KC300)。
From described result it is recognised that also show body weight inhibition from the composite extract of the test portion extraction being made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
Embodiment 4-4: food intake evaluation
Measure the food intake of the 14 kinds of experimental mice raised from described embodiment 4-2 and carry out comparative analysis (referring to Figure 10)。As shown in Figure 10, the food intake of each different experiments group has some differences, but does not show the difference of significance。
Embodiment 4-5: fat weight evaluation
After the feedstuff absorbed all is digested by inducing mouse to jejunitas 5 hours of the mice of the 14 kinds of experimental grouies raised from embodiment 4-2, and after putting to death mice by the de-bone method of cervical vertebra, take out fat and subcutaneous fat between abdominal part genitals fat, intestinal from each mice。With the abdominal part genitals fat taken out described in brine, fat and subcutaneous fat be placed on filter paper between intestinal, thus measuring the weight of each fat and gross weight after removing moisture and carrying out comparative analysis (referring to Figure 11 a-to 11d)。
As shown in fig. 11a, the abdominal part genitals fat of all experimental grouies (experimental group 27-34) all shows low level than negative control group, the experimental group that fat weight reduces with the level most like with Normal group, for experimental group 28 (K300), is then followed successively by experimental group 34 (KSL), experimental group 30 (KS300), experimental group 33 (KCL), experimental group 27 (K150), experimental group 32 (KC300), experimental group 29 (KS150) and experimental group 31 (KC150)。
As shown in figure 11b, the level that between the intestinal of all experimental grouies (experimental group 27-34), fat is all low than negative control group performance, the experimental group that fat weight reduces with the level most like with Normal group, for experimental group 30 (KS300), is then followed successively by experimental group 34 (KSL), experimental group 28 (K300), experimental group 33 (KCL), experimental group 32 (KC300), experimental group 29 (KS150), experimental group 27 (K150) and experimental group 31 (KC150)。
As shown in fig. 11c, the level that the subcutaneous fat of all experimental grouies (experimental group 27-34) is all low than negative control group performance, the experimental group that fat weight reduces with the level most like with Normal group, for experimental group 28 (K300), is then followed successively by experimental group 34 (KSL), experimental group 30 (KS300), experimental group 33 (KCL), experimental group 29 (KS150), experimental group 27 (K150), experimental group 32 (KC300) and experimental group 31 (KC150)。
As illustrated in fig. 11d, total fat of all experimental grouies (experimental group 27-34) all shows low level than negative control group, the experimental group that fat weight reduces with the level most like with Normal group is for experimental group 28 (K300), then experimental group 34 (KSL), experimental group 30 (KS300), experimental group 33 (KCL), experimental group 27 (K150), experimental group 29 (KS150), experimental group 32 (KC300) and experimental group 31 (KC150) it are followed successively by。
Comprehensive described Figure 11 a to 11d, it is possible to know that the composite extract performance extracted from the test portion being made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae suppresses the effect of athero。
Embodiment 4-6: the level evaluation of Leptin (ghrelin)
For from stomachial secretion; and it is referred to as the level of the Leptin of hormone (hungerhormone) on an empty stomach; by 14 kinds of experimental mice of raising in described embodiment 4-2 as object, and it is subdivided into the Leptin (activatedghrelin) of activation, deacylated Leptin and Leptin ratio (ghrelinratio) and is evaluated。
First, use Leptin to measure test kit (ActiveGhrelinELISAkit, SCETI, Japan) and measure the level of the Leptin of the activation being present in blood plasma。Generally, the blood obtained from the mice of each experimental group it is placed in the pipe including EDTA and aprotinin and carries out centrifugation and obtain blood plasma after mixing, adding in described blood plasma and be equivalent to the 1mol/LHCl of 10% and prepare test portion。Described ready test portion is added in Leptin mensuration test kit and reacts, under 450nm, measure absorbance, and calculate the level (Figure 12 a) of the Leptin of activation from the absorbance measured。
As figure 12 a shows, Leptin for activation, the experimental group of the level higher than the performance of the value of negative control group is experimental group 32 (KC300), the experimental group showing the value similar to negative control group is experimental group 27 (K150) and experimental group 34 (KSL), the experimental group of the level lower than the performance of the value of negative control group is experimental group 29 (KS150), experimental group 28 (K300), experimental group 33 (KCL), experimental group 30 (KS300) and experimental group 31 (KC150), and the experimental group of performance floor level is experimental group 29 (KS150)。
