CN101239091A - Composition for preventing and controlling helicobacter pylori affection - Google Patents
Composition for preventing and controlling helicobacter pylori affection Download PDFInfo
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- CN101239091A CN101239091A CNA2007100372466A CN200710037246A CN101239091A CN 101239091 A CN101239091 A CN 101239091A CN A2007100372466 A CNA2007100372466 A CN A2007100372466A CN 200710037246 A CN200710037246 A CN 200710037246A CN 101239091 A CN101239091 A CN 101239091A
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Abstract
The invention provides a composition which includes: procyanidins, fuling polysaccharide and dandelion, and is also related to the application of the composition in preparing drug for curing and/or preventing helicobacter pylori-related infectious diseases, and food or health products.
Description
Technical field
The present invention relates to drug world.More particularly, the present invention relates to a kind of compositions that can prevent and treat the helicobacter pylori infections effect, and said composition is used for preventing and treating the purposes of the medicine of the disease relevant with helicobacter pylori infections in preparation.
Background technology
Nineteen eighty-three Warren and Marshall have found helicobacter pylori (Helicobacter pylori).Now confirm, helicobacter pylori be the main diseases that causes gastritis (as chronic superficial gastritis, chronic atrophic gastritis etc.), peptic ulcer (as duodenal ulcer and gastric ulcer) because of, also closely related with the generation of gastric cancer.It is believed that the individuality generation gastric tumor that infects helicobacter pylori with MALT (the relevant lymph sample of mucosa) the high 2-8 of lymphadenomatous risk times.World Health Organization (WHO) in last century the nineties classify helicobacter pylori as I level carcinogen.The elimination of helicobacter pylori all has important meaning for prevention, treatment and the prognosis of above-mentioned disease.Therefore, to the treatment of chronic gastropathy, except that using traditional medicine, also should give the anti helicobacter pylori medicine thing.Therefore Warren in 2005 and Marshall have obtained Nobel's medical science or physiology prize.
China adult helicobacter pylori infections rate is up to more than 60%.Helicobacter pylori can enter gastric by contaminant water and food after excrement-mouth or the infection of mouth-mouth approach.Gastric helicobacter pylori major part is free in the slime layer, has only minority to be incorporated into gastric epithelial cell by its excretory adhesin.The adhesion of helicobacter pylori is its essential condition that is colonizated in gastric epithelial.Helicobacter pylori and gastric epithelial cell are combined closely, and cause a series of downstream events, cause gastric mucosal lesion.Therefore by suppressing or blocking helicobacter pylori and sticking of gastric epithelial cell can prevent and treat helicobacter pylori infections.Simultaneously, discover that the part patient only produces one and crosses impatient sexuality and dye and removed naturally by body, the power that the infected's autospecific immunne response is described also is vital for the removing of helicobacter pylori infections.
Helicobacter pylori eradication method commonly used clinically at present mainly is a Drug therapy, adds two kinds of antibacterials by antacid and forms.Mechanism of action is the bactericidal action that utilizes antibacterials, by suppressing the synthetic mechanism that waits of bacterioprotein with killing bacteria.Though these Drug therapys can obtain certain curative effect, but the gastritis that can not prevent helicobacter pylori infections and cause, and also have very big drawback in the treatment, as drug side effect is big, patient's compliance is poor, particularly Resistant strain appearance and the recurrence of treatment back etc.Especially what deserves to be explained is, generally use or even abuse along with antibacterials, the incidence rate of former or secondary drug resistance helicobacter pylorus bacteria strain rises just year by year, for example the Shanghai City helicobacter pylori to the resistant rate of metronidazole by 30% of nineteen ninety-five rise to 1999 70%.The appearance of Resistant strain at present is one of clinical eradicate helicobacter pylori failure most important reason.
There is the natural Chinese herbal medicines resource in China, studies show that many Chinese herbal medicine all have direct repression to helicobacter pylori, and present anti-helicobacter pylori Chinese medicine preparation at this mechanism of action exploitation, does not still obtain the ideal preparation of curative effect all the time yet.
Therefore, still need to develop the anti-helicobacter pylori Chinese medicine preparation that makes new advances in this area.
Summary of the invention
Natural plants, the especially Chinese herbal medicine that exploitation has the different mechanisms of action or its extract will be the directions of following anti-helicobacter pylori research.The advantage of this class natural plants or its extract is that 1. side effect are little; 2. can not produce drug resistance.The mechanism of natural plant extracts can be sticking and regulate immunity of organisms etc. between blocking-up helicobacter pylori and the gastric epithelial cell.The present inventor adds pachyman and these two kinds Chinese medicine or the extracts that can regulate the body cell immunity of Herba Taraxaci in the procyanidin (or cranberry fruit juice) to by discovering, can improve the preventive and therapeutic effect to helicobacter pylori significantly.
For achieving the above object, first aspect present invention provides a kind of compositions, it is characterized in that, described compositions comprises procyanidin, pachyman and the Herba Taraxaci that treats and/or prevents the helicobacter pylori infections effective dose.
At one more preferably in the embodiment, described compositions also comprises pharmaceutically acceptable carrier.
