CN117959385A

CN117959385A - Composition for regulating ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tract, and preparation method and application thereof

Info

Publication number: CN117959385A
Application number: CN202410144074.6A
Authority: CN
Inventors: 沈鹤霄; 李国龙; 熊云; 彭文聪; 张帆; 周先锋; 付广; 滕兆伟; 吕永玲
Original assignee: Maintain Biomedical Wuhan Co ltd
Current assignee: Maintain Biomedical Wuhan Co ltd
Priority date: 2024-02-01
Filing date: 2024-02-01
Publication date: 2024-05-03

Abstract

The invention discloses a composition for regulating the ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tracts, a preparation method and application thereof. The composition comprises 25% -45% of schisandra chinensis, 15% -40% of rhizoma polygonati, 15% -25% of ligusticum wallichii, 10% -20% of acerola cherry, 8% -20% of cordyceps sinensis, 5% -20% of sea buckthorn leaves, 1% -15% of radix astragali, 1% -10% of polygonatum odoratum, 0.5% -5% of Chinese yam and 0.1% -5% of apocynum venetum.

Description

Composition for regulating ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tract, and preparation method and application thereof

Technical Field

The invention relates to the technical field of medical health-care food, in particular to a composition for regulating the ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tracts, and a preparation method and application thereof.

Background

The large number of microorganisms exist in human intestinal tracts, and the group of microorganisms depend on the intestinal life of the human body, and simultaneously help the human body to complete various physiological and biochemical functions. Intestinal microorganisms not only play an important role in bridging between diet and host, but also regulate human health as well as metabolic products, so that the intestinal microorganisms are closely related to human health. The intestinal microorganisms reach microecological balance through dynamic physiological action with a host, and bacteria and endotoxin translocation in intestinal tracts are effectively prevented. When the normal microflora is influenced by the host and the external environment, the original balance is destroyed, the intestinal flora is dysregulated, and the variety, the quantity and the proportion of the normal intestinal flora are abnormally changed and are converted into a pathological combined state, so that the host is pathogenic.

Modern diets are often very fine and have a relatively high water content, and studies have shown that high sugar diets tend to increase the permeability of the intestinal tract, causing inflammatory reactions in the body, whereas proper supplementation with probiotics may improve inflammatory reactions, e.g. bacteriocins produced by lactobacillus acidophilus may inhibit the growth of bacteria causing intestinal inflammation, and may reduce the production of immunocompetent factors, reducing intestinal inflammation. The human body can reduce the capability of scavenging free radicals with the increase of age, and the excessive free radicals can cause the phenomena of protein denaturation, DNA damage and the like, and the studies show that bifidobacterium adolescentis, lactobacillus fermentum and the like can obviously increase the activity and the content of superoxide dismutase (SOD) in blood, thereby reducing the oxidation reaction of the free radicals and the damage to human cells, and having the functions of antioxidation, anti-aging and the like.

In view of this, the present invention has been made.

Disclosure of Invention

The invention aims to provide a composition for regulating the ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tracts, and a preparation method and application thereof, so as to solve the problems.

The invention is realized in the following way:

In a first aspect, the present invention provides a composition comprising schisandra chinensis, rhizoma polygonati, ligusticum chuanxiong, acerola cherry, cordyceps sinensis, seabuckthorn leaf, astragalus membranaceus, polygonatum odoratum, yam and apocynum venetum;

The composition comprises, by mass, 25% -45% of schisandra chinensis, 15% -40% of rhizoma polygonati, 15% -25% of ligusticum wallichii, 10% -20% of acerola cherry, 8% -20% of cordyceps sinensis, 5% -20% of sea buckthorn leaves, 1% -15% of astragalus mongholicus, 1% -10% of polygonatum odoratum, 0.5% -5% of Chinese yam and 0.1% -5% of apocynum venetum.

In a second aspect, the present invention provides the use of a composition as described above for ameliorating intestinal inflammation.

In some embodiments, the intestinal flora includes beneficial bacteria and conditionally pathogenic bacteria.

In some embodiments, the beneficial bacteria include at least one of bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum, and pediococcus pentosaceus.