Then; the level of the deacylated Leptin for existing in blood plasma; except measuring test kit (desacyl-GhrelinELISAkit with deacylated Leptin; SCETI; Japan) replace Leptin to measure test kit (ActiveGhrelinELISAkit; SCETI, Japan), it is measured (referring to Figure 12 b) by above-mentioned identical method。
As shown in Figure 12b; for deacylated Leptin; the experimental group of the level lower than the performance of the value of negative control group is experiment 32 (KC300); the experimental group showing the value similar to negative control group is experimental group 29 (KS150) and experimental group 34 (KSL); the experimental group of the level higher than negative control group performance is experimental group 27 (K150), experimental group 28 (K300), experimental group 33 (KCL), experimental group 30 (KS300) and experimental group 31 (KC150), and the experimental group of performance highest level is experimental group 33 (KCL)。
Finally, Leptin ratio (referring to Figure 12 c) is calculated with the percentage ratio for the level of the Leptin of the activation of the deacylated Leptin of described mensuration。
As shown in fig. 12 c, for Leptin ratio, the experimental group of the level higher than the performance of the value of negative control group is experiment 32 (KC300), the experimental group showing the value similar to negative control group is experimental group 27 (K150), the experimental group of the level lower than negative control group performance is experimental group 34 (KSL), experimental group 29 (KS150), experimental group 28 (K300), experimental group 33 (KCL), experimental group 30 (KS300) and experimental group 31 (KC150), and the experimental group of performance floor level is experimental group 33 (KCL)。
The result of comprehensive described Figure 12 a to 12c; composite extract owing to extracting from the test portion being made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae shows the activation Leptin of similar or low level, deacylated Leptin and Leptin ratio compared with negative control group; as a result it will be appreciated that what the fat inhibition of described composite extract produced not by appetite-suppressing。
Embodiment 4-7: the level evaluation of leptin
, using 14 kinds of experimental mice of raising in described embodiment 4-2 the level of the leptin in blood plasma is evaluated as object。
Specifically used leptin test kit (MouseandRatLeptinELISAkit, mediagnost, Germany) determines the leptin existed in blood plasma。Generally, carry out centrifugation and obtain blood plasma after the blood obtained from the mice of each experimental group being placed in the pipe including EDTA and aprotinin and mixing, add in described blood plasma and be contained in after the dilution buffer of described test kit is diluted, described diluent is joined in above-mentioned leptin test kit and reacts, then measure the absorbance under 450nm, calculate the level (referring to Figure 13) of leptin from the value of the absorbance measured。
As shown in figure 13, leptin level in the blood plasma of all experimental grouies (experimental group 27-34) all shows the level lower than negative control group, the experimental group that leptin in blood plasma reduces with the level most like with Normal group, for experimental group 28 (K300), is then followed successively by experimental group 33 (KCL), experimental group 34 (KSL), experimental group 30 (KS300), experimental group 29 (KS150), experimental group 27 (K150), experimental group 32 (KC300) and experimental group 31 (KC150)。
It is known that, conventionally, be the leptin of the laboratory animal due to inducing obesity opposing type increase, therefore the leptin in blood level increase。Confirm to use the experimental group 28 (K300) of the composite extract from the test portion being made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae provided by the invention extraction to show minimum leptin level, this represents the level that the opposing type that it is leptin is minimum, as a result it will be appreciated that described composite extract shows the effect effectively suppressing obesity to produce。
Embodiment 4-8: the level evaluation of composition in blood
Using 14 kinds of experimental mice raising from described embodiment 4-2 as object, by known method, carry out evaluating (referring to Figure 14 a to 14k) to the level of the composition in the blood such as triglyceride, T-CHOL, LDL-cholesterol, HDL-cholesterol, blood glucose, kreatinin, carbamide, LDH, alkali phosphatase (ALP), glutamate pyruvate transaminase (GPT), glutamic oxaloacetic transaminase, GOT (GOT) in blood。
As shown in figures 14a, for triglyceride (triglyceride), the level that only experimental group 34 (KSL) performance is lower than the value of negative control group, the level that the performance of other experimental group is similar or higher than negative control group to the value of negative control group, the experimental group of performance highest level is experimental group 32 (KC300)。
