Application of lactobacillus acidophilus JYLA-126 in preparation of auxiliary medicine for treating polycystic ovarian syndrome by metformin

Abstract

The invention relates to the technical field of microorganisms, in particular to application of lactobacillus acidophilus JYLA-126 in preparation of auxiliary drugs for treating polycystic ovarian syndrome by metformin. According to the invention, the lactobacillus acidophilus JYLA-126 and the metformin are combined to treat the polycystic ovarian syndrome, so that the polycystic ovarian syndrome is relieved, and meanwhile, the problem of side effects of gastrointestinal tracts is reduced.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to application of Lactobacillus acidophilus JYLA-126 in preparing an auxiliary medicine for treating polycystic ovary syndrome with metformin. Background Art

[0002] Polycystic ovary syndrome (PCOS) is the most common and highly heterogeneous reproductive endocrine disease in women of childbearing age. It is also the main cause of anovulatory infertility. The incidence of PCOS is as high as 5%-20% in different populations. The basic clinical characteristics of PCOS include excessive androgen, abnormal ovulatory function and polycystic ovarian changes, accompanied by endocrine and metabolic abnormalities, including elevated luteinizing hormone (LH), insulin resistance (IR), lipid metabolism disorders and obesity. PCOS patients have a significantly higher risk of impaired glucose tolerance and type 2 diabetes, hypertension, hyperlipidemia, metabolic syndrome, cardiovascular disease, anxiety and depression than healthy women.

[0003] Insulin resistance (IR) is clinically defined as the inability of a known amount of exogenous or endogenous insulin to increase an individual's glucose uptake and utilization as in the normal population. In the state of insulin resistance, peripheral tissues (muscle, fat, liver, etc.) are less sensitive to the effects of insulin, and the efficiency of glucose uptake and utilization is reduced. Therefore, the pancreas needs to secrete higher than normal concentrations of hormones to maintain normal blood sugar, leading to the occurrence of compensatory hyperinsulinemia. Although insulin resistance is not always present in PCOS, it plays a central role in the pathogenesis of PCOS and is common in PCOS patients. The incidence of insulin resistance in PCOS patients is as high as 20%-40%, of which up to 95% of obese PCOS patients have insulin resistance. Obesity is not only a common feature of PCOS, but also an important driving factor in its pathological mechanism. Metformin can alleviate the core pathophysiological problems of PCOS through multiple mechanisms such as improving insulin resistance, regulating metabolic disorders, and inhibiting ovarian androgen production, and plays an important role in the treatment of PCOS. However, the use of metformin to treat PCOS can cause gastrointestinal side effects. Summary of the invention

[0004] In view of the problem of gastrointestinal side effects when using metformin to treat PCOS, the present invention provides an application of Lactobacillus acidophilus JYLA-126 in the preparation of an auxiliary drug for metformin to treat polycystic ovary syndrome. By combining Lactobacillus acidophilus JYLA-126 and metformin to treat polycystic ovary syndrome, the problem of gastrointestinal side effects is reduced while alleviating polycystic ovary syndrome.

[0005] The technical solution of the present invention is as follows: A use of Lactobacillus acidophilus JYLA-126 in the preparation of an auxiliary drug for metformin in the treatment of polycystic ovary syndrome. Lactobacillus acidophilus JYLA-126 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on July 8, 2019, with the deposit address being No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number being CGMCC No. 18095.

[0006] Further, the functions of adjuvant drugs include treating polycystic ovary syndrome, alleviating gastrointestinal adverse reactions and / or treating insulin resistance.

[0007] Furthermore, PCOS is treated by increasing anti-Mullerian hormone levels and decreasing luteinizing hormone levels.

[0008] Furthermore, polycystic ovary syndrome and / or insulin resistance can be treated by regulating the relevant intestinal flora.

[0009] Furthermore, polycystic ovary syndrome and / or insulin resistance can be treated by increasing the abundance of relevant intestinal flora.

[0010] Furthermore, the relevant intestinal flora included Firmicutes ( Firmicutes ) and Bacteroidetes ( Bacteroidetes ) flora.

[0011] Furthermore, gastrointestinal adverse reactions include one or more of upper abdominal pain, acid reflux, nausea, abdominal distension, belching, constipation and diarrhea. Lactobacillus acidophilus JYLA-126 can significantly reduce upper abdominal pain, acid reflux, nausea, and especially significantly improve diarrhea.

