CN103585202A - Plant extract for treating diabetes and preparation method thereof - Google Patents
Plant extract for treating diabetes and preparation method thereof Download PDFInfo
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- CN103585202A CN103585202A CN201210287012.8A CN201210287012A CN103585202A CN 103585202 A CN103585202 A CN 103585202A CN 201210287012 A CN201210287012 A CN 201210287012A CN 103585202 A CN103585202 A CN 103585202A
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Abstract
The invention relates to a plant extract for treating diabetes and a preparation method thereof. The plant extract is from roots of Mirabilis jalapa L., and has effects of lowering blood sugar level, increasing insulin sensitivity and increasing content of skeletal muscle glycogen.
Description
Technical field
The present invention system is about a kind of flower (Mirabilis jalapa L.) root extract of cooking for the treatment of diabetes.
Background technology
Diabetes are chronic diseases that a kind of function defectiveness of pancreas excreting insulin or the insulin effect deficiency of generation cause.Such defect causes the concentration of glucose in blood to increase, thereby causes infringement, particularly blood vessel and the nerve of many tissues or organ.
Diabetes have two kinds of patterns, are called type i diabetes and type ii diabetes.Type i diabetes, claims again insulin dependent diabetes mellitus (IDDM) (insulin-dependent diabetes mellitus, IDDM).IDDM sufferer has the defect of pancreas beta cell, causes normally excreting insulin, and has fatal risk.Conventionally be apt to occur between childhood or juvenile era, thus juvenile onset diabetes be called again, but also gradually find adult sufferer in recent years.Type i diabetes sufferer throughout one's life must be by injection administration insulin, to sustain life.Type ii diabetes, claims again non-insulin-dependent diabetes mellitus (non-insulin-dependent diabetes mellitus, NIDDM), and tool familial, also has close association with envirment factor.Health diet, move and avoid overweight can avoid generation and the deterioration of disease.Under the agitation of exquisite diet, edible too much oils and fats fructose is one of the reason of falling ill of Second-Type diabetes.Type ii diabetes sufferer needs drug administration, to reach desirable glycemic control, also has part sufferer palpus administration of insulin, to reduce blood sugar content.
Diabetes can derive many severe complications, for example, retinopathy, nervous lesion, foot pain, varicosis and swelling, blood circulation imbalance, difficulty in walking, hepatic insufficiency, high-cholesterol disease, albuminuria, hyperlipidemia, prostate enlargement, hypertension etc.Properly control the generation that blood sugar content can effectively be controlled the state of an illness and reduce complication.
The medicine that is used for the treatment of at present diabetes comprises the sulphonyl carbamide Lei, resistance to moral class of MAG (meglitinides), biguanides, thiazolidinediones, and alpha-glucosidase inhibitor, yet it has many restrictions, for example, side effect and a high proportion of Secondary cases drug failure (secondary failure).Therefore, more and more sufferers tend to find the natural antidiabetic medicine of novelty in various medicinal plants.
Cook and spend (Mirabilis jalapa L.), belong to Nyctaginaceae plant, formal name is called Mirabilis jalapa, have another name called that four-o'clock flower, night are tender, night flower, Radix primulae maximowiczii, the flower of having a bath, Rhizoma Anemones flaccidae flower, white lead flower, evening makeup flower, four-o'clock flower, enter DILAOSHU, Flos Hydrangeae paniculatae, snowball, powder tofu pudding, plain perfume etc.Cook flower for herbaceos perennial, and major branch is upright, and side shoot is scattered throws oneself on the ground, and joint portion expands, joint portion aobvious pink, and plant height can reach 1 meter of left and right.Leaf is ovum shape triangle, to life, emerald green, inside has light vein; Flower shape is like loudspeaker, and petal five splits, 2 to 3 centimetres of Hua Jing, and long 4 to 5 centimetres, be mostly bright red or purple, also there are white, powder, Huang and secondary color etc.; Fruit is hard, is black, exactly likes Rhizoma Anemones flaccidae, therefore also claim " Rhizoma Anemones flaccidae flower ".The flower of cooking has been widely used in treating dysentery diuresis, diarrhoea, angina abdominis on Chinese medicine; The diseases such as its root extract also can treat constipation, urinary system disease, acute arthritis.According to plant chemical ingredient analysis, the known colored root of cooking comprises (the C.I.B.Walker such as alkaloid, glycoside, carbohydrate and pohytol, G.Trevisan, M.F.Rossato et al., " Antinociceptive activity of Mirabilis jalapa in mice, " Journal of Ethnopharmacology, vol.120, no.2, pp.169-175,2008.).But there is no document proposes clear and definite experimental data for the extract of the colored root of cooking the effect of its treatment diabetes and the preparation method of distinguishing for separated its effective extract is described.