At another more preferably in the embodiment, in described compositions, the content of described procyanidin is the 0.02-100 weight portion, the content of described pachyman is the 0.1-100 weight portion, the content of described Herba Taraxaci is the 1-100 weight portion, and the gross weight of described procyanidin, pachyman and Herba Taraxaci is the 0.01-99.9% weight of composition weight.
At one more preferably in the embodiment, described compositions comprises cranberry or its extract, pachyman and the Herba Taraxaci that treats and/or prevents the helicobacter pylori infections effective dose.Described cranberry or its extract are preferably cranberry fruit juice.Especially, the ratio of each component is in the described compositions, contains 0.1-100 gram pachyman and 1-100 gram Herba Taraxaci in every liter of cranberry fruit juice.
Compositions of the present invention has any dosage form of acceptable on bromatology or the pharmaceutics, for example comprises oral liquid, tablet, capsule, electuary, granule, pill, powder, teabag, beverage class or injection.
The present invention relate on the other hand compositions of the present invention preparation be used for the treatment of and/or the medicine of the disease that prevention is relevant with helicobacter pylori infections in application.
The present invention relates to the application of compositions of the present invention in preparation food or health product on the other hand.
The present inventor combines procyanidin and natural Chinese herbal medicines (pachyman and Herba Taraxaci) or its extract, has obtained to treat and/or prevent significantly the effect of helicobacter pylori infections.And compositions of the present invention is the Chinese herbal medicine of natural plant extracts and integration of edible and medicinal herbs, therefore also has advantages such as mechanism of action uniqueness, safety, good effect.
Description of drawings
Fig. 1 shown under 750 power microscopes observed behind helicobacter pylori infections the matched group gastric mucosa Giemsa dyeing situation in 8 weeks.
Fig. 2 has shown at observed helicobacter pylori infections under 750 power microscopes after the gastric mucosa Giemsa in 4 weeks dyeing situation behind cranberry fruit juice, pachyman and the Herba Taraxaci combined therapy.
Specific embodiments
Particularly, first aspect present invention provides a kind of compositions, it is characterized in that, described compositions comprises procyanidin, pachyman and the Herba Taraxaci that treats and/or prevents the helicobacter pylori infections effective dose.
Be applicable to that procyanidin of the present invention, pachyman and Herba Taraxaci all are well known by persons skilled in the art.
(Proanthocyanidins is the general name of an extensive big class polyphenolic substance that exists in the vegetable kingdom PACs) to procyanidin, extensively is present in the natural plants, in Fructus Vitis viniferae, Fructus Hippophae, Cortex Pini and huckleberry plant.Originally system belongs to condensed tannin or flavanol compound.Along with the raising of isolation identification technology with to the further investigation and the heightened awareness of this type of material, now become a big class material of taking the course of its own and be referred to as procyanidin.PACs is the polymer of flavonol, also claims condensed tannin.Promptly by the flavan-3-alcohol (flavan-3-ols) or the flavane-3 of varying number, the 4-glycol is polymerized.The simplest procyanidin is the dimer of catechin or epicatechin or catechin and epicatechin formation, also has trimer, the tetramer etc. in addition more than ten aggressiveness.Confirmed that procyanidin has the various biological function, comprising the function of antibacterium adhesion.Procyanidin in some plants for example is derived from the A type bonding (A-type linkage) that procyanidin had of cranberry, and has the effect that stronger anti-urinary tract P-pili escherichia coli attach to urothelial cell owing to its particular structure.
Pachyman is known to be a kind of composition of Poria.Pachyman can obviously increase phagocytosis of macrophages, strengthens the T lymphocyte function, but and the activating complement C3 and the B factor, intensive antitumor action is arranged.
Herba Taraxaci has another name called Herba crotalariae albidae, is the Compositae herbaceos perennial, has very high nutritive value.
Those skilled in the art can adopt prior art to obtain procyanidin from various known natural plants source as extractions such as peanut skin, Cortex Pini, Semen Vitis viniferae, or obtain to be suitable for the procyanidin of the present composition easily by various commercially available approach.Equally, those skilled in the art can adopt prior art to extract the pachyman that obtains pachyman or obtain to be suitable for the present composition by various commercially available approach easily from the sporophore of Poria or skin.Pachyman can be different withdrawal ratio.Therefore, the present invention is for the concrete acquisition mode and the source and without particular limitation of procyanidin, Herba Taraxaci and pachyman.
Yet in a good especially embodiment, procyanidin of the present invention can obtain from cranberry.Cranberry is the natural fruit a kind of with health role that originates in the North America, cultivates in the marshland of continent, Massachusetts, New Jersey, Wei Sikangxin, Washington and Oregon or the impregnated meadow of rainwater in 1816.In recent years the cranberry of discovering can suppress sticking of helicobacter pylori and gastric epithelial cell.It is that the procyanidin that exists in the cranberry has been realized this inhibition effect that the present inventor infers.Therefore, a particularly preferred method is among the present invention, can adopt fruit juice or the cranberry extract of cranberry.Cranberry fruit juice or cranberry extract can be buied by commercially available approach, or obtain with conventional means (for example can be according to Phytochemistry 2005; The method of describing among the 66:2281-2291 is extracted and is obtained).In another embodiment, procyanidin of the present invention can extract acquisition by this area conventional method from Semen Vitis viniferae.
