Composition for preventing and treating polycystic ovary syndrome and application thereof

Abstract

The present invention belongs to the field of biomedicine technology, and in particular, relates to a composition and application for preventing and treating polycystic ovary syndrome, comprising the following active components: Tripterygium wilfordii polysaccharide, Dendrobium tassel polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia muciniphila suspension. The present invention uses a female SD rat model induced by letrozole and a high-sugar and high-fat diet to prove the application of Akkermansia muciniphila in preventing polycystic ovary syndrome. The test method is safe and reliable, the test results are accurate, and the efficacy is significant. The present invention provides a composition capable of inhibiting, improving or treating polycystic ovary syndrome, which has a better effect than oral administration of Akkermansia muciniphila suspension alone, and the components of the composition of the present invention are safer, which solves the technical problem that the existing chemical synthetic drugs have side effects in the treatment of polycystic ovary syndrome, and has good application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a composition for preventing and treating polycystic ovary syndrome and its application. Background Art

[0002] Polycystic ovary syndrome (PCOS) is one of the most common endocrine and metabolic disorders, affecting 6% to 20% of premenopausal women. The main characteristics of PCOS are androgen excess, ovulatory dysfunction, and polycystic ovaries. In addition, PCOS is often accompanied by metabolic disorders, such as insulin resistance (IR), glucose intolerance, and obesity. The pathogenesis of PCOS is still unclear, and it may be closely related to factors such as inflammation, neuroendocrine, genetics, and environment. Although the etiology of PCOS remains unclear, a large amount of clinical evidence strongly supports the association between PCOS and various metabolic diseases, including obesity, abdominal obesity, insulin resistance, and obesity-induced cardiovascular risk factors. Among them, about 50% to 60% of PCOS patients are obese. Therefore, the treatment of PCOS has become a topic of common concern in the fields of gynecology and endocrinology. Although the study of PCOS has been carried out for a long time, its etiology and pathophysiological mechanism are still unclear.

[0003] Akkermansia muciniphila (Akk bacteria for short) is an important commensal bacterial genus in the human intestine. This Gram-negative anaerobic bacterium accounts for about 3% of the healthy human intestinal flora and relies on mucin in the intestinal mucus layer for survival. Akkermansia muciniphila plays a key role in maintaining healthy immune processes. Previous extensive studies have shown that the abundance of Akkermansia muciniphila is negatively correlated with inflammatory bowel disease, obesity, and type 2 diabetes; and induces intestinal adaptive immune responses during T cell subset homeostasis.

[0004] At present, the drugs for treating PCOS are mainly chemical synthetic drugs, such as insulin sensitizers, ovulation-inducing drugs, and anti-androgen drugs. Chemical synthetic drugs have good efficacy in treating PCOS, but they have side effects, such as gastrointestinal irritation and difficulty in implantation of fertilized eggs. Therefore, developing an effective and safe drug to replace chemical synthetic drugs is an important development direction for the treatment of PCOS in the future. Summary of the invention

[0005] The purpose of the present invention is to provide a composition for preventing and treating polycystic ovary syndrome and its application, so as to solve the problems raised in the above background technology.

[0006] In order to achieve the above technical objectives, the present invention provides a composition for preventing and treating polycystic ovary syndrome, comprising the following active components: Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension, wherein the weight ratio of Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension is 1-1.1:0.8:0.8:0.8:1-1.1.

[0007] As a further improvement, the Akkermansia muciniphila bacterial suspension contains Akkermansia muciniphila, and the strain number of Akkermansia muciniphila in the China Industrial Culture Collection Center CICC is CICC24917.

[0008] As a further improvement, the concentration of Akkermansia muciniphila in the Akkermansia muciniphila suspension is 1×10 9 CFU / mL.