Summary of the invention
Be different from relevant prior art, the present invention system is about a kind of flower (Mirabilis jalapa L.) root extract of cooking for the treatment of diabetes, this extract be characterised in that with alcohol solution, from the water of the colored root of cooking, slightly extracting thing isolates alcohols insoluble matter or further with this alcohols insoluble matter of desalt, obtain, they are different from the known extracts of cooking, and can be effective to prevention or treatment diabetes.
Therefore, one aspect of the present invention provides a kind of compositions for the treatment of diabetes, it is the insoluble extract of alcohols that comprises flower (Mirabilis jalapa) root of cooking, its HPLC analysis chart that is characterised in that of this extract is within 4.27 and 5.36 minutes, to have absworption peak in the holdup time, and wherein HPLC analysis condition is as follows: instrument: the L-7100 HPLC of Hitachi helps Pu (Hitachi L-7100 HPLC Pump); Volume injected: 50 μ l; Analyze tubing string: Shodex OHpak SB-804HQ, 10 μ m, 8 * 300mm; Exclusion limit (exclusion limit): 1,000,000; Tubing string temperature controller: Eldex CH-150,30 ℃; Mobile phase: water; Flow velocity: 1ml/min; And the loose color reader of evaporative light: Sedere Sedex 75 ELSD, 45 ℃, 3.0 bar.
Specific embodiment according to the present invention, the insoluble extraction system of this alcohols is obtained by the method comprising the following steps: will cook colored root or the dry colored root of cooking, extract to such an extent that slightly extract thing through water; With alcohol solution, in this thick extraction thing, isolate the insoluble extract of an alcohols again.Particularly, the method comprises:
(i) the colored root of cooking of cooking colored root or being dried with water heating, and its filtration is formed to the first filtrate;
(ii) described the first filtrate of concentration step (i) to be to obtain a concentrated filtrate, and is mixed to form a mixed liquor with alcohol solution; And
(iii) the described mixed liquor of filtration step (ii) to be to obtain the second filtrate and an insoluble matter, and collects the insoluble extract of alcohols that this insoluble matter is the colored root of cooking.
On the other hand, the invention provides a kind of compositions for the treatment of diabetes, it is the water insoluble extraction thing that comprises the colored root of cooking, its HPLC analysis chart that is characterised in that of this extract is within 4.85 and 5.58 minutes, to have absworption peak in the holdup time, and wherein HPLC analysis condition is as follows: instrument: the L-7100HPLC of Hitachi helps Pu (Hitachi L-7100 HPLC Pump); Volume injected: 50 μ l; Analyze tubing string: Shodex OHpak SB-804HQ, 10 μ m, 8 * 300mm; Exclusion limit (exclusion limit): 1,000,000; Tubing string temperature controller: Eldex CH-150,30 ℃; Mobile phase: water; Flow velocity: 1ml/min; And the loose color reader of evaporative light: Sedere Sedex 75 ELSD, 45 ℃, 3.0 bar.
Specific embodiment according to the present invention, this water insoluble extraction system is obtained by the method comprising the following steps: will cook colored root or the dry colored root of cooking, extract to such an extent that slightly extract thing through water; With alcohol solution, in this thick extraction thing, isolate the insoluble extract of an alcohols; With water, carry out twice extraction and isolate a water insoluble extraction thing again.Particularly, the method comprises:
(i) the colored root of cooking of cooking colored plant root or being dried with water heating, and its filtration is formed to the first filtrate;
(ii) described the first filtrate of concentration step (i) to be to obtain a concentrated filtrate, and is mixed to form a mixed liquor with alcohol solution;
(iii) the described mixed liquor of filtration step (ii) to be to obtain the second filtrate and an insoluble matter, and collects the insoluble extract of alcohols that this insoluble matter is the colored root of cooking;
(iv) with the described insoluble matter of water dissolution step (iii) centrifugal to collect its precipitate;
(v) again with the described precipitate of water dissolution step (iv) centrifugal to obtain a supernatant; And
(vi) the described supernatant of filtration step (v) to be to obtain the 3rd filtrate and an insoluble matter, and collects this insoluble matter for the water insoluble extraction thing of the colored root of cooking.
For example, alcohol solution of the present invention can be methanol or alcoholic solution, is preferably alcoholic solution.Specific embodiment according to the present invention, concentrated filtrate described herein is to mix with the alcoholic solution of equal-volume.
" ethanol " or " ethanol " that uses herein can be used alternately, and the ethanol representing with percentage ratio or ethanol refer to water-soluble percent by volume (V/V), and for example, 95% ethanol refers to the solution of the water that contains 95% ethanol and 5%.