Term used herein " contains ", " comprising ", " comprising " can exchange use when being used to characterize compositions of the present invention, its implication is represented in the said composition except listed component such as procyanidin, Herba Taraxaci and pachyman, can also contain some other component of not pointing out, as long as the existence of these components does not have adverse influence for the required effect that reaches of the present composition.At one more preferably in the embodiment, compositions of the present invention is made up of procyanidin, Herba Taraxaci and pachyman.Herein, term " by ... form " represent that then compositions of the present invention is made up of pointed concrete component, do not have other components, but it can have the impurity of content in common scope.
Procyanidin, Herba Taraxaci and pachyman content separately can be determined according to the experiment of routine by those skilled in the art in the application's compositions.Contain in the compositions of the present invention more preferably in the embodiment at one, calculate the pachyman of the procyanidin of 0.01-99%, the Herba Taraxaci of 0.01-99% and 0.01-99% (and three's summation is no more than 100%) with the weight of compositions., contain in the described compositions more preferably in the embodiment at another, calculate the pachyman of the procyanidin of 1-90%, the Herba Taraxaci of 1-90% and 1-90% (and three's summation is no more than 100%) with the weight of compositions.In addition, each components contents also can concern by the weight fraction between each component and represents in the present composition.In one embodiment, contain 0.02-100 weight portion procyanidin, 0.1-100 weight portion pachyman and 1-100 weight portion Herba Taraxaci in the compositions of the present invention.In embodiment more preferably, contain 2-80 weight portion procyanidin, 0.5-80 weight portion pachyman and 2-80 weight portion Herba Taraxaci in the compositions of the present invention.In a more preferred embodiment, contain 5-50 weight portion procyanidin, 1-50 weight portion pachyman and 5-50 weight portion Herba Taraxaci in the compositions of the present invention.When also containing other component except that procyanidin, pachyman and Herba Taraxaci in the compositions of the present invention when (being used to prevent and/or treat the active ingredient of helicobacter pylori infections) as pharmaceutically acceptable carrier or other, the total amount of above-mentioned procyanidin, pachyman and Herba Taraxaci (weight) is the 0.01-99.9% (weight) of present composition weight, more preferably for 0.1-90% (weight), be more preferably 0.5-80% (weight), 1-50% (weight) preferably also.
When adopting cranberry fruit juice, compositions of the present invention is preferably liquid form.The concentration of cranberry fruit juice can (more preferably being 5-80%) change between 1-100%, also can be its concentrated juice (in concentrating 50 times).Compositions of the present invention can 0.1-100 restrains pachyman and 1-100 gram Herba Taraxaci makes easily by adding in every liter of cranberry fruit juice (concentration is 10-50%).In a preferred embodiment, should contain 0.5-80 gram pachyman and 2-80 gram Herba Taraxaci in every liter of cranberry fruit juice (concentration is 10-50%).Also want in the embodiment preferred at one, should contain 1-50 gram pachyman and 5-50 gram Herba Taraxaci in every liter of cranberry fruit juice (concentration is 10-50%).Said composition can be taked the form of concentrate, or can further dilute taking previous crops.
It will be appreciated by those skilled in the art that essence of the present invention is to find to have produced between procyanidin (cranberry extract or cranberry fruit juice), pachyman and three components of Herba Taraxaci the effect of the opposing helicobacter pylori of remarkable excellence.Therefore, only need comprise procyanidin (cranberry extract or cranberry fruit juice), pachyman and the Herba Taraxaci that treats and/or prevents the helicobacter pylori infections effective dose in the compositions of the present invention.Those skilled in the art can determine suitable consumption according to the multi-form and different purposes of compositions fully, and therefore scope of the present invention should not be subjected to the restriction of the concrete content of each component.
At one more preferably in the embodiment, when compositions of the present invention during as medicine, said composition also can contain pharmaceutically acceptable carrier." pharmaceutically acceptable carrier " used herein should with each component compatibility in the present composition, can not reduce pharmaceutical composition with its blend under normal conditions significantly in the effect that treats and/or prevents aspect the helicobacter pylori.The object lesson that can be used as some materials of pharmaceutically acceptable carrier or its component is a saccharide, as lactose, dextrose plus saccharose; Starch is as corn starch and potato starch; Cellulose and derivant thereof are as sodium carboxymethyl cellulose, ethyl cellulose and methylcellulose; The tragakanta powder; Fructus Hordei Germinatus; Gelatin; Talcum; Kollag is as stearic acid and magnesium stearate; Calcium sulfate; Vegetable oil is as Oleum Arachidis hypogaeae semen, Oleum Gossypii semen, Oleum sesami, olive oil, Semen Maydis oil and cupu oil; Polyhydric alcohol is as propylene glycol, glycerol, Sorbitol, mannitol and Polyethylene Glycol; Alginic acid; Emulsifying agent is as Tween; Wetting agent is as sodium lauryl sulfate; Coloring agent; Flavoring agent; Tablet agent, stabilizing agent; Antioxidant; Antiseptic; Apirogen water; Deng oozing saline solution; With phosphate buffer etc., wherein more preferably carrier is normal saline or phosphate buffered saline (PBS).