[0009] As a further improvement, the culture method of the Akkermansia muciniphila suspension is as follows: 0.4 g of mucin and 1 mg of L-cysteine ​​are added to 200 mL of brain infusion medium BHI, cultured under anaerobic conditions, and then activated Akkermansia muciniphila is added, cultured at 37° C. for 24 to 48 hours, centrifuged to collect the cells, and resuspended in distilled water to prepare a suspension with a concentration of 1×10 9 The Akkermansia muciniphila suspension with a concentration of CFU / mL is stored at 4°C, wherein the concentration of mucin is 2 g / L and the concentration of L-cysteine ​​is 5 mg / L.

[0010] As a further improvement, the preparation method of the composition comprises the following steps:

[0011] S1. Extracting Tripterygium wilfordii polysaccharide and Dendrobium tassellensis polysaccharide from Tripterygium wilfordii and Dendrobium tassellensis respectively by water extraction and alcohol precipitation method, and separating and purifying Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide from Hericium erinaceus mycelium crude extract and Lycium barbarum extract respectively by AKTA protein purification method;

[0012] S2. Add the Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide to the Akkermansia muciniphila suspension in sequence and mix well.

[0013] The present invention also provides an application of a composition for preventing and treating polycystic ovary syndrome in preparing a drug for preventing and treating polycystic ovary syndrome.

[0014] Due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0015] (1) The present invention provides a composition for preventing and treating polycystic ovary syndrome. Experiments have found that Akkermansia muciniphila has a significant effect of inhibiting, improving or treating polycystic ovary syndrome, and can be used to prepare drugs for inhibiting, improving or treating polycystic ovary syndrome, solving the technical problem that existing chemical synthetic drugs have side effects in treating polycystic ovary syndrome, and has good application prospects.

[0016] (2) The present invention uses a female SD rat model induced by letrozole and a high-sugar and high-fat diet, and intervenes with Akkermansia muciniphila in the AKK group female SD rat model, thereby verifying that Akkermansia muciniphila can reduce the body weight of PCOS rats, effectively improve the insulin resistance of PCOS rats, reduce the increase in ovarian mass in PCOS rats and promote the recovery of ovarian mass in PCOS rats to normal levels, and effectively improve the ovarian lesions in PCOS rats. Akkermansia muciniphila can also protect the liver from damage related to hepatocellular fatty degeneration and cholesterolemia, and effectively increase the content of follicle-stimulating hormone in PCOS rats to normal levels.

[0017] (3) The present invention effectively verifies the application of Akkermansia muciniphila in treating polycystic ovary syndrome. The test method is safe and reliable, the test results are accurate, and the therapeutic effect is significant.

[0018] (4) By adding Tripterygium wilfordii polysaccharide to the composition, the growth of Akkermansia muciniphila is promoted, thereby solving the problem that oral Akkermansia muciniphila will be excreted from the body with feces in a short period of time, achieving the effect of long-term treatment and better treatment effect. In addition, the auxiliary addition of Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide can promote the balance of intestinal flora and the growth of probiotics, improve the intestinal environment, facilitate the function of Akkermansia muciniphila, and be more conducive to the treatment of polycystic ovary syndrome. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 is the content of serum sex hormone testosterone T in rats of the Control group and the Model group in Example 2;

[0021] Figure 2 is a diagram of the cell structure of the rat vaginal smear at different estrus cycles in Example 2;

[0022] Figure 3 It is a representative estrous cycle change diagram of the Control group and the Model group in Example 2;

[0023] Figure 4 is a graph showing changes in body weight of rats in each group after intragastric administration of Akkermansia muciniphila in Example 2;

[0024] Figure 5 is a graph showing changes in fasting insulin (INS) content of rats in each group in Example 2;

[0025] Figure 6 is a graph of the ovarian weights of rats in each group in Example 2;

[0026] Figure 7 2 is a pathological section diagram of the ovaries of the rats in each group in Example 2;

[0027] Figure 8 2 is a liver pathological section diagram of each group of rats in Example 2;

[0028] Fig. 9 is a graph of the follicle stimulating hormone (FSH) levels of rats in each group in Example 2;