Again on the one hand, the invention provides the medical composition of a kind of prevention or treatment diabetes, the insoluble extract of alcohols or water insoluble extraction thing that it comprises the colored root of cooking described herein for the treatment of effective dose; An and pharmaceutically acceptable supporting agent.
The present invention also provides a kind of insoluble extract of alcohols of the colored root of cooking described herein or the water insoluble extraction thing purposes for the preparation of the medicine of prevention or treatment diabetes.
Unless otherwise defined, herein the technology of all uses and scientific term with have the knack of in the technical field of the invention this skill person generally understanding person there is identical connotation.
The article that this description is used " one " or " being somebody's turn to do ", unless interior literary composition separately has clear indicating, mean that one or of this article the is above receptor of (that is, at least one).For example, " sample " is to comprise that a plurality of this type of have the knack of sample and the equivalent thereof known to this skill personage in the art.
" diabetes " as used herein second word is a kind of disease of carbohydate metabolism imbalance, comprises " type i diabetes " and " type ii diabetes "." type i diabetes " as used herein is called again " child's type diabetes " or " insulin dependent diabetes mellitus (IDDM) ", it means the disease that pancreas manufacture is few or be feature without insulin, cause is that the function of pancreas manufacture insulin is not good, and sufferer need rely on throughout one's life insulin and control the state of an illness.This patient may cause serious metabolic complication, for example DKA and stupor under without insulin." type ii diabetes " as used herein is called again " adult diabetes mellitus " or " non-insulin-dependent diabetes mellitus ", no matter it means whether have insulin still to produce excessive glucose and to cause circulating glucose too high levels be the disease of feature because bad glucose is cleaned up, the human body that results from is not good to insulin action reaction, but not insulin concentration is not enough.Type ii diabetes is conventionally relevant with diet and obesity.
" extract " as used herein means the product that extracts gained for a material, normally by by the material of wish extraction soak or be mixed in solvent and the solution or the concentrate formulation that obtain.Typically, the preparation of extraction system is from fresh plant or through grinding or dry plant sample.This field is known various extracting process, includes but not limited to dipping, diafiltration, diafiltration again, clears up, counter-current extraction, turbine extraction, extruding/extrusion/squeezing or supercritical fluid carbon dioxide extraction.Suitable solvent includes but not limited to water, ethanol, ethanol/water mixture, methanol, butanols, n-butyl alcohol, isobutanol, acetone, hexane, normal hexane, petroleum ether, ethyl acetate, dichloromethane, chloroform or other solvents.Optionally the concentration of the kind of selective reagent or allotment solvent, extracts to reach suitable polarity.For example, in a preferred embodiment provided by the present invention, be to take water as solvent.Solvent can be about 1:1 to about 1:100(w/v(g/ml with the ratio for the treatment of extracting substance)), (being preferably about 1:1 to about 1:50(w/v(g/ml)), be more preferred from about 1:1 to about 1:20, (w/v(g/ml)), also to be more preferred from about 1:15 or 1:10(w/v(g/ml)).Extraction can be carried out at proper temperature, for example, and at approximately 5 ℃ to approximately 100 ℃, approximately 10 ℃ to approximately 100 ℃, approximately 20 ℃ to approximately 100 ℃, approximately 40 ℃ to approximately 100 ℃, approximately 60 ℃ to approximately 100 ℃, approximately 70 ℃ to approximately 100 ℃ or approximately 80 ℃ to approximately 100 ℃.In one example, the root of fresh plant or dry plant sample (optionally through chopping or grind) is mixed with water or soaks also abundant stirring and reach one section of time enough, for example, 1 hour or above, 4 hours or above, 8 hours or above, 10 hours or above, 12 hours or above, 14 hours or above 16 hours or more than, in room temperature, carry out or heat and carry out, via filtration, remove solid residue (filtering residue), then collect the juice (slightly extracting liquid) obtaining; Concentrated thick extraction liquid optionally, and add other solvents further to be cut apart or separated as alcohol solution.Gained is cut apart or the extract of separation also can add solvent further to give purification as water again.
" treatment " as used herein second word comprises the deformity that symptom, this disease in order to cure, heal, alleviate, releive, change, correct, improve, improve or affect this disease, this disease causes or the object of suffering from the tendency of this disease, and the compositions that comprises one or more activating agent is used or administration to suffering from the symptom of this disease, this disease or having the individuality of the tendency of suffering from this disease, or these individualities are carried out to other processing.For example, treatment diabetes generally include bestows sufferer one active component or medicine, reduces blood glucose concentration, improves insulin sensitivity, the object of the consumption of reduction insulin resistance or increase glucose etc. to reach." prevention " second diction that this description is used means as individual premorbid anticipates and can avoid producing this disease.