All right and the other medicines coupling of the combination of procyanidin of the present invention (especially cranberry extract or cranberry fruit juice), pachyman and Herba Taraxaci.Therefore, in another embodiment preferred, also can contain medicine, antibiotic and/or acid secretion inhibitors that other has control helicobacter pylori effect in the compositions of the present invention.
When compositions of the present invention is used as medicine, compositions of the present invention can be made various medically acceptable dosage forms, and can by the doctor according to patient's kind, age, body weight and roughly factor such as disease condition, administering mode determine the useful dosage of patient is used.Preparation of the present invention is meant multiple liquid dosage forms such as oral liquid, injection, sublingual administration agent, or is prepared into multiple other dosage forms such as tablet, capsule by appropriate excipients in addition.In embodiment more preferably, compositions of the present invention can be made peroral dosage form, for example includes, but are not limited to oral liquid, tablet, capsule, electuary, granule, pill, powder etc.
Each active component in the present composition as procyanidin, pachyman and Herba Taraxaci, can exist in same preparation or the dosage form, also can separately be present in the different dosage form of same medicine, before using by carrying out administration after the user mixing.
More preferably in the embodiment, the combination of the procyanidin among the present invention, pachyman and Herba Taraxaci can add in the food, therefore also can contain other food component in the compositions of the present invention at another.In this case, the any solid that compositions of the present invention can be accepted on can food prepared therefrom or the form of liquid, for example include, but are not limited to: 1) milk product: cheese, butter, milk and other milk beverage, be coated in soft food and milk product incorporating, ice cream and yoghourt on the bread; 2) based on the goods of fat: as Margarine, be coated in soft food, mayonnaise, shortening, culinary art and frying oils and flavoring agent on the bread; 3) based on the goods of corn: as bread, wheaten food, cake, cookies etc.; 4) confection: as chocolate, confection, chewing gum, dessert, sherbet, sugar-coat and other stuffing material; 5) beverage: alcohol type or non-alcoholic drink, comprising can happy other soft drink, fruit juice, dietary supplement; 6) other goods: as the food of egg and egg products, processing such as soup, ready-formed wheaten food sauce, ready-made meals, teabag, animal feed etc.Compositions of the present invention can be with such as mixing, inculcate, injection, blending, dispersion, emulsifying, immersion, spraying and method such as knead is not done further to change and directly mix in the food.Perhaps, said composition can directly be applied to before ingesting on the food or be added in the beverage by consumer.These are simple and economic mode of movements.
More preferably in the embodiment, the combination of the procyanidin among the present invention, pachyman and Herba Taraxaci can add in the health product, therefore also can contain other consumer care ingredients in the compositions of the present invention at another.
Compositions of the present invention can prepare by each active component is mixed easily.For example, each component can be added in the pharmaceutically acceptable carrier (as water) successively or simultaneously, stir then, make the compositions of homogeneous.In addition, under the situation that adopts cranberry fruit juice, can directly pachyman and Herba Taraxaci directly be added in the cranberry fruit juice.As mentioned above, procyanidin (or cranberry fruit juice), pachyman and the Herba Taraxaci that is applicable to the present composition all can make by commercially available approach.For the ease of absorbing or other purpose, also can carry out some pretreatment to various active component raw materials.For example, water dilutes the cranberry concentrated juice, to be deployed into suitable concentration.Herba Taraxaci can be in advance through steps such as conventional oven dry, pulverizing.
The present invention relates to the application of described compositions on the other hand, that is, described compositions can be used to prepare medicine, food or health product.
As confirming in the embodiment of the present application that the helicobacter pylori infections effect can be treated and prevent to compositions of the present invention significantly.Therefore, estimate that compositions of the present invention can be used for preventing and/or treating the disease relevant with helicobacter pylori infections, for example include, but are not limited to: gastritis (as chronic gastritis, superficial gastritis, activeness and persistence chronic atrophic gastritis), peptic ulcer (as duodenal ulcer and gastric ulcer), gastric cancer etc.
Compositions of the present invention can be determined by means such as routine test such as rapid urease test, slice culture and histological examinations the effect of anti-helicobacter pylori.RUT test kit in the market all be with phenol red as indicator, come the qualitative detection helicobacter pylori by reagent by xanthochromia is red, be to be used for clinically mostly, and can not reflect the amount of helicobacter pylori in the gastric mucosa.When being used for research work, when especially carrying out work such as drug screening, these test kits obviously all can not be satisfied the demand.The present invention uses a kind of phenol indigo plant of using as indicator, reagent is by xanthochromia indigo plant when positive, not only change color is more remarkable, be beneficial to perusal, and can come quantitative response to go out the helicobacter pylori infections amount by the absorbance of measuring under the specific wavelength, be not only applicable to research work, and can be used for clinical.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Unless description is arranged in addition, enforcement of the present invention will be adopted the conventional means known to those skilled in the art or carry out according to the description that the reagent manufacturer is provided.
Formulation example 1:
Procyanidin: (Phytochemistry 2005 according to document; Procyanidin is extracted in description 66:2281-2291) from cranberry.
Pachyman (withdrawal ratio is 10: 1, and promptly the per kilogram pachyman is extracted by 10 kilograms of Poria and obtains): available from Zhejiang grid Pharmaceutical edible fungi polysaccharide company limited.