In some embodiments, the beneficial bacteria are bifidobacterium adolescentis.

In some embodiments, the conditionally pathogenic bacteria include Egget-slow bacteria.

In a third aspect, the invention provides a preparation method of the composition, which comprises the step of uniformly mixing the components according to the proportion to obtain the composition.

In a fourth aspect, the invention provides the use of a composition as described above for the preparation of a food or pharmaceutical product for modulating the bifidobacterium adolescentis/conditionally pathogenic bacteria ratio in the intestinal tract.

In some embodiments, the enteropathogenic bacteria include Egget's disease causing bacteria.

In a fifth aspect, the present invention provides the use of a composition as described above for the preparation of a food or pharmaceutical product for promoting intestinal beneficial bacteria.

In some embodiments, the intestinal beneficial bacteria include at least one of bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum, and pediococcus pentosaceus.

In some embodiments, the beneficial bacteria are bifidobacterium adolescentis.

In a sixth aspect, the invention provides the use of a composition as defined above for the preparation of a food or pharmaceutical product for the inhibition of enteropathogenic bacteria.

The invention has the following beneficial effects:

According to the invention, the composition capable of regulating intestinal flora is prepared by combining shizandra berry, rhizoma polygonati, ligusticum chuanxiong, acerola cherry, cordyceps sinensis, sea buckthorn leaf, astragalus, polygonatum, chinese yam and apocynum venetum, and can improve the abundance of beneficial bacteria such as bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum and pediococcus pentosaceus, reduce the abundance of intestinal pathogenic bacteria such as Egget's bacteria, and maintain the steady balance of intestinal microorganisms by regulating the ratio of the beneficial bacteria to the intestinal pathogenic bacteria, so that related diseases such as intestinal inflammation are effectively improved.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

The invention provides a composition, which comprises, by mass, 25% -45% of shizandra berry, 15% -40% of rhizoma polygonati, 15% -25% of szechuan lovage rhizome, 10% -20% of acerola cherry, 8% -20% of cordyceps sinensis, 5% -20% of sea buckthorn leaf, 1% -15% of astragalus mongholicus, 1% -10% of polygonatum odoratum, 0.5% -5% of Chinese yam and 0.1% -5% of apocynum venetum.

Ligusticum wallichii contains various chemical components such as phthalides, terpenes, organic acids, esters thereof, alkaloids, polysaccharides and the like. Researches show that ligustrazine as the main alkaloid has the effects of resisting cerebral ischemia and protecting heart, and has the protection effect on hepatic ischemia reperfusion injury and hepatic fibrosis. Has antioxidant free radical, antiinflammatory, anticancer, platelet aggregation and thrombosis resisting, and nerve protecting effects. In addition, the ligusticum wallichii has obvious inhibition effect on a plurality of gram-negative intestinal bacteria, so the ligusticum wallichii has good inhibition effect on dysentery (Sonne), typhoid, vibrio cholerae, deformation, pyocyaneus, large intestine, paratyphoid bacillus and the like.

The acerola cherry contains rich vitamin C, wherein the vitamin C content in every 100 g of fruits reaches 1677 mg, has the effects of whitening, resisting oxidation and delaying aging, and also contains vitamin A, vitamin B1, vitamin B2, iron, calcium and other elements, and the vitamin C is an essential component for maintaining the life of a human body. The special anti-aging factor (SOD), namely superoxide dismutase, achieves the effects of resisting oxidation and aging by removing superoxide anion free radicals (O2-) harmful to organisms, and can remove superoxide anion free radicals to protect cells from injury, thereby achieving the effect of inhibiting cancer cells.