[0029] Fig.10 is a graph showing the change in absorbance over time in Example 3;

[0030] Fig.11 is a graph showing changes in body weight of rats in each group in Example 6;

[0031] Fig.12 is a graph showing changes in fasting insulin (INS) content of rats in each group in Example 6;

[0032] Fig.13 This is a graph of the follicle stimulating hormone (FSH) levels of rats in each group in Example 6. DETAILED DESCRIPTION

[0033] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0034] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0035] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0036] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0037] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0038] The strain number of Akkermansia muciniphila in the present invention is CICC24917 in the China Industrial Culture Collection Center CICC. Unless otherwise specified, the Akkermansia muciniphila used in the following embodiments is Akkermansia muciniphila CICC24917. Example 1

[0039] Cultivation of Akkermansia muciniphila suspension: add 0.4 g mucin (2 g / L) and 1 mg L-cysteine ​​(5 mg / L) to 200 mL brain infusion medium BHI, culture under anaerobic conditions (10% hydrogen, 10% carbon dioxide, 80% nitrogen), then add activated Akkermansia muciniphila, culture at 37°C for 24-48 h, collect the cells by centrifugation, resuspend the cells in distilled water to make a concentration of 1×10 9 CFU / mL of Akkermansia muciniphila suspension was stored at 4°C. Example 2

[0040] The experiment of Akkermansia muciniphila improving polycystic ovary syndrome in female rats, the specific experimental method is as follows:

[0041] Modeling and intervention

[0042] (1) Animal grouping and modeling

[0043] Eighteen 6-week-old SPF female SD rats were selected, with a weight of 160-180 g. The 18 rats were randomly divided into three groups, with 6 rats in each group, namely the normal control group (Control group), the model control group (Model group) and the Akkermansia muciniphila intervention group (AKK group). After one week of adaptation to the environment, the Akkermansia muciniphila suspension prepared in Example 1 was gavaged once a day, with a gavage dose of 1×10 9 CFU / mL / kg / d, observed for one week, and recorded the mortality rate and weight changes of the rats. The results showed that the weight of the rats increased slightly within one week, no obvious symptoms of poisoning occurred, behavioral activities were normal, there was no death, and no adverse reactions. Therefore, it can be considered that Akkermansia muciniphila is highly safe and belongs to the normal non-toxic probiotic category.

[0044] Then, the rats in the Control group were gavaged with an equal volume of 0.5% sodium carboxymethyl cellulose (CMC-Na) at a dose of 10 mL / kg / d for 28 consecutive days. The rats in the other two groups were gavaged with an equal volume of letrozole and fed an equal amount of high-sugar and high-fat diet at a dose of 1 mg / kg / d for 28 consecutive days to induce the PCOS rat model. The three groups of rats were kept under the conditions of room temperature of 22±2℃, relative humidity of 55±5%, and light / dark alternation of 12h / 12h.

[0045] Modeling standard judgment

[0046] The criteria for successful modeling are: estrous cycle disorder and elevated testosterone T in rats.

[0047] The content of sex hormone testosterone T in the serum of rats in the Control group and the Model group was determined. Figure 1 As shown in the figure, the content of testosterone T in the serum of rats in the Model group was significantly higher than that in the Control group, indicating that the model was successfully established.

[0048] For the estrous cycle, vaginal smears were performed on rats in each group at 10 a.m. every day from the 21st day to the 28th day after letrozole and high-sugar and high-fat diet induction. The cells smeared on the slide were fixed with ethanol, stained with hematoxylin staining solution, and then the main types of cells were observed under a microscope to determine the stage of the estrous cycle of the rats.