Herba Taraxaci:, cross 100 mesh sieves with Herba Taraxaci oven dry, pulverizing.
Add 0.2 gram procyanidin in every liter of distilled water or/and 1.2 gram pachymans or/and 18 gram Herba Taraxacis, fully mixing makes the reagent of the present invention and the contrast agents that are used for following test example 1 and test example 2 respectively.
Formulation example 2:
Procyanidin: (Phytochemistry 2005 according to document; Procyanidin is extracted in description 66:2281-2291) from cranberry.
Pachyman: with used identical of formulation example 1, available from Zhejiang grid Pharmaceutical edible fungi polysaccharide company limited.
Herba Taraxaci:, cross 100 mesh sieves with Herba Taraxaci oven dry, pulverizing.
Add 0.2 gram procyanidin in every liter of distilled water or/and 6 gram pachymans or/and 18 gram Herba Taraxacis, fully mixing makes the reagent of the present invention and the contrast agents that are used for following test example 1 and test example 2 respectively.
Test example 1: mice helicobacter pylori infections treatment experiment
One. material and method:
1. bacterial strain and cultivation:
The helicobacter pylorus bacteria strain that this institute is used be Sydney strain (Sydney Strain, SS1).Through the liquid culture amplification, centrifugal 10min under the 5000rpm collects antibacterial.Thalline is suspended from the improved broth medium, utilizes the spectrophotometric determination bacterial concentration, work as OD
660=1 o'clock, bacterial concentration was 10
8/ ml.Concentration is adjusted to 2 * 10
8/ ml is used to inoculate mice.
2. animal:
Laboratory animal is no special pathogen (S.P.F) level C57BL/6 mice, and in 6～8 ages in week, 18～20g entrusts Chinese Academy of Sciences's Shanghai Experimental Animal Center to raise.
3. treatment experiment
Per os is irritated stomach 1 * 10
8The CFU helicobacter pylori, the next day 1 time, totally 3 times.2 weeks from each group, put to death 1 mice respectively at random behind last 1 filling stomach, observe the helicobacter pylori infections situation.Mice is divided into 12 groups then, per os is irritated stomach and is given procyanidin, pachyman, Herba Taraxaci, pachyman respectively and add Herba Taraxaci, procyanidin and add that pachyman, procyanidin add Herba Taraxaci, procyanidin adds pachyman and Herba Taraxaci (is 0.5ml, once-a-day, totally 30 days).The 1st group for not treating matched group.24h after the various processing and put to death mice after 4 weeks respectively observes the helicobacter pylori infections situation.
4. the detection of helicobacter pylori infections:
The experimental mouse stomach along big curved vertically incision, vertically is cut into onesize 3 parts, and 1 part is directly carried out the sxemiquantitative urease test, put into physiological saline solution for 1 part, be used for antibacterial culturing, 1 part places 10% formalin fixing, is used for histopathology (Giemsa and H﹠amp; E dyeing) detect.
The sxemiquantitative urease test: wherein 1 part of gastric mucosa is put into 500 μ l urease detectable (50g carbamide, 0.3g phenol indigo plant adds 0.1% potassium dihydrogen phosphate 1000ml, pH value 5.0) in, room temperature was placed after 5 hours, detected its absorbance (OD) value at 630nm wavelength place.This value becomes positive correlation with the Hp infective dose, but so this result's quantitative response go out the infective dose of gastric mucosa Hp.Under the situation that Hp is not removed fully, also can demonstrate prevention effect.
Cultivate: 1 part of gastric mucosa mucosa is faced down, evenly coat on the blood agar plate, cultivated 5-7 days in 37 ℃ of little oxygen environment, determine whether to be helicobacter pylori through smear Gram, urease, catalase, oxidase test.
5. criterion
Meet entirely and be the helicobacter pylori feminine gender when following 3.(1) mean of infecting mouse OD630nm (mean)+2 times standard deviation (SD) of rapid urease test: OD630nm＜not; (2) cultivate feminine gender; (3) antibacterial is not seen in histological examination.
Therapeutic effect comes overall merit with clearance rate (i.e. the percentage ratio of treatment back 24h helicobacter pylori removing), eradication rate (i.e. the percentage ratio of the helicobacter pylori eradication in treatment 4 weeks of back) and sxemiquantitative urease test result (OD630nm).Preventive effect is come overall merit with prevention rate and sxemiquantitative urease test result (OD630nm).
Two. the result
1. treatment back 24h mice helicobacter pylori infections situation (clearance rate) sees the following form 1.
Table 1. treatment back 24h mouse infection situation
A compares with matched group, p≤0.01 (two-tailed Fisher ' s Exact Probability Test, the accurate probability inspection of two tail Fisher ' s)
B compares with the 2nd, 3,4,5 group, p≤0.01 (two-tailed Fisher ' s Exact Probability Test)
*Compare p＜0.05 (ANOVA, variance analysis) with matched group
*Compare with matched group, p＜0.01, the 11,12 group is compared with the 2nd, 3,4,5 group, and p＜0.05 (ANOVA) treatment back 4 all mouse infection situations (eradication rate) see the following form 2.