Cordyceps sinensis is used as a rare nourishing traditional Chinese medicine, and the main active ingredients of the Cordyceps sinensis comprise cordycepin, cordyceps sinensis polysaccharide, SOD enzyme and the like, and the Cordyceps sinensis has various effects of regulating immune system functions, resisting tumors, resisting fatigue, resisting oxidation, resisting aging, resisting inflammation, tonifying qi, tonifying kidney and the like. Cordycepin can be doped into RNA, has strong inhibition effect on nuclear Ploy (A) polymerase, thereby influencing mRNA formation and protein synthesis, and the mechanism is possibly related to cordycepin anti-tumor effect; cordycepin also exhibits potent antifungal, anti-HIV-type I virus and selective inhibition of Clostridium bacterial activity. Cordyceps polysaccharide can promote lymphocyte transformation, activate immunocompetent cells of organism, and has effects of resisting aging, resisting oxidation and virus, resisting blood coagulation, reducing blood sugar and blood lipid, etc. SOD enzyme as an active protein has antioxidant and antiaging effects.

The sea buckthorn leaves are rich in flavonoid substances, and the flavonoid substances have the effects of enhancing the tolerance of a human body, reducing the permeability of capillary walls, remarkably preventing and improving cardiovascular and cerebrovascular diseases and remarkably reducing cholesterol in blood. The sea buckthorn leaves contain a large amount of SOD superoxide dismutase, and can block free radicals generated by peroxidation of substances, so that the effects of weakening the activity of excessive free radicals, resisting oxidation, delaying aging and prolonging life are achieved. In addition, the sea buckthorn leaves are also rich in vitamin C, the vitamin C plays a vital role in maintaining the metabolism of the organism, the requirement of the human body on the vitamin C is larger than that of any one vitamin, and the vitamin C and the vitamin E can directly capture the effects of superoxide radicals and hydroxyl radicals, remove the free radicals in the body, regulate the activity of immune cells, improve the disease resistance of the human body and delay the aging of organs of the human body.

The astragalus contains multiple microelements such as saponin, flavone, polysaccharide, multiple amino acids, folic acid, selenium, zinc, copper, etc. Has effects in enhancing immunity, protecting liver, resisting aging, lowering blood pressure, and resisting bacteria. The component MAPS-5 in the astragalus polysaccharide has the effect of inducing T cell proliferation, and the astragalus polysaccharide can relieve the symptoms of type II diabetes insulin resistance, fatty liver and the like; the astragalus polysaccharide obviously improves the contents of superoxide dismutase (SOD), catalase (CAT) and glutathione in blood, and plays roles of antioxidation and anti-aging; the astragaloside can protect kidney oxidative damage induced by shock waves; the liver fibrosis progress is relieved by scavenging oxygen free radicals, inhibiting the production of TNF-alpha and TGF-beta in Kupffer liver cells; radix astragali has inhibiting effects on Bacillus dysenteriae, pneumococcus, hemolytic streptococcus A, B, C, golden yellow, lemon color, white staphylococcus, etc.

The rhizoma Polygonati Odorati contains steroid saponins, high isoflavone, polysaccharides and volatile oil, and has effects of lowering blood sugar, regulating immunity, resisting oxidation, resisting aging, and resisting tumor.

The rhizoma Dioscoreae is rich in starch, saponin, calcium, phosphorus, ferrum, carotene, vitamins, etc. Researches show that the Chinese yam can promote the brain to secrete dehydroepiandrosterone and has the effects of improving sleep and delaying aging. Rhizoma Dioscoreae is rich in cellulose, and can reduce cholesterol. The glycoprotein contained in rhizoma Dioscoreae is hydrolyzed into protein and saccharide in vivo, and has blood sugar lowering effect.

The effective components in herba Apocyni Veneti mainly comprise flavonoids, tannins, organic acids, alcohols, amino acids, etc., and have antiallergic, anticancer, radioprotective, antiaging effects.

By combining the above substances and optimizing the addition proportion thereof, a composition capable of regulating intestinal flora is obtained, which is specifically expressed as follows: improving the abundance of beneficial bacteria in the intestinal tract, reducing the abundance of conditional pathogenic bacteria, and further playing a role in maintaining the steady-state balance of intestinal microorganisms.

According to the experimental results of the inventors, beneficial bacteria that the composition of the present invention can modulate include bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum and pediococcus pentosaceus.