" individuality " as used herein second word refers in particular to the mammal that needs treatment described herein, for example, the mankind, but also can be house pet (as dog, cat etc.), farming animals animal (as cattle, sheep, horse, pig etc.) or laboratory animal (as, white mice, rat sky, Zhu Mus, white rabbit etc.).
" treatment effective dose " second word that this description is used means that the material being used for the treatment of can provide the amount of therapeutic efficiency, it is to depend on the pattern used and situation to be treated, includes but not limited to age, body weight, symptom, therapeutic effect, dispensing path and treatment time.
" supporting agent " that this description is used or " pharmaceutically acceptable supporting agent " second word refer to diluent, excipient or the analog for medicine, comprise the pharmacy industry general knowledge person well known master of institute.
Can use the present invention's medical composition in any applicable path, include but not limited to parenteral or oral administration medicine supplying.Its form of the medical composition of parenteral dispensing comprises solution, suspension, emulsion, and can carve before use dissolving or be suspended in the solid Injectable composition in solvent.Can in diluent, prepare this injection by dissolving, suspension or emulsifying one or many active component.The example of aforementioned diluent is distilled water, normal saline, vegetable oil, alcohols and the combination thereof for injection.Again, this injection can contain tranquilizer, cosolvent, suspending agent, emulsifying agent, smooth agent, buffer, preservative agent etc.These injection tie up in final preparation step sterilizing or prepare with sterile procedure.The present invention's medical composition also can be formulated to sterile solid preparation, for example, by lyophilization, and can carve before use sterilizing or be dissolved in sterile injectable water or other sterile diluents.
According to the present invention, this medical composition is preferably oral administration, and wherein said composition can be solid or liquid form.Solid composite comprises lozenge, pill, capsule, dispersibility powder, granule and analog thereof.Orally administered composition also comprises mouth-wash and sublingual tablet.Capsule comprises hard capsule and soft capsule.In this type of Orally-administered solid composition, one or many reactive compound can mix voluntarily, or mixes with diluent, bonding agent, the agent of loose solution, lubricant, tranquilizer, cosolvent, then with prior art method preparation, becomes preparation.When needed, these preparations can smears coating, or can two or the coating of multiple coating layer.On the other hand, liquid oral compositions comprises pharmaceutically acceptable liquid solution, suspension, emulsion, syrup, medicated wine, and analog.In such composition, one or many reactive compound can be dissolved, suspend or be emulsified in all-purpose diluent (as mixture of purified water, ethanol or its etc. etc.).Except this type of diluent, foregoing also can contain wetting agent, suspending agent, emulsifying agent, sweeting agent, flavoring agent, spice, preservative agent and buffer and analog thereof.
In following specific embodiment, the present invention will more specifically be described.Specific embodiment that it should be noted that what follows the present invention is only for explanation, is not the particular form being listed as in detail or the limit scope of the invention is extremely disclosed.
Accompanying drawing explanation
For explaination object of the present invention, in the current preferred embodiment of graphic middle demonstration.Yet, it should be understood that the preferred embodiment the invention is not restricted in graphic.
Fig. 1 shows the preparation flow schematic diagram of the insoluble extract of alcohols of the colored root of cooking of the present invention.
Fig. 2 shows the preparation flow schematic diagram of the water insoluble extraction thing of the colored root of cooking of the present invention.
Fig. 3 provides the HPLC analysis chart of Pu Lulan (Pullulan) polysaccharide standard substance P-800, P-200, P-50 and P-10 aqueous solution mixed in equal amounts.
Fig. 4 provides the HPLC analysis chart of Pu Lulan (Pullulan) polysaccharide standard substance P-400, P-100, P-20, P-5 aqueous solution mixed in equal amounts.
Fig. 5 provides the HPLC analysis chart of the present invention's alcohols insoluble (50%EtOH-insol) extract.
Fig. 6 provides the HPLC analysis chart of the present invention's alcohols solubility (50%EtOH-sol) extract.
Fig. 7 provides the HPLC analysis chart of the present invention's water-insoluble (H2O-insol) extract.
Fig. 8 A, Fig. 8 B and Fig. 8 C show that the present invention's the insoluble extract of alcohols (50%EtOH-insol) is for the impact of the glucose tolerance of Second-Type diabetic mice, and wherein Fig. 8 A to C represents that respectively mice is in the variation situation of each time point blood glucose value of lumbar injection glucose front and back for the treatment of the 0th week, the 1st week and the 2nd week.