As shown in fig. 14b, for T-CHOL in blood (totalcholesterol), the experimental group of the level lower than the performance of the value of negative control group is experimental group 27 (K150) and experimental group 34 (KSL), the experimental group showing the value similar to negative control group is experimental group 28 (K300) and experimental group 30 (KS300), the experimental group of the level higher than negative control group performance is experimental group 29 (KS150), experimental group 33 (KCL), experimental group 32 (KC300) and experimental group 31 (KC150), the experimental group of performance highest level is experimental group 33 (KCL)。
As shown in figure 14 c, for LDL-cholesterol in blood, the experimental group of the level higher than the performance of the value of negative control group is experimental group 31 (KC150), experimental group 32 (KC300) and experimental group 33 (KCL), the experimental group showing the value similar to negative control group is experimental group 29 (KS150) and experimental group 30 (KS300), the experimental group of the level lower than the performance of the value of negative control group is experimental group 27 (K150), experimental group 28 (K300) and experimental group 34 (KSL), and the experimental group of performance floor level is experimental group 34 (KSL)。
As shown in Figure 14 d, for HDL-cholesterol in blood, the level that only experimental group 28 (K300) performance is lower than the value of negative control group, the level that the performance of other experimental group is similar or higher than negative control group to the value of negative control group, the experimental group of performance highest level is experimental group 31 (KC150)。
As shown in figure 14e, for blood glucose, the experimental group of the level lower than the performance of the value of negative control group is experimental group 28 (K300) and experimental group 29 (KS150), the experimental group showing the value similar to negative control group is experimental group 34 (KSL), the experimental group of the level higher than the performance of the value of negative control group is experimental group 27 (KC150), experimental group 31 (KC150), experimental group 32 (KC300), experimental group 33 (KCL) and experimental group 30 (KS300), the experimental group of performance highest level is experimental group 32 (KC300)。
As shown in figure 14f, for kreatinin in blood, all experimental grouies (experimental group 27-34) all show the level similar or higher than negative control group to the value of negative control group, the experimental group showing the value similar to negative control group is experimental group 29 (KS150) and experimental group 31 (KC150), the level that other experimental group is all high than negative control group performance, the experimental group of performance highest level is experimental group 34 (KSL)。
As shown in Figure 14 g, for carbamide in blood, all experimental grouies (experimental group 27-34) all show the level higher than negative control group, the experimental group that carbamide increases with highest level is for experimental group 33 (KCL), then experimental group 30 (KS300), experimental group 34 (KSL), experimental group 27 (K150), experimental group 31 (KC150), experimental group 28 (K300), experimental group 29 (KS150) and experimental group 32 (KC300) it are followed successively by。
As shown in Figure 14 h, for lactic acid dehydrogenase in blood (LDH), all experimental grouies (experimental group 27-34) all show the level similar or higher than negative control group to the value of negative control group, the experimental group showing the value similar to negative control group is experimental group 30 (KS300) and experimental group 34 (KSL), the level that other experimental group is all high than the performance of the value of negative control group, the experimental group of performance highest level is experimental group 32 (KC300)。
As shown in Figure 14 i, for blood alkaline phosphatase (ALP), the level that only experimental group 29 (KS150) performance is higher than the value of negative control group, the level that the performance of other experimental group is similar or lower than negative control group to the value of negative control group, the experimental group of performance floor level is experimental group 31 (KC150)。
As shown in Figure 14 j, for glutamate pyruvate transaminase in blood, all experimental grouies (experimental group 27-34) all show the level similar or higher than negative control group to the value of negative control group, the experimental group showing the value similar to negative control group is experimental group 27 (K150), experimental group 28 (K300) and experimental group 30 (KS300), the level that other experimental group is all high than negative control group performance, the experimental group of performance highest level is experimental group 31 (KC150)。