Table 2. treatment back 4 all mouse infection situations
A compares with matched group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
B compares with matched group, p＜0.01 (two-tailed Fisher ' s Exact Probability Test)
C compares with the 2nd group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
Compare p＜0.01 (two-tailed Fisher ' s Exact Probability Test) with the 3rd, 4,5 group
*Compare p＜0.05 (ANOVA) with matched group
*Compare with matched group, p＜0.05 (ANOVA) group compared in p＜0.01, the 11,12 with the 2nd, 3,4,5 group
According to The above results as can be seen, compare with single result with procyanidin, pachyman, Herba Taraxaci, the combination of procyanidin, pachyman and Herba Taraxaci has the effect for the treatment of helicobacter pylori infections more significantly.
Test example 2: mice helicobacter pylori infections prevention experiment
One. material and method
1-2. with test example 1
3. prevention experiment
Mice is divided into 12 groups, giving procyanidin, pachyman, Herba Taraxaci, pachyman in 1-60 days respectively adds Herba Taraxaci, procyanidin and adds that pachyman, procyanidin add Herba Taraxaci, procyanidin adds pachyman and Herba Taraxaci (is 0.5ml, once-a-day).The 1st group for not preventing matched group.Wherein giving helicobacter pylori on the the 42nd, 44,46 day attacks.Put to death mice on the 61st day and identify the helicobacter pylori infections situation.
4-5. with test example 1.
Two. the result
1. the prevention experimental result sees the following form 3.
Table 3. prevention mouse infection situation
A compares with matched group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
B compares with matched group, p＜0.01 (two-tailed Fisher ' s Exact Probability Test)
C compares with the 2nd group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
Compare p＜0.01 (two-tailed Fisher ' s Exact Probability Test) with the 3rd, 4,5 group
*Compare p＜0.05 (ANOVA) with matched group
*Compare with the 1st, 3,4,5 group, p＜0.05 (ANOVA) compared in p＜0.01 with the 2nd group
As can be seen from Table 3, compare with single result with procyanidin, pachyman, Herba Taraxaci, the combination of procyanidin, pachyman and Herba Taraxaci has the effect of preventing helicobacter pylori infections more significantly.
Formulation example 3
Cranberry fruit juice: concentration is 17% (v/v), adds the entry allotment by cranberry concentrated juice (purchasing the Ocean Spray company in the U.S.).
Pachyman: identical with formulation example 1, available from Zhejiang grid Pharmaceutical edible fungi polysaccharide company limited.
Herba Taraxaci:, cross 100 mesh sieves with Herba Taraxaci oven dry, pulverizing.
Do not add or add 1.2 gram pachymans or/and 18 gram Herba Taraxacis in every liter of cranberry fruit juice or distilled water, fully mixing makes the reagent of the present invention and the contrast agents that are used for following test example 3 and test example 4 respectively.
Formulation example 4
Cranberry fruit juice: concentration is 17% (v/v), is allocated by cranberry concentrated juice (purchasing the Ocean Spray company in the U.S.).
Pachyman: identical with formulation example 1, available from Zhejiang grid Pharmaceutical edible fungi polysaccharide company limited.
Herba Taraxaci:, cross 100 mesh sieves with Herba Taraxaci oven dry, pulverizing.
Do not add or add 6 gram pachymans or/and 18 gram Herba Taraxacis in every liter of cranberry fruit juice or distilled water, fully mixing makes the reagent of the present invention and the contrast agents that are used for following test example 3 and test example 4 respectively.
Test example 3: mice helicobacter pylori infections treatment experiment
One. material and method:
1. bacterial strain and cultivation:
The helicobacter pylorus bacteria strain that this institute is used be Sydney strain (Sydney Strain, SS1).Through liquid culture amplification, 5000rpm, the centrifugal collection of 10min antibacterial.Thalline is suspended from the improved broth medium, utilizes the spectrophotometric determination bacterial concentration, work as OD
660=1 o'clock, bacterial concentration was 10
8/ ml.Concentration is adjusted to 2 * 10
8/ ml is used to inoculate mice.
2. animal:
Laboratory animal is no special pathogen (S.P.F) level C57BL/6 mice, and in 6～8 ages in week, 18～20g entrusts Chinese Academy of Sciences's Shanghai Experimental Animal Center to raise.
3. treatment experiment
Per os is irritated stomach 1 * 10
8The CFU helicobacter pylori, the next day 1 time, totally 3 times.2 weeks from each group, put to death 1 mice respectively at random behind last 1 filling stomach, observe the helicobacter pylori infections situation.Mice is divided into 12 groups then, per os is irritated stomach and is given cranberry juice, pachyman, Herba Taraxaci, pachyman respectively and add Herba Taraxaci, cranberry juice and add that pachyman, cranberry juice add Herba Taraxaci, cranberry juice adds pachyman and Herba Taraxaci (is 0.5ml, once-a-day, totally 30 days).The 1st group for not treating matched group.24h after the various processing and put to death mice after 4 weeks respectively observes the helicobacter pylori infections situation.
4. the detection of helicobacter pylori infections:
The experimental mouse stomach along big curved vertically incision, vertically is cut into onesize 3 parts, and 1 part is directly carried out rapid urease test, put into physiological saline solution for 1 part, be used for antibacterial culturing, 1 part places 10% formalin fixing, is used for histopathology (Giemsa and H﹠amp; E dyeing) detect.