The bifidobacterium adolescentis (Bifidobacterium adolescentis) is a probiotic, and a sufficient amount of bifidobacterium adolescentis can effectively control the balance of flora in intestinal tracts of human bodies, can generate acetic acid and lactic acid, inhibit the harmful fermentation of pathogenic bacteria, promote gastrointestinal peristalsis and enable the human bodies to relieve constipation normally; can synthesize vitamin B group and amino acid, help human body digest and absorb, and improve Chemicalbook absorption of calcium ions; antagonism can be generated on external pathogenic bacteria, so that the external pathogenic bacteria cannot be planted in the intestinal tract; lactose can be effectively decomposed, and the absorption of human body to lactose is ensured; can protect human cells from being damaged by carcinogens; can protect liver and cardiovascular and cerebrovascular of human body; can improve immunity. Group Shujie Chen, jianmin Si and Tongyao Hou, published in Gut Microbes, report that bifidobacterium adolescentis alleviates DSS-induced chronic colitis by inducing a protective Th2/Treg response and remodelling the intestinal microbiota. The research result suggests that bifidobacterium adolescentis can improve the clinical treatment effect of inflammatory bowel disease. The effects of bifidobacterium adolescentis and lactobacillus acidophilus on maintaining the remission of ulcerative colitis are also verified by establishing a DSS-induced convalescence UC mouse model.

Lactobacillus salivarius (Lactobacillus salivarius) is a gram positive bacterium, and has no catalase and oxidase, and can produce lactic acid. The lactobacillus salivarius has a certain adsorption effect, and can well colonize and grow on the surface of the intestinal tract so as to play a role in immunity. Its immune effect is mainly manifested in both innate and acquired immune responses. Meanwhile, the antagonistic action and antiallergic effect of the compound on pathogenic bacteria are as follows: the bacteria can inhibit the growth of other bacteria by secreted antibiotics, bacteriocins and other substances in the growth and metabolism process of the lactobacillus colonitis in the intestinal tract, and compete with pathogenic bacteria for growth space so as to play an antagonistic role. Also, studies have shown that bacteriocins produced by lactobacillus salivarius in the gut under the same enzymatic reaction and secretion mechanism are highly diverse, providing a premise for their competitive advantage in the complex microbial system of the mammalian gut. In addition, lactobacillus salivarius has a certain positive effect on inflammatory bowel disease or irritable bowel syndrome.

Lactobacillus acidophilus (Lactobacillus acidophilus) is a probiotic widely existing in intestines and stomach of human and animals, has a probiotic effect on both human and animals, and can play a good health promoting effect when the number of live bacteria reaches a certain number. Lactic acid, hydrogen peroxide, bacteriocin, short chain fatty acid, enzyme and other substances produced by lactobacillus acidophilus can improve microecological balance of intestinal bacteria, stimulate specific or non-specific immunity, enhance body health, prevent certain diseases and delay aging. Lactobacillus acidophilus has also been reported to have therapeutic effects on acute ulcerative colitis.

Lactobacillus fermentum (Limosilactobacillus fermentum) is a heterologous fermentation strain, lactic acid and a large amount of acetic acid, ethanol and CO ₂ are produced after fermentation, and the lactobacillus fermentum has super strong intestinal cell adhesion capability and strong colonization, and effectively blocks adhesion of pathogenic bacteria (salmonella, escherichia coli, staphylococcus aureus, listeria and the like) in intestinal tracts. Can also generate a large amount of glutathione and natural antioxidant substances, and effectively prevent oxidative damage to intestinal tracts. Meanwhile, the prior art records that the lactobacillus fermentum has the functions of regulating intestinal functions and preventing colonitis.

Pediococcus pentosaceus (Pediococcuspentosaceus) is often distributed in wort and fermented plant materials such as pickled vegetables, kimchi, silage, etc. Currently, the Fang Dong team studied in depth the effect of the probiotic pediococcus pentosaceus CECT8330 on sodium dextran sulfate (DSS) -induced colitis in mice in animal models. This study demonstrates that Pediococcus pentosaceus CECT8330 protects DSS-induced colitis, regulates intestinal microbial composition and function, immune morphology and intestinal barrier function.