Fig. 9 shows that the insoluble extract of the present invention's alcohols (50%EtOH-insol) is for the impact of the glycogen content of the skeletal muscle of Second-Type diabetic mice, wherein "
*" show that the present invention's alcohols effect that insoluble extract is given birth to and negative matched group compare, have to add up and go up remarkable difference (p value <0.05).
Figure 10 A and Figure 10 B show that the present invention's the insoluble extract of alcohols (50%EtOH-insol) is for the impact of the glucose tolerance of Second-Type diabetic mice, wherein Figure 10 A provide per os feed with glucose before and after the variation situation of each time point blood glucose value; And Figure 10 B provides the statistical result (AUC: area under curve) of glucose in blood utilization rate.
Figure 11 A and Figure 11 B show that the present invention's the insoluble extract of alcohols (50%EtOH-insol) is for the impact of the sucrose toleration of Second-Type diabetic mice, and wherein Figure 11 A represents that per os is fed and the variation situation of each time point blood glucose value of sucrose front and back; And Figure 11 B provides the statistical result (AUC: area under curve) of glucose in blood utilization rate.
Figure 12 A and Figure 12 B show that the present invention's alcohols solubility extract (50%EtOH-sol) is for the impact of the intravenous glucose injection toleration of Second-Type diabetic mice, and wherein Figure 12 A and Figure 12 B are shown in respectively the 7th and the variation situation of 14 days measured each time point blood glucose values.
The specific embodiment
(1) prepare example
Prepare the insoluble extract of example 1(ethanol)
Get the colored root dried powder of cooking of requirement, add 10 times of volume water of about dried powder gross weight to extract; Filtration gained filtrate is evaporated to approximately 1/3 of remaining former filtrate volume, add again with 95% alcoholic solution of this concentrated filtrate equal-volume and extract for the second time, retaining after filtering filtrate and be ethanol solubility extract and collecting insoluble matter is the insoluble extract of ethanol, prepares to carry out high-performance liquid chromatograph (High-Performance Lipid Chromatography; HPLC) analyze.Fig. 1 is the preparation flow schematic diagram of the insoluble extract of ethanol.
Prepare example 2(water insoluble extraction thing)
According to the extraction step of preparing example 1, extract to obtain the insoluble extract of ethanol, add suitable quantity of water to this insoluble matter to extract for the third time, and centrifuging and taking obtains after its precipitate, add again the water of approximately 40 to 50 times of volumes of this precipitate to carry out the 4th thing extraction, and centrifugal to obtain its supernatant and to filter, collecting after filtering insoluble matter is water insoluble extraction thing, prepares to carry out HPLC analysis.Fig. 2 is the preparation flow schematic diagram of water insoluble extraction thing.
HPLC analyzes
With molecular screening chromatography (size exclusion chromatography) analytic sample, and use gel permeation chromatography (gel filtration chromatography) tubing string, composition is sequentially rushed and carried from large to small according to molecular weight.Compare the holdup time (RT) with standard substance, in order to estimate molecular weight (MW) simultaneously.
I. the preparation of standard solution: 8 Pu Lulan (pullulan) polysaccharide standard substance (P-5 ,-10 ,-20 ,-50 ,-100 ,-200 ,-400 ,-800) are configured to respectively to 1mg/mL aqueous solution, add the water mitigation rearmounted cold preservation of stirring (4 ℃) overnight, to guarantee being dissolved into aqueous solution after its imbibition.To analyze for HPLC after 0.45 μ m membrane filtration.
II. the preparation of extract solution: testing sample is respectively mixed with to 3mg/mL aqueous solution with water, insoluble person optionally after heated and stirred (30 minutes, bubble) under room temperature, cooling and cold preservation (4 ℃) is overnight.To analyze for HPLC after 0.45 μ m membrane filtration.
III.HPLC condition:
Instrument: the L-7100HPLC of Hitachi helps Pu (Hitachi L-7100 HPLC Pump); Volume injected: 50 μ l; Analyze tubing string: Shodex OHpak SB-804HQ, 10 μ m, 8 * 300mm; Exclusion limit (exclusion limit): 1,000,000; Tubing string temperature controller: Eldex CH-150,30 ℃; Mobile phase: water; Flow velocity: 1mL/min; And the loose color reader of evaporative light: Sedere Sedex 75 ELSD, 45 ℃, 3.0 bar.