As shown in 14k, for glutamic oxaloacetic transaminase, GOT in blood, all experimental grouies (experimental group 27-34) all show the level similar or higher than negative control group to the value of negative control group, the experimental group showing the value similar to negative control group is experimental group 29 (KS150) and experimental group 34 (KSL), the level that other experimental group is all high than negative control group performance, the experimental group of performance highest level is experimental group 31 (KC150)。
Such as described Figure 14 i to 14k, the blood alkaline phosphatase of experimental group of composite extract that extracts with the test portion being made up of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae provided from the present invention, glutamate pyruvate transaminase, glutamic oxaloacetic transaminase, GOT level and matched group compare, do not show obvious difference, therefore can analyze and not show liver toxicity for described composite extract。

Claims (15)

1. the composite extract of a Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae in preparation for treating the application in the medicine of obesity。
2. application according to claim 1, it is characterised in that described composite extract is that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are with 1:(0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w)。
3. the composite extract of a Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei in preparation for treating the application in the medicine of obesity。
4. application according to claim 3, it is characterized in that, described composite extract is that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are with 1:(0.2～1): (0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w/w)。
5. application according to any one of claim 1 to 4, it is characterised in that described composite extract is by utilizing the ethanol (EtOH) of 25% to obtain。
6. a preparation method for composite extract, described composite extract is the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae, and described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。
7. preparation method according to claim 6, it is characterised in that described test portion is with 1:(0.2～1): the ratio of (0.2～1) (w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
8. a preparation method for composite extract, described composite extract is the composite extract of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei, and described preparation method includes the test portion containing Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei is utilized water, C1～C4Alcohol or their mixed solvent carry out extracting and obtain the step of extract。
9. preparation method according to claim 8, it is characterized in that, described test portion is with 1:(0.2～1): (0.2～1): the ratio of (0.2～1) (w/w/w/w) contains Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
10. a compositions, described compositions contains composite extract or its fraction of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae。
11. compositions according to claim 10, it is characterised in that described composite extract is that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae and Radix Polygalae are with 1:(0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w)。
12. a compositions, described compositions contains composite extract or its fraction of Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei。
13. compositions according to claim 12, it is characterized in that, described composite extract is that Rhizoma Saururi (Herba Saururi), Rhizoma Curcumae Longae, Radix Polygalae and Rhizoma Acori Graminei are with 1:(0.2～1): (0.2～1): the extract of the test portion of the ratio mixing of (0.2～1) (w/w/w/w)。
14. the compositions according to any one of claim 10 to 13, it is characterised in that described compositions is used for Bariatric。
15. the compositions according to any one of claim 10 to 13, it is characterised in that described composite extract is by utilizing the ethanol (EtOH) of 25% to obtain。
CN201410710686.3A 2014-11-28 2014-11-28 A pharmaceutical composition containing a composite extract product of saururus chinensis, turmeric and milkwort root and used for preventing or treating obesity Pending CN105688029A (en)

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Citations (2)

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Publication number Priority date Publication date Assignee Title
US20060286180A1 (en) * 2005-06-16 2006-12-21 Yoko Suetake Lipolysis promoter and food and drink containing the same
KR20130111073A (en) * 2012-03-30 2013-10-10 김철호 Pharmaceutical compositions, food compositions and meat source for preventing or treating obesity related disease comprising extract of lycium chinense, houttuynia cordata and saururus chinensis as active ingredient

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Publication number Priority date Publication date Assignee Title
US20060286180A1 (en) * 2005-06-16 2006-12-21 Yoko Suetake Lipolysis promoter and food and drink containing the same
KR20130111073A (en) * 2012-03-30 2013-10-10 김철호 Pharmaceutical compositions, food compositions and meat source for preventing or treating obesity related disease comprising extract of lycium chinense, houttuynia cordata and saururus chinensis as active ingredient

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