[0049] The estrus cycle of normal rats is divided into four stages: proestrus, estrus, metestrus and diestrus. The estrus cycle shows regular changes between proestrus, estrus, metestrus and diestrus. Figure 2As shown, the cell structure of the vaginal smear of rats during different estrus cycles, among which, Figure a is the diestrus period, and the field of view is basically white blood cells; Figure b is the proestrus period, and a large number of nucleated epithelial cells can be seen, occasionally keratinized cells, and no white blood cells; Figure c is the estrus period, which is almost entirely anucleated keratinized cells, with no white blood cells and nucleated epithelial cells; Figure d is the metestrus period, with the largest number of cells in the field of view, and the amounts of the three types of cells are relatively uniform.

[0050] like Figure 3 As shown, the representative estrous cycle change diagram of the Control group and the Model group. Compared with the Control group, the estrous cycle of the Model group was mostly in the late estrus and diestrus, and the estrous cycle was disordered, which means that the model was successfully established and PCOS rats were obtained, among which P represents proestrus, E represents estrus, M represents metestrus, and D represents diestrus.

[0051] Akkermansia muciniphila intervention

[0052] After the modeling, the rats in the Control group and the Model group were given 10 mL / kg / d of distilled water by intragastric administration once a day for 21 days; the rats in the AKK group were given 1×10 9 CFU / mL / kg / d Akkermansia muciniphila suspension was administered orally once a day for 21 days. At the beginning of oral administration, the body weight of each group of rats was measured every day, and the weight change chart of each group of rats was drawn. The results are shown in Figure 2. Figure 4 As shown; during the oral gavage treatment, the rats were maintained with free access to food and clean drinking water.

[0053] from Figure 4 It can be seen that the body weight of rats in the Model group was always higher than that in the AKK group and the Control group; the body weight of rats in the AKK group showed a downward trend. It can be seen that AKK can reduce the body weight of PCOS rats.

[0054] Detection of the improving effect of AKK on PCOS rats

[0055] Fasting insulin (INS) level and glucose tolerance test

[0056] One day before the end of the intragastric administration, the rats in each group were fasted but not watered for 12 h, and then their tails were cut for blood collection. The fasting insulin (INS) content of the rats in each group was determined by enzyme-linked immunosorbent assay using an Elisa kit, and a graph showing the changes in the fasting insulin (INS) content of the rats in each group was drawn. The results are shown in Table 1. Figure 5 As shown;

[0057] from Figure 5It can be seen that, compared with the Control group and the Model group, the INS content of the rats in the Model group was significantly increased, and the difference between the two was statistically significant. Therefore, the rats in the Model group had insulin resistance, that is, after the modeling of letrozole and high-sugar and high-fat diet, the rats had insulin resistance; compared with the Model group and the AKK group, the INS content of the rats in the AKK group was significantly decreased, and the difference between the two was statistically significant. Therefore, AKK can improve the insulin resistance of PCOS rats.

[0058] Ovarian mass and ovarian morphology examination

[0059] After 21 days of drug treatment, the rats in each group were fasted for 12 hours. All rats in each group were anesthetized by inhalation of isoflurane. After successful anesthesia, they were locally disinfected with medical ethanol, the abdominal cavity was opened, and 5 mL of blood was collected through the abdominal aorta. The blood was centrifuged at 4000 r / min for 15 minutes, and the supernatant was divided into centrifuge tubes and then frozen in a -80℃ refrigerator for later use.

[0060] The ovaries of the rats were separated and washed with saline and dried with absorbent paper. The weight of the ovaries was weighed and the changes in the weight of the ovaries of the rats in each group were plotted. The results are shown in Figure 6 As shown;

[0061] The ovaries of rats in each group were fixed in 4% paraformaldehyde solution. After 24 hours, they were embedded in paraffin and stained with hematoxylin-eosin (HE). The sections were observed under an inverted microscope. The sections of the ovarian tissues of rats in each group are shown in the figure below. Figure 7 As shown;

[0062] from Figure 6 It can be seen that 21 days after administration, the ovarian mass of rats in the Model group was significantly increased compared with the Control group and the Model group, and the difference was statistically significant; the ovarian mass of rats in the AKK group was significantly decreased compared with the Model group and the AKK group, and the difference was statistically significant. It can be seen that AKK can alleviate the increase in ovarian mass of PCOS rats caused by letrozole combined with high-sugar and high-fat feed, and promote the recovery of ovarian mass of PCOS rats to normal levels.