VI. analysis result:
(i) standard substance
As shown in Figure 3, the HPLC analysis chart that P-400, P-100, P-20 and P-5 mix as shown in Figure 4 for the HPLC analysis chart of standard substance P-800, P-200, P-50 and P-10 aqueous solution mixed in equal amounts.Molecular weight and the holdup time thereof of each standard substance correspondence are shown in table 1:
Table 1
| Standard substance | Mp (the molecule weight of crest) | RT (holdup time, minute) |
| P-800 | 708,000 | 5.4 |
| P-400 | 344,000 | 5.8 |
| P-200 | 200,000 | 6.4 |
| P-100 | 107,000 | 6.9 |
| P-50 | 47,100 | 7.6 |
| P-20 | 21,100 | 8.1 |
| P-10 | 9,600 | 8.6 |
| P-5 | 5,900 | 8.8 |
(ii) testing sample
The HPLC analysis chart of the aqueous sample of ethanol described herein insoluble (50%EtOH-insol), ethanol solubility (50%EtOH-sol) and water-insoluble (H2O-insol) extract is respectively as shown in Fig. 5-7.Compared to standard substance, each testing sample ingredient molecular weight distribution and relative amount are shown in table 2.As shown in Fig. 5-7, the insoluble extract of ethanol is within 4.27 and 5.36 minutes, to have 2 main crests in the holdup time, in the holdup time, is within 6.33 and 9.94 minutes, to have 2 less important crests; Ethanol solubility extract is within 9.93 minutes, to have 1 main crest in the holdup time, in the holdup time, is within 6.49,6.73,7.13,7.29 and 7.46 minutes, to have 5 less important crests; Water insoluble extraction thing had 2 main crests in 4.85 and 5.58 minutes, in the holdup time, be within 5.93 and 6.97 minutes, to have 2 less important crests.
Table 2
(2) effect example
Example 1: the insoluble extract of ethanol (50%EtOH-insol) is for the impact of the lumbar injection glucose tolerance of Second-Type diabetic mice
ICR mice (approximately 4 week age) first fasting before experiment with the Second-Type diabetes that high fat and high fructose diet brought out is overnight.Mice is divided into three groups of dispensing experiments of carrying out 15 days by a definite date: negative matched group, 50%EtOH-insol group and positive control group.Anaesthetize and take a blood sample (0min) every day with pentobarbital (pentobarbital), the person of connecing respectively the metformin (Metformin) of oral administration ddH2O to the insoluble extract of ethanol of negative matched group, 80mg/kg to 50%EtOH-insol group and 300mg/kg to positive control group.Then, then with the glucose of lumbar injection 2g/kg to each mice.After starting dispensing the 1st, 8 and 15 days, take a blood sample for 30,60,120 and 150 minutes after injectable dextrose monohydrate, to measure the content of blood glucose and insulin in serum, and record Mouse Weight simultaneously.As shown in Fig. 8 A to C, after treatment one week, with respect to negative matched group, the insoluble extract of ethanol has obviously improved 30 and the blood glucose value of 60min; After two weeks, significantly reduce by 120 and the blood glucose value of 150min.The insoluble extract of ethanol also can improve the fasting insulin content (table 3) for the treatment of after two weeks.In addition, as shown in table 4 and table 5, the insoluble extract of ethanol not only significantly reduces ingesting and amount of drinking water after treatment, also can alleviate the rear body weight of mice treatment.Again, after 15 days, mice is sacrificed, and take out skeletal muscle and assess its glycogen synthetic quantity, also find that the present invention's the insoluble extract of ethanol can increase the glycogen content (Fig. 9) of mice skeletal.
Table 3
Result is to represent with meansigma methods ± SEM, "
*" show the p value <0.05 compared to negative matched group.
Table 4
Result is to represent with meansigma methods ± SEM, "
*" show the p value <0.05 compared to negative matched group; "
* *" show the p value <0.005 compared to negative matched group.
Table 5
Result is to represent with meansigma methods ± SEM, "
*" show the p value <0.05 compared to negative matched group.Example 2: the insoluble extract of ethanol (50%EtOH-insol) is for the impact of the oral glucose tolerance of Second-Type diabetic mice
ICR mice (approximately 4 week age) first fasting before experiment with the Second-Type diabetes that high fat and high fructose diet brought out is overnight.Mice is divided into three groups: negative matched group, 50%EtOH-insol group and positive control group.Before experiment, 30min first distinguishes oral feeding and ddH
2the metformin (Metformin) of O to the insoluble extract of ethanol of negative matched group, 80mg/kg to 50%EtOH-insol group and 300mg/kg is to positive control group.By take a blood sample after mouse anesthesia (0min), the glucose of each mice 1g/kg of the oral administration of the person of connecing.Extract the blood of the 15th, 30,60,120 and 150min after administration glucose, to measure the content of blood glucose and insulin in serum.As shown in Figure 10 A to B, with respect to negative matched group, the insoluble extract of ethanol obviously improves the blood glucose value of each time point.Further detect again serum insulin content, find that the present invention's the insoluble extract of ethanol improves glucose utilization rate, not see through and affect (table 6) due to amount of insulin secretion.