The sxemiquantitative urease test: wherein 1 part of gastric mucosa is put into 500 μ l urease detectable (50g carbamide, 0.3g phenol indigo plant adds 0.1% potassium dihydrogen phosphate 1000ml, pH value 5.0) in, room temperature was placed after 5 hours, detected its absorbance (OD) value at 630nm wavelength place.This value becomes positive correlation with the Hp infective dose, but so this result's quantitative response go out the infective dose of gastric mucosa Hp.Under the situation that Hp is not removed fully, also can demonstrate prevention effect.
Cultivate: 1 part of gastric mucosa mucosa is faced down, evenly coat on the blood agar plate, cultivated 5-7 days in 37 ℃ of little oxygen environment, determine whether to be helicobacter pylori through smear Gram, urease, catalase, oxidase test.
5. criterion
Meet negative entirely when following 3.(1) mean of infecting mouse OD630nm (mean)+2 times standard deviation (SD) of rapid urease test: OD630nm＜not; (2) cultivate feminine gender; (3) antibacterial is not seen in histological examination.
Therapeutic effect comes overall merit with clearance rate (i.e. the percentage ratio of treatment back 24h helicobacter pylori removing), eradication rate (i.e. the percentage ratio of the helicobacter pylori eradication in treatment 4 weeks of back) and sxemiquantitative urease test result (OD630nm).Preventive effect is come overall merit with prevention rate and sxemiquantitative urease test result (OD630nm).
Two. the result
Treatment back 24h mouse infection situation sees the following form 4.
Table 4. treatment back 24h mouse infection situation
A compares with matched group, p＜0.01 (two-tailed Fisher ' s Exact Probability Test)
B compares with the 2nd group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
Compare p＜0.01 (two-tailed Fisher ' s Exact Probability Test) with the 3rd, 4,5 group
*Compare p＜0.05 (ANOVA) with matched group
*Compare with the 1st, 3,4,5 group, p＜0.01 (ANOVA) compares with the 2nd group, and p＜0.05 (ANOVA) treatment back 4 all mouse infection situations see the following form 5, Fig. 1 and Fig. 2.
Table 5. treatment back 4 all mouse infection situations
A compares with matched group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
B compares with matched group, p＜0.01 (1wo-tailed Fisher ' s Exact Probability Test)
C compares for the 2nd group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
Compare p＜0.01 (two-tailed Fisher ' s Exact Probability Test) with the 3rd, 4,5 group
*Compare p＜0.05 (ANOVA) with matched group
*Compare with the 1st, 3,4,5 group, p＜.01 (ANOVA) compares p＜0.05 (ANOVA) with the 2nd group for the 11st, 12 group
In addition, as seen from Figure 1, matched group is infecting helicobacter pylori after 8 weeks, visible a large amount of helicobacter pylori field planting (Giemsa dyeing) in the gastric mucosa.And in Fig. 2,, do not see helicobacter pylori (Giemsa dyeing) with 4 weeks behind the combined therapy of cranberry fruit juice of the present invention, pachyman (6 gram) and Herba Taraxaci.
Can know that according to above-mentioned result of the test compare with single result with cranberry fruit juice, pachyman, Herba Taraxaci, the combination of cranberry fruit juice, pachyman and Herba Taraxaci has the effect of preventing helicobacter pylori infections more significantly.
Test example 4: mice helicobacter pylori infections prevention experiment
One. material and method
1-2. with test example 3
3. prevention experiment
Mice is divided into 12 groups, giving cranberry juice, pachyman, Herba Taraxaci, pachyman in 1-60 days respectively adds Herba Taraxaci, cranberry juice and adds that pachyman, cranberry juice add Herba Taraxaci, cranberry juice adds pachyman and Herba Taraxaci (is 0.5ml, once-a-day).The 1st group for not preventing matched group.Wherein giving helicobacter pylori on the the 42nd, 44,46 day attacks.Put to death mice on the 61st day and identify the helicobacter pylori infections situation.
4-5. with test example 3
Two. the result
The prevention experimental result sees the following form 6.
Table 6. prevention mouse infection situation
A compares with matched group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
B compares with matched group, p＜0.01 (two-tailed Fisher ' s Exact Probability Test)
C compares with the 2nd group, p＜0.05 (two-tailed Fisher ' s Exact Probability Test)
Compare p＜0.01 (two-tailed Fisher ' s Exact Probability Test) with the 3rd, 4,5 group
*Compare p＜0.05 (ANOVA) with matched group
*Compare with matched group, p＜0.01 (ANOVA) compares p＜0.05 (ANOVA) with the 2nd, 3,4,5 group for the 11st, 12 group
The result learns from table 6, compares with single result with cranberry fruit juice, pachyman, Herba Taraxaci, and the combination of cranberry fruit juice, pachyman and Herba Taraxaci has the effect of preventing helicobacter pylori infections more significantly.