Through adjusting the abundance of the beneficial bacteria, the beneficial bacteria can act with the intestinal tract of a host together, so that intestinal inflammation can be reduced or improved synergistically.

According to the experimental results of the inventors, the conditional pathogenic bacteria that the composition of the present invention can inhibit include Egget's lentus bacteria.

In the invention, the preparation method of the composition comprises the step of uniformly mixing the components according to the proportion to obtain the composition.

Specifically, the preparation method of the composition comprises the following steps: the raw materials are crushed and then dissolved by adopting a buffer solution, and the buffer solution is uniformly mixed, wherein the types of the buffer solution in the invention are various and include but are not limited to one or more of PBS, phosphoric acid and citric acid.

According to the composition, the invention also provides a culture medium of the composition for regulating the ratio of the bifidobacterium adolescentis to the conditional pathogenic bacteria in the intestinal tract, which comprises a basal culture medium and the composition for regulating the ratio of the bifidobacterium adolescentis to the conditional pathogenic bacteria in the intestinal tract, and the preparation method of the culture medium is that the composition is added into the basal culture medium.

Wherein, the composition is dissolved by buffer solution and is prepared into a dissolving solution with the concentration of 4-6 percent, and then the dissolving solution is added into a basal medium, and the volume ratio of the dissolving solution to the basal medium is 1:1-1.5. The culture medium provided by the invention is used for culturing the fecal sample, so that the ratio of pathogenic bacteria in the fecal sample can be reduced.

The invention also provides various applications of the composition for regulating the ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tracts, which comprises the following steps:

Application of a composition for regulating the ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tracts in preparing food or medicine for promoting bifidobacterium adolescentis; application of composition for regulating ratio of bifidobacterium adolescentis to conditional pathogenic bacteria in intestinal tract in preparation of food or medicine for inhibiting conditional pathogenic bacteria or

Use of a composition for modulating the ratio of bifidobacterium adolescentis to conditionally pathogenic bacteria in the gut for the manufacture of a medicament or food for improving gut function, enhancing immunity; or is or

Use of a composition for modulating the ratio of bifidobacteria adolescentis to conditionally pathogenic bacteria in the intestinal tract in a method for directed fermentation of specific donor flora for fecal bacteria transplantation.

The features and capabilities of the present invention are described in further detail below in connection with the examples.

Fructus Schisandrae, rhizoma Polygonati, rhizoma Ligustici Chuanxiong, acerola cherry, cordyceps, folium Hippophae, radix astragali, rhizoma Polygonati Odorati, rhizoma Dioscoreae and herba Apocyni Veneti in the following examples are all purchased from SiAnoersted biotechnology Co.

Example 1

The present example provides a composition comprising the following components:

350g of schisandra chinensis, 200g of rhizoma polygonati, 140g of ligusticum wallichii, 100g of acerola cherry, 60g of cordyceps sinensis, 50g of sea buckthorn leaves, 40g of astragalus membranaceus, 20g of polygonatum odoratum, 20g of Chinese yam and 20g of apocynum venetum.

Example 2

The present example provides a composition comprising the following components:

280g of shizandra berry, 180g of rhizoma polygonati, 160g of szechuan lovage rhizome, 120g of acerola cherry, 70g of cordyceps sinensis, 50g of sea buckthorn leaf, 50g of astragalus root, 50g of fragrant solomonseal rhizome, 25g of Chinese yam and 15g of apocynum venetum.

Example 3

The present example provides a composition comprising the following components:

200g of shizandra berry, 200g of rhizoma polygonati, 200g of szechuan lovage rhizome, 100g of acerola cherry, 50g of cordyceps sinensis, 50g of sea buckthorn leaf, 50g of astragalus membranaceus, 50g of polygonatum odoratum, 40g of Chinese yam and 10g of apocynum venetum.

Example 4

The present example provides a composition comprising the following components:

450g of schisandra chinensis, 150g of rhizoma polygonati, 140g of ligusticum wallichii, 110g of acerola cherry, 60g of cordyceps sinensis, 40g of sea buckthorn leaves, 20g of astragalus membranaceus, 10g of polygonatum odoratum, 10g of Chinese yam and 10g of apocynum venetum.