Table 6
Result system represents with meansigma methods ± SEM.
Example 3: the insoluble extract of ethanol (50%EtOH-insol) is for the impact of the oral sucrose toleration of Second-Type diabetic mice
ICR mice (approximately 4 week age) first fasting before experiment with the Second-Type diabetic mice that high fat and high fructose diet brought out is overnight.Mice is divided into five groups: negative matched group, GYT20, GYT40, GYT80 and positive control group.Before experiment, 30min first distinguishes oral feeding and ddH
2the acarbose (Acarbose) of the ethanol insoluble extract of O to the insoluble extract of ethanol of negative matched group, 20mg/kg to GYT20,40mg/kg to the insoluble extract of ethanol of GYT40,80mg/kg to GYT80 and 10mg/kg is to positive control group.By take a blood sample after mouse anesthesia (0min), the sucrose of each mice 2g/kg of the oral administration of the person of connecing.Extract the blood of the 15th, 30,60,120 and 150min after administration sucrose, to measure the content of blood glucose and insulin in serum.As shown in Figure 11 A to B, with respect to negative matched group, the obvious improvement 30 of the insoluble extract of ethanol of 20mg/kg and the blood glucose value of 60min, and the insoluble extract of the ethanol of 40mg/kg and 80mg/kg has similar effect of lowering blood sugar to positive control group, it all can improve the blood glucose value of each time point.Further detect again the content of serum insulin, find that the present invention's the insoluble extract of ethanol improves sucrose utilization rate, not see through and affect (table 7) due to amount of insulin secretion.
Table 7
Result system represents with meansigma methods ± SEM.
Example 4: ethanol solubility extract (50%EtOH-sol) is for the impact of the intravenous glucose injection toleration of Second-Type diabetes rat
The Second-Type diabetes Wistar rat of being brought out with streptozotocin (streptozotocin) and nicotiamide (nicotinamide) (approximately 14 week age) first fasting 16 hours before experiment, with isoflurane (Isoflurane) anesthesia and from tail vein blood, time point is labeled as-30 again.Rat is divided into matched group and 50%EtOH-sol group to carry out the dispensing experiment of 14 days by a definite date.By rat anesthesia oral feeding with ddH2O to the ethanol solubility extract of matched group and 48mg/kg to 50%EtOH-sol group respectively after 30 minutes, the person of connecing is from the glucose of tail vein injection 1g/kg.After starting dispensing the 7th and 14 days, 30,60,90 and 120min after injectable dextrose monohydrate take a blood sample, to measure the content of blood glucose and insulin in serum.As shown in Figure 12 A to B, 50%EtOH-sol is not had an effect of blood sugar lowering with respect to matched group.
Claims (14)
1. a compositions for the treatment of diabetes, it is the insoluble extract of alcohols that comprises flower (the Mirabilis jalapa L.) root of cooking, this extract is characterised in that its HPLC analysis chart is within 4.27 and 5.36 minutes, to have absworption peak in the holdup time, and wherein HPLC analysis condition is as follows: instrument: the L-7100 HPLC of Hitachi helps Pu (Hitachi L-7100 HPLC Pump); Volume injected: 50 μ l; Analyze tubing string: Shodex OHpak SB-804HQ, 10 μ m, 8 * 300mm; Exclusion limit (exclusion limit): 1,000,000; Tubing string temperature controller: Eldex CH-150,30 ℃; Mobile phase: water; Flow velocity: 1ml/min; And the loose color reader of evaporative light: Sedere Sedex 75 ELSD, 45 ℃, 3.0 bar.
2. according to the compositions of claim 1, wherein the insoluble extraction system of this alcohols is obtained by the method comprising the following steps:
By colored root or the dry colored root of cooking of cooking, through water, extract to such an extent that slightly extract thing; With alcohol solution, from this thick extraction thing, isolate the insoluble extract of an alcohols again.
3. according to the insoluble extract of the alcohols of claim 2, it is obtained by the method comprising the following steps:
(i) the colored root of cooking of cooking colored root or being dried with water heating, and its filtration is formed to the first filtrate;
(ii) described the first filtrate of concentration step (i) to be to obtain a concentrated filtrate, and is mixed to form a mixed liquor with alcohol solution; And
(iii) the described mixed liquor of filtration step (ii) to be to obtain the second filtrate and an insoluble matter, and to collect this insoluble matter be the insoluble extract of alcohols of colored root of cooking.