In addition, except the foregoing description, the inventor is also to having prepared the compositions of procyanidin commercially available prod from Semen Vitis viniferae (available from last Haikang nine chemical companies) and pachyman, Herba Taraxaci according to the same procedure of above-mentioned formulation example and test example.Result of the test proves, should obtain too from the combination of the procyanidin of Semen Vitis viniferae and pachyman, Herba Taraxaci than single with the significantly excellent treatment of grape pip procyanidin, pachyman or Herba Taraxaci and the effect of prevention helicobacter pylori infections.
Although the invention describes concrete example, having a bit is significantly to those skilled in the art, promptly can do various variations and change to the present invention under the premise without departing from the spirit and scope of the present invention.Therefore, claims have covered all these changes within the scope of the present invention.It is for referencial use that this paper is all included in all publications, patent and the patent application that this paper quotes in.
Claims (10)
1. a compositions is characterized in that, described compositions comprises procyanidin, pachyman and the Herba Taraxaci that treats and/or prevents the helicobacter pylori infections effective dose.
2. compositions as claimed in claim 1 is characterized in that described compositions also comprises pharmaceutically acceptable carrier.
3. compositions as claimed in claim 1, it is characterized in that, the content of described procyanidin is the 0.02-100 weight portion, the content of described pachyman is the 0.1-100 weight portion, the content of described Herba Taraxaci is the 1-100 weight portion, and the gross weight of described procyanidin, pachyman and Herba Taraxaci is the 0.01-99.9% weight of composition weight.
4. compositions as claimed in claim 1 is characterized in that, described compositions comprises cranberry or its extract, pachyman and Herba Taraxaci.
5. compositions as claimed in claim 4, it is characterized in that, described compositions comprises cranberry fruit juice, pachyman and Herba Taraxaci, and the ratio of each component is in the described compositions, contains 0.1-100 gram pachyman and 1-100 gram Herba Taraxaci in every liter of cranberry fruit juice.
6. compositions as claimed in claim 1 is characterized in that, described compositions has any dosage form of acceptable on bromatology or the pharmaceutics.
7. as each described compositions of claim 1-6, it is characterized in that the dosage form of described compositions is oral liquid, tablet, capsule, electuary, granule, pill, powder, teabag, beverage class or injection.
The described compositions of claim 1 preparation be used for the treatment of and/or the medicine of the disease that prevention is relevant with helicobacter pylori infections in application.
9. application as claimed in claim 8 is characterized in that, the wherein said disease relevant with helicobacter pylori infections is gastritis, peptic ulcer or gastric cancer.
10. the application of the described compositions of claim 1 in preparation food or health product.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103392808A (en) * | 2013-07-17 | 2013-11-20 | 江苏桂龙生物技术有限公司 | Fruit milk capable of adjunctively treating gastric ulcer |
| CN103768149A (en) * | 2014-01-15 | 2014-05-07 | 李海涛 | Lotus seedpod procyanidins soft capsule health care product for controlling gastric ulcer |
| RU2523009C2 (en) * | 2009-08-12 | 2014-07-20 | Иновативо Биомедицинас Технологийю Институтс, Сиа | Fruit-and-berry food product with anti-microbial effect against helicobacter pylori |
| CN104000194A (en) * | 2014-06-05 | 2014-08-27 | 南京瑞盈环保科技有限公司 | Lotus seedpod procyanidins soft capsules capable of resisting oxidation and enhancing immunity and manufacturing method |
| CN104724799A (en) * | 2015-03-02 | 2015-06-24 | 马彦丽 | Treatment agent for textile waste water chamber |
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| CN113080450A (en) * | 2021-04-13 | 2021-07-09 | 花开星辰化妆品(浙江)有限公司 | Health food for inhibiting and killing helicobacter pylori and preparation method thereof |
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| RU2523009C2 (en) * | 2009-08-12 | 2014-07-20 | Иновативо Биомедицинас Технологийю Институтс, Сиа | Fruit-and-berry food product with anti-microbial effect against helicobacter pylori |
| CN103392808A (en) * | 2013-07-17 | 2013-11-20 | 江苏桂龙生物技术有限公司 | Fruit milk capable of adjunctively treating gastric ulcer |
| CN103768149A (en) * | 2014-01-15 | 2014-05-07 | 李海涛 | Lotus seedpod procyanidins soft capsule health care product for controlling gastric ulcer |
| CN104000194A (en) * | 2014-06-05 | 2014-08-27 | 南京瑞盈环保科技有限公司 | Lotus seedpod procyanidins soft capsules capable of resisting oxidation and enhancing immunity and manufacturing method |
| CN104724799A (en) * | 2015-03-02 | 2015-06-24 | 马彦丽 | Treatment agent for textile waste water chamber |
| CN107501435A (en) * | 2017-09-30 | 2017-12-22 | 南京正宽医药科技有限公司 | The extracting method of pachymaran and its application |
| CN113080450A (en) * | 2021-04-13 | 2021-07-09 | 花开星辰化妆品(浙江)有限公司 | Health food for inhibiting and killing helicobacter pylori and preparation method thereof |
| CN114766665A (en) * | 2022-04-11 | 2022-07-22 | 湖北工业大学 | Application of pachyman procyanidin composition in inhibiting formation of AGEs (advanced glycation end products) in gastrointestinal tract |
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| CN101239091B (en) | 2011-07-06 |
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