Comparative example 1

The composition of this comparative example had the following ingredients: 305g of rhizoma polygonati, 215g of ligusticum wallichii, 155g of acerola cherry, 92g of cordyceps sinensis, 80g of sea buckthorn leaf, 62g of astragalus, 31g of polygonatum odoratum, 30g of Chinese yam and 30g of apocynum venetum.

Comparative example 2

The composition of this comparative example had the following ingredients: 435g of schisandra chinensis, 175g of ligusticum wallichii, 125g of acerola cherry, 75g of cordyceps sinensis, 63g of sea buckthorn leaves, 50g of astragalus mongholicus, 25g of polygonatum odoratum, 25g of Chinese yam and 25g of apocynum venetum.

Comparative example 3

The composition of this comparative example had the following ingredients: 405g of shizandra berry, 230g of rhizoma polygonati, 115g of acerola cherry, 75g of cordyceps sinensis, 60g of sea buckthorn leaves, 45g of astragalus, 25g of polygonatum odoratum, 23g of Chinese yam and 22g of apocynum venetum.

Comparative example 4

The composition of this comparative example had the following ingredients: 390g of schisandra chinensis, 220g of rhizoma polygonati, 155g of ligusticum wallichii, 68g of cordyceps sinensis, 55g of sea buckthorn leaves, 45g of astragalus mongholicus, 23g of polygonatum odoratum, 22g of Chinese yam and 22g of apocynum venetum.

Experimental example 1

This experimental example is an in vitro experiment performed on the compositions of examples 1-4 and comparative examples 1-4, and the specific procedure is as follows:

1. Fecal sample treatment

(1) PBS buffer solution preparation

Taking 1000ml as an example, the following medicines were weighed separately by electronic analysis and poured into a 1000ml beaker, comprising: KH ₂PO₄ 0.24g,Na₂HPO₄·12H₂ O2.90g,NaCl 8.00g,KCl 0.20g. The pH of the above solution was measured with a calibrated pH meter and ph=7.4±0.05 was adjusted with 0.1M HCl or NaOH. Then put into an autoclave at 121 ℃ for 15 minutes. After cooling to room temperature, the solution was stored in a refrigerator at 4℃for 6 months.

(2) Fecal sample treatment

Taking 10g as an example, the final concentration is 10%.

10G of fecal sample was weighed out with a one percent balance, an appropriate amount of the above PBS buffer was added to the centrifuge tube with an automatic pipette, and thoroughly mixed on a shaker. The completely mixed samples were then split equally into new 50ml centrifuge tubes, and an appropriate amount of PBS buffer was added to each tube.

Filtering in a biosafety cabinet after uniformly mixing, and sequentially passing diluted samples through filter screens of 20 meshes, 50 meshes, 100 meshes and 200 meshes. Collecting filtrate into a centrifuge tube, placing the centrifuge tube into a centrifuge, balancing, centrifuging at 6000G and 4 ℃ for 15min, and discarding the supernatant. After weighing the precipitate, the volume was fixed with PBS at a final concentration of 5% to obtain an intestinal microorganism sample.

2. Preparation of basic culture medium

Preparing a basal medium for culturing intestinal microorganisms: GAM medium, for example 1000ml. 60g of the modified GAM broth drug was weighed out with an electronic analytical balance and poured into a 1000ml beaker. 800ml of ultrapure water was measured with a measuring cylinder, poured into a beaker, put into a stirring rotor, put on a magnetic heating stirrer, stirred until completely dissolved, and fixed to 1000ml.121 ℃,15 minutes. After sterilization, the bottle cap is immediately screwed up and cooled to room temperature.

3. Preparation of the Mixed solution

The compositions of examples 1-4 and comparative examples 1-4 were dissolved in PBS buffer to a 5% mixture by mass.