[0063] from Figure 7 It can be seen that, comparing the Control group and the Model group, there are multiple corpora lutea in the ovaries of the rats in the Control group, which are neatly arranged and complete in morphology, and very few cystic dilated follicles are seen, while the corpora lutea in the ovaries of the rats in the Model group are smaller in size and fewer in number, and cystic dilated follicles and atretic follicles are more common, and the granulosa layer is disordered and the number of layers is reduced; comparing the Model group and the AKK group, the number of corpora lutea in the ovaries of the rats in the AKK group is increased, and cystic dilated follicles are rare; thus, it can be seen that AKK can effectively improve the ovarian lesions of PCOS rats.

[0064] Liver tissue morphology

[0065] The liver tissues of rats in each group were fixed in 4% paraformaldehyde solution. After 24 hours, the liver tissues were embedded in paraffin and stained with hematoxylin-eosin (HE). The sections were observed under an inverted microscope. Figure 8 As shown;

[0066] from Figure 8 It can be seen that the liver cell nuclei of the rats in the Control group are concentrated, round, and uniform in size, and the hepatic cords are neatly arranged; the liver cell cytoplasm of the rats in the Model group has diffuse lipid droplets of varying sizes, and some nuclei are squeezed to one side by cytoplasmic lipids, and the nuclei are deformed, displaced, and some even disappear; compared with the Model group, the lipid droplets in the AKK group are smaller and the number of vacuoles is reduced. It can be seen that AKK treatment can protect the liver from damage related to hepatocyte fatty degeneration and cholesterolemia.

[0067] Sex hormone level testing

[0068] The serum supernatant of the rat's aorta was taken and treated with an Elisa kit. Then, the absorbance of the serum supernatant of each group of rats at 450 nm was detected by an ELISA reader. The follicle stimulating hormone (FSH) content in the rats was calculated according to the software of the Elisa kit. The results are as follows: Fig. 9 As shown;

[0069] from Fig. 9 It can be seen that, comparing the Control group and the Model group, the FSH level of the rats in the Model group was significantly reduced. Comparing the Control group, the Model group and the AKK group, the FSH level of the rats in the AKK group was significantly higher than that in the Model group, and was basically the same as the FSH level of the rats in the Control group, indicating that AKK can effectively increase the content of follicle-stimulating hormone in PCOS rats to normal levels. Example 3

[0070] The water extraction and alcohol precipitation method is used to extract Tripterygium wilfordii polysaccharide. The specific steps are as follows:

[0071] The Leigongji was repeatedly washed with distilled water, dried and crushed, and then sieved through 40 mesh; then precipitated with 95% ethanol overnight to remove fat-soluble substances; after the residue was completely dried, hot water extraction was carried out at 75°C with a solid-liquid ratio of 1:50, repeated three times, the supernatant was combined, and evaporated and concentrated to a viscous state; the concentrate was precipitated with 75% ethanol overnight to collect the polysaccharide precipitate; the polysaccharide precipitate was dissolved with distilled water, and the concentrated polysaccharide solution was added with Sevag solution (chloroform: n-butanol = 4:1) at a volume of 4:1, and oscillated and mixed to remove protein, 30 minutes / time, repeated three times; then dialyzed with distilled water for 24 hours using a dialysis bag of 8000~14000Da, and the water was changed every three hours to remove small molecules such as monosaccharides to obtain Leigongji polysaccharide, which was stored at 4°C for later use.