4. according to the insoluble extract of the alcohols of claim 3, wherein this concentrated filtrate system is mixed with 95% (v/v) above alcohol solution of equal-volume.
5. according to the insoluble extract of the alcohols of claim 4, wherein this alcohol solution is methanol or ethanol.
6. according to the insoluble extract of the alcohols of claim 5, wherein this alcohol solution is ethanol.
7. a compositions for the treatment of diabetes, it is the water insoluble extraction thing that comprises the colored root of cooking, this extract is characterised in that its HPLC analysis chart is within 4.85 and 5.58 minutes, to have absworption peak in the holdup time, and wherein HPLC analysis condition is as follows: instrument: the L-7100HPLC of Hitachi helps Pu (Hitachi L-7100 HPLC Pump); Volume injected: 50 μ l; Analyze tubing string: Shodex OHpak SB-804HQ, 10 μ m, 8 * 300mm; Exclusion limit (exclusion limit): 1,000,000; Tubing string temperature controller: Eldex CH-150,30 ℃; Mobile phase: water; Flow velocity: 1ml/min; And the loose color reader of evaporative light: Sedere Sedex 75 ELSD, 45 ℃, 3.0 bar.
8. according to the compositions of claim 7, wherein this water insoluble extraction system is obtained by the method comprising the following steps:
By colored root or the dry colored root of cooking of cooking, through water, extract to such an extent that slightly extract thing; With alcohol solution, from this thick extraction thing, isolate the insoluble extract of an alcohols; With water, carry out twice extraction and isolate a water insoluble extraction thing again.
9. water insoluble extraction thing according to Claim 8, it is obtained by the method comprising the following steps:
(i) the colored root of cooking of cooking colored root or being dried with water heating, and its filtration is formed to the first filtrate;
(ii) described the first filtrate of concentration step (i) to be to obtain a concentrated filtrate, and is mixed to form a mixed liquor with alcohol solution;
(iii) the described mixed liquor of filtration step (ii) to be to obtain the second filtrate and an insoluble matter, and to collect this insoluble matter be the insoluble extract of alcohols of colored root of cooking;
(iv) with the described insoluble matter of water dissolution step (iii) centrifugal to collect its precipitate;
(v) again with the described precipitate of water dissolution step (iv) centrifugal to obtain a supernatant; And
(vi) the described supernatant of filtration step (v) to be to obtain the 3rd filtrate and an insoluble matter, and to collect this insoluble matter be the water insoluble extraction thing of colored root of cooking.
10. according to the water insoluble extraction thing of claim 9, wherein this concentrated filtrate system is mixed with 95% (v/v) above alcohol solution of equal-volume.
11. according to the water insoluble extraction thing of claim 10, and wherein this alcohol solution is methanol or ethanol.
12. according to the water insoluble extraction thing of claim 11, and wherein this alcohol solution is ethanol.
13. 1 kinds of medical compositions that prevent or treat diabetes, it comprises treats the insoluble extract of the alcohols according to claim 1 of effective dose or the water insoluble extraction thing of claim 7; An and pharmaceutically acceptable supporting agent.
The insoluble extract of alcohols of 14. 1 kinds of claim 1 or the water insoluble extraction thing of claim 7 are for the preparation of the purposes of the medicine of prevention or treatment diabetes.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210287012.8A CN103585202A (en) | 2012-08-13 | 2012-08-13 | Plant extract for treating diabetes and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210287012.8A CN103585202A (en) | 2012-08-13 | 2012-08-13 | Plant extract for treating diabetes and preparation method thereof |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225082A (en) * | 2011-06-22 | 2011-10-26 | 中国人民解放军第三军医大学第二附属医院 | Medicament for preventing and treating diabetes and complications thereof and preparation method thereof |
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- 2012-08-13 CN CN201210287012.8A patent/CN103585202A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102225082A (en) * | 2011-06-22 | 2011-10-26 | 中国人民解放军第三军医大学第二附属医院 | Medicament for preventing and treating diabetes and complications thereof and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 刘安军等: "紫茉莉根水提物对葡萄糖及四氧嘧啶致高血糖小鼠的降血糖作用", 《现代食品科技》, vol. 27, no. 2, 15 February 2011 (2011-02-15) * |
| 周吉银等: "紫茉莉提取方法、化学成分及药理作用研究概述", 《中国医院药学杂志》, vol. 31, no. 16, 30 August 2011 (2011-08-30), pages 1376 - 1379 * |
| 李娟好等: "紫茉莉根水提物降血糖作用的研究", 《广东药学院学报》, vol. 22, no. 3, 25 June 2006 (2006-06-25), pages 299 - 305 * |
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