4. Grouping and intervention

On a sterile bench, the above GAM medium was dispensed into glass tubes with a 1ml pipette, 2ml per tube. Next, the mixture of the compositions of examples 1 to 4 and comparative examples 1 to 4 was taken out by a 1ml pipette, and added to the above-mentioned glass tube containing GAM medium in an amount of 2ml per tube, to thereby obtain an experimental group. Meanwhile, a comparison group is also arranged, and the grouping is specifically as follows:

experimental group: 2mL of GAM culture medium+1 mL of mixed solution+1 mL of intestinal microbial sample 2mL of mixed solution+2 mL of PBS buffer solution;

Control group: 2mL of GAM medium+1 mL of intestinal microbial sample+3 mL of PBS buffer;

After each glass tube cover is confirmed to be screwed, the glass tube covers are transferred to an anaerobic box transfer box, and 84 disinfectant is needed for disinfection before the glass tube covers are placed. After the anaerobic tank is put into the anaerobic tank, the bottle cap is unscrewed to replace oxygen, and the replacement is needed for 12 hours.

Intestinal microbial samples obtained after the above treatment were inoculated into 1ml of each of a test group (composition-GAM medium) and a blank group (GAM medium-PBS buffer), respectively.

After culturing in an anaerobic box for 72 hours, the growth condition of the flora is observed, and the flora is photographed and retained.

The test and control samples were then centrifuged at 10000rpm for 3min, the supernatant discarded, and the abundance changes before and after intervention of part of the intestinal flora were analyzed for significance. The pellet was treated with liquid nitrogen and sent to wuhan ai kang biotechnology limited, and the abundance of intestinal flora in the test and control groups, respectively, was measured, with the abundance measurement results for a portion of the intestinal flora shown in table 1. And the abundance change before and after the intervention of part of intestinal flora is subjected to significance analysis. The results are analyzed as shown in Table 1:

TABLE 1 analysis of abundance changes before and after partial intestinal microbiota intervention

Note that: * P <0.05, #P <0.01.

The analysis results in table 1 show that: the composition can effectively change the abundance of intestinal microorganisms, wherein bifidobacterium adolescentis (Bifidobacterium adolescentis) is obviously increased, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum and pediococcus pentosaceus are also increased to a certain extent, and the significance of the slow egte bacteria (EGGERTHELLALENTA) is reduced, so that the composition-intervened experimental group is proved to be obviously enriched in the abundance of beneficial bacteria such as bifidobacterium adolescentis and the like, and the number of conditional pathogenic bacteria is reduced, so that the flora of the intestinal tract is regulated.

Experimental example 2

The experimental example verifies the inhibition effect of the composition on conditional pathogenic bacteria, namely Eggerthella lenta, and the specific conditions are as follows:

1. Fecal sample treatment

(1) PBS buffer solution preparation

(2) Fecal sample treatment

Taking 10g as an example, the final concentration is 10%.

2. Preparation of basic culture medium

3. Grouping and intervention

(1) Culture of Egget-slow bacteria

The Egget-slow bacteria (EGGERTHELLALENTA) are cultured anaerobically, and the strain is first inoculated into LB plate for activation and anaerobically cultured at 37 deg.C for 24 hr. Then transferring to an anaerobic liquid culture medium for culturing for 24 hours, wherein the culture medium is as follows: 15g/L of peptone, 5g/L of yeast powder, 5g/L of soybean powder, 5g/L of beef powder, 5g/L of glucose, 5g/L of sodium chloride, 3g/L of soluble starch, 0.5g/L of cysteine, 2.5g/L of potassium dihydrogen phosphate, 0.005g/L of hemin and 0.001g/L of vitamin K. pH 7.3 and 0.2.

(2) Identification of Egget's disease

Extracting genome DNA of the original bacteria after pure amplification culture of the original bacteria.

Forward primer: 27F (5'-AGAGTTTGATCATGGCTCAG-3')

Reverse primer: 1492R (5'-TACGGCTACCTTGTACGACTT-3')

A16 SrRNA gene PCR amplification experiment was performed. The reaction procedure was as follows, 96℃for 3min, 96℃for 30s, 58℃for 30s, 72℃for 1min, 35 cycles under this condition, 72℃for 10min, and after the PCR reaction was completed, 1% agarose was identified and the desired PCR product fragment was recovered using a gel recovery kit. After sequencing, the sequence was aligned with the 16S rRNA gene of known bacteria in the GeneBank database.