[0072] A sugar-free MRS culture medium was prepared as a blank control group (CTRL), and a sugar-free MRS culture medium supplemented with 2% by weight of Trinita polysaccharide was prepared as a Trinita polysaccharide group (NCVP). Three parallel samples were set for each group. Each sample was inoculated with an equal amount of the Akkermansia muciniphila suspension prepared in Example 1, and the absorbance at a wavelength of 600 nm was measured every 6 hours, wherein the inoculation amount of the Akkermansia muciniphila suspension was 5 ml.

[0073] The experimental results of this embodiment are as follows Fig.10 As shown, the experimental results indicate that Tripterygium wilfordii polysaccharide can promote the growth of Akkermansia muciniphila, and can still promote Akkermansia muciniphila after 24 hours, so that Akkermansia muciniphila can achieve a long-term therapeutic effect on polycystic ovary syndrome and can solve the problem that oral Akkermansia muciniphila will be excreted from the body with feces. Example 4

[0074] This embodiment provides a composition for preventing and treating polycystic ovary syndrome, comprising the following active components: Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension; the weight ratio of Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension is 1:0.8:0.8:0.8:1.

[0075] The preparation method of the above composition comprises the following steps:

[0076] S1. Extracting Tripterygium wilfordii polysaccharide and Dendrobium tassel polysaccharide from Tripterygium wilfordii and Dendrobium tassel polysaccharide respectively by water extraction and alcohol precipitation method, and separating and purifying Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide from Hericium erinaceus mycelium crude extract and Lycium barbarum extract respectively by AKTA protein purification method, wherein the water extraction and alcohol precipitation method and the AKTA protein purification method are both prior arts;

[0077] S2. Take the Akkermansia muciniphila suspension prepared in Example 1, add the Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide thereto in sequence, and mix well. Example 5

[0078] This embodiment provides a composition for preventing and treating polycystic ovary syndrome, comprising the following active components: Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension; the weight ratio of Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide, Lycium barbarum polysaccharide and Akkermansia mucinotropis bacterial suspension is 1.1:0.8:0.8:0.8:1.1. Example 6

[0079] Eighteen SPF female SD rats aged 6 weeks were selected, with a weight of 160-180 g. The 18 rats were randomly divided into three groups, with 6 rats in each group, namely the blank group, the Akkermansia muciniphila treatment group (control group 1) and the combination treatment group (control group 2). The blank group rats were intragastrically gavaged with 0.5% sodium carboxymethyl cellulose (CMC-Na) in an equal volume every day, with an intragastrically dose of 10 mL / kg / d, for 28 consecutive days. The other two groups of rats were intragastrically gavaged with an equal volume of letrozole every day and fed with an equal amount of high-sugar and high-fat feed, with an intragastrically dose of 1 mg / kg / d, for 28 consecutive days. The PCOS rat model was induced. The three groups of rats were kept under the conditions of room temperature of 22±2℃, relative humidity of 55±5%, and light-dark alternation of 12h / 12h.

[0080] After the modeling was completed, the rats in the blank group were given 10 mL / kg / d of distilled water by gavage, the rats in the control group 1 were given 5 mL / kg / d of Akkermansia muciniphila suspension prepared in Example 1 by gavage, and the rats in the control group 2 were given 5 ml / kg / d of the composition prepared in Example 4 by gavage, once a day, for 21 days with equal volume. At the beginning of the gavage treatment, the body weight of the rats in each group was measured every 3 days, and the weight change graph of the rats in each group was drawn. The results are shown in FIG. Fig.11 As shown; during the oral gavage treatment, the rats were maintained with free access to food and clean drinking water.

[0081] Depend on Fig.11 It can be seen that after treatment, the body weight of the rats in the comparison group 2 was closer to that of the blank group, that is, the PCOS rats treated with the composition of the present invention were closer to the body weight of normal rats, indicating that the composition of the present invention has a better effect on regulating the body weight of PCOS rats than the Akkermansia muciniphila suspension.

[0082] By detecting the fasting insulin (INS) level and the sex hormone level, it was verified that the composition of the present invention has a better effect in treating polycystic ovary syndrome than the Akkermansia muciniphila suspension. The specific method is the same as that in Example 2.

[0083] The changes in fasting insulin (INS) levels of the three groups of rats are shown in the figure Fig.12 As shown by Fig.12 It can be seen that the INS content of the rats in the control group 2 is closer to the INS content of the rats in the blank group than that of the control group 1, indicating that the composition of the present invention has a better regulating effect on the fasting insulin of PCOS rats than the Akkermansia muciniphila suspension.

[0084] The results of sex hormone level test of the three groups of rats are as follows Fig.13 As shown by Fig.13 It can be seen that the FSH levels of the rats in the control group 1 and the control group 2 were basically the same as those in the blank group, indicating that the combination of the present invention and the Akkermansia muciniphila suspension had similar effects on regulating the follicle-stimulating hormone content in PCOS rats.

[0085] In summary, the present invention uses a female SD rat model induced by letrozole and a high-sugar and high-fat diet to demonstrate the application of Akkermansia muciniphila in treating polycystic ovary syndrome. The test method is safe and reliable, the test results are accurate, and the therapeutic effect is significant. The present invention provides a composition capable of inhibiting, improving or treating polycystic ovary syndrome, which has a better effect than oral administration of Akkermansia muciniphila suspension alone, and the components of the composition of the present invention are safer, which solves the technical problem that existing chemical synthetic drugs have side effects in treating polycystic ovary syndrome, and has good application prospects.

[0086] The specific implementation of the present invention described above does not constitute a limitation on the protection scope of the present invention. Any other corresponding changes and modifications made based on the technical concept of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A composition for preventing and treating polycystic ovary syndrome, characterized in that: The invention comprises the following active components: polysaccharide of Tripterygium wilfordii, polysaccharide of Dendrobium tassels, polysaccharide of Hericium erinaceus, polysaccharide of Lycium barbarum and suspension of Akkermansia muciniphila, wherein the weight ratio of polysaccharide of Tripterygium wilfordii, polysaccharide of Dendrobium tassels, polysaccharide of Hericium erinaceus, polysaccharide of Lycium barbarum and suspension of Akkermansia muciniphila is 1-1.1:0.8:0.8:0.8:1-1.1; The Akkermansia muciniphila suspension contains Akkermansia muciniphila, and the strain number of Akkermansia muciniphila in the China Industrial Microbiological Culture Collection Center CICC is CICC24917; The concentration of Akkermansia muciniphila in the Akkermansia muciniphila suspension is 1×10 9 CFU / mL; The culture method of the Akkermansia muciniphila suspension is as follows: 0.4 g of mucin and 1 mg of L-cysteine ​​are added to 200 mL of brain infusion medium BHI, and the mixture is cultured under anaerobic conditions. Then, the activated Akkermansia muciniphila is added, and the mixture is cultured at 37° C. for 24 to 48 hours. The bacterial cells are collected by centrifugation, and the bacterial cells are resuspended in distilled water to prepare a suspension with a concentration of 1×10 9 CFU / mL of Akkermansia muciniphila suspension, stored at 4°C, wherein the concentration of mucin is 2 g / L, and the concentration of L-cysteine ​​is 5 mg / L; The preparation method of the composition comprises the following steps: S1. Extracting Tripterygium wilfordii polysaccharide and Dendrobium tassellensis polysaccharide from Tripterygium wilfordii and Dendrobium tassellensis respectively by water extraction and alcohol precipitation method, and separating and purifying Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide from Hericium erinaceus mycelium crude extract and Lycium barbarum extract respectively by AKTA protein purification method; S2. Add the Tripterygium wilfordii polysaccharide, Dendrobium fringed polysaccharide, Hericium erinaceus polysaccharide and Lycium barbarum polysaccharide to the Akkermansia muciniphila suspension in sequence and mix well.

2. Use of the composition for preventing and treating polycystic ovary syndrome according to claim 1 in preparing a drug for preventing and treating polycystic ovary syndrome.

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