(3) Operating procedure

On a sterile bench, the above GAM medium was dispensed into glass tubes with a 1ml pipette, 2ml per tube. Then, a 1ml pipette was used to take a mixture of the composition of example 1, and the mixture was added to the above glass tube containing GAM medium, 2ml each, and as an experimental group, a condition control group and a blank group were further provided, and three groups were arranged in parallel, and the following groups were specifically defined:

experimental group: 3ml of GAM medium+1 ml of composition mixture+1 ml of Eggerthella lenta+1 ml of PBS buffer;

Condition control group: 3ml of GAM medium+1 ml of Eggerthella lenta+2 ml of PBS buffer;

blank group: 3ml GAM medium+3 ml PBS buffer

The intestinal microbial samples obtained after the treatment are respectively inoculated into an experimental group, a conditional control group and a blank control group, wherein each tube is 1ml.

The test and control samples were then centrifuged at 10000rpm for 3min, the supernatant was discarded, the pellet was treated with liquid nitrogen and sent to wuhan ai kang biosciences, ltd, to determine the abundance of intestinal flora in the test and control samples, respectively, i.e. the percentage of different intestinal microorganisms in all species detected, wherein the abundance determination of part of the intestinal flora is shown in table 2. And the change of abundance before and after the intervention of the slow Egget bacteria is subjected to significance analysis. The analysis results are shown in table 2:

TABLE 2 analysis of changes in abundance before and after Egget's disease prevention

Comparison blank group P < 0.01, comparison condition control group #p < 0.01.

The analysis results in Table 2 show that the composition of the invention has an inhibitory effect on Egget strain, as evidenced by a significant decrease in abundance after intervention by the composition.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition is characterized by comprising the components of shizandra berry, rhizoma polygonati, ligusticum chuanxiong, acerola cherry, cordyceps sinensis, sea buckthorn leaf, astragalus, polygonatum odoratum, chinese yam and apocynum venetum;

the composition comprises, by mass, 25% -45% of shizandra berry, 15% -40% of rhizoma polygonati, 15% -25% of ligusticum wallichii, 10% -20% of acerola cherry, 8% -20% of cordyceps sinensis, 5% -20% of sea buckthorn leaves, 1% -15% of astragalus mongholicus, 1% -10% of polygonatum odoratum, 0.5% -5% of Chinese yam and 0.1% -5% of apocynum venetum.

2. Use of a composition according to claim 1 for improving intestinal inflammation.

3. Use of a composition according to claim 1 for modulating the intestinal flora, wherein the intestinal flora comprises beneficial bacteria and conditionally pathogenic bacteria;

the beneficial bacteria comprise at least one of bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum and pediococcus pentosaceus;

Preferably, the beneficial bacteria are bifidobacterium adolescentis;

preferably, the conditionally pathogenic bacteria comprise Egget-slow bacteria.

4. The method for preparing the composition according to claim 1, wherein the composition is obtained by uniformly mixing the components according to a ratio.

5. Use of a composition according to claim 1 for the preparation of a food or pharmaceutical product for modulating the bifidobacterium adolescentis/conditionally pathogenic bacteria ratio in the intestinal tract.

6. The use according to claim 5, wherein the enteropathogenic bacteria comprise angete lentus.

7. Use of a composition according to claim 1 for the preparation of a food or pharmaceutical product for promoting intestinal beneficial bacteria.

8. The use according to claim 7, wherein the intestinal beneficial bacteria comprise at least one of bifidobacterium adolescentis, lactobacillus salivarius, lactobacillus acidophilus, lactobacillus fermentum and pediococcus pentosaceus;

preferably, the beneficial bacteria are bifidobacterium adolescentis.

9. Use of a composition according to claim 1 for the preparation of a food or pharmaceutical product for inhibiting enteropathogenic bacteria.

10. The use according to claim 9, wherein the enteropathogenic bacteria comprise angete lentus.