[0012] Furthermore, gastrointestinal adverse reactions are gastrointestinal adverse reactions caused by metformin treatment in patients with polycystic ovary syndrome with insulin resistance and obesity. Lactobacillus acidophilus JYLA-126 can regulate the intestinal flora to alleviate the gastrointestinal adverse reactions caused by metformin treatment in patients with polycystic ovary syndrome with insulin resistance and obesity. Regulating the intestinal flora includes improving the uniformity of the intestinal microbial community and regulating the composition of the intestinal flora. The intestinal flora includes Firmicutes ( Firmicutes ) flora, Bacteroidetes ( Bacteroidetes ) and Actinobacteria ( Actinobacteria Improving the uniformity of intestinal microbial communities can improve the intestinal environment, repair the intestinal mucosal barrier, regulate the intestinal immune system, promote the absorption of nutrients and stimulate intestinal peristalsis, which is beneficial to improving metabolic health. Regulating intestinal composition includes increasing the relative abundance of beneficial bacteria and reducing the relative abundance of bacteria associated with inflammation.

[0013] Furthermore, insulin resistance is treated by reducing fasting blood glucose concentration, postprandial blood glucose concentration and insulin resistance index. The insulin resistance index (HOMA-IR) is calculated according to the following formula: Fasting blood glucose level (FPG, mmol / L) × fasting insulin level (FINS, μU / mL) / 22.5.

[0014] The insulin resistance index (HOMA-IR) of a normal individual is 1. When the insulin resistance level increases, the insulin resistance index (HOMA-IR) will increase to greater than 1. Lactobacillus acidophilus JYLA-126 can not only reduce fasting blood sugar levels, but also reduce postprandial blood sugar levels.

[0015] Furthermore, the functions of auxiliary drugs also include improving reproductive outcomes. Lactobacillus acidophilus JYLA-126 can significantly improve pregnancy scores. Lactobacillus acidophilus JYLA-126 can also improve follicle-stimulating hormone (FSH) levels, testosterone levels, and ovulation scores.

[0016] The beneficial effects of the present invention are: The invention provides an application of Lactobacillus acidophilus JYLA-126 in preparing an auxiliary drug for treating polycystic ovary syndrome with metformin. Only a single strain of Lactobacillus acidophilus JYLA-126 is used to assist metformin in treating polycystic ovary syndrome with insulin resistance and obesity. On the one hand, the safety and stability of the auxiliary drug are improved, making the auxiliary drug more suitable for large-scale and batch production; on the other hand, the symptoms of polycystic ovary syndrome can be alleviated, the insulin resistance level of obese patients with polycystic ovary syndrome can be reduced, and the gastrointestinal adverse reactions caused by metformin can be alleviated. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] 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 or the description of the prior art will be briefly introduced below. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0018] Figure 1 It is the index scatter plot of patients in Group E and Group Y in Example 2 before treatment, one month after treatment, two months after treatment, and three months after treatment. Among them, A is a scatter plot of anti-Mullerian hormone level, B is a scatter plot of luteinizing hormone level, C is a scatter plot of testosterone level, D is a scatter plot of ovulation score, E is a scatter plot of fasting blood glucose level, F is a scatter plot of blood glucose level 2 hours after meal, G is a scatter plot of fasting insulin level, H is a scatter plot of insulin resistance index, *P<0.05, **P<0.01, ***P<0.001.

[0019] Figure 2 It is the intestinal microbial diversity test diagram of patients in group E and group Y in Example 3 before treatment and three months after treatment. Among them, A1 is the Chao1 diversity index diagram, B1 is the Shannon diversity index diagram, C1 is the Venn diagram, and D1 is the Beta diversity analysis diagram. In A1, B1, C1 and D1, C group represents the intestinal microbial community composition of patients in group E and group Y before treatment, N=40 (samples were randomly selected from the included participants); E group represents the intestinal microbial community composition of patients in group E after 3 months of treatment, N=40; Y group represents the intestinal microbial community composition of patients in group Y after 3 months of treatment, N=40. *P<0.05.

[0020] Figure 3 It is a test chart of the intestinal microbial composition of patients in group E and group Y in Example 4 before treatment and three months after treatment. Among them, A2 is the relative abundance of the microbiome at the phylum level, B2 is the relative abundance of the microbiome at the genus level, C2 is the relative abundance of the Firmicutes, D2 is the relative abundance of the Actinobacteria, E2 is the relative abundance of the genus Faecalibacterium, F2 is the relative abundance of the genus Eubacterium, G2 is the relative abundance of the genus Collinsella, and H2 is the relative abundance of the genus Ruminococcus. In A2-H2, C group represents the intestinal microbial composition of patients in group E and group Y before treatment, N=40 (samples were randomly selected from the included participants); E group represents the intestinal microbial composition of patients in group E after 3 months of treatment, N=40; Y group represents the intestinal microbial composition of patients in group Y after 3 months of treatment, N=40. *P<0.05, **P<0.01. The relative abundance of these taxa between groups was compared using one-way ANOVA, followed by Tukey's post hoc test when the assumption of variance homogeneity was met. In cases where variance homogeneity was not met, Tamhane's T2 method was used for post hoc test and P value correction.

[0021] Figure 4 is the score of gastrointestinal adverse reactions in Group E and Group Y in Example 5. In A3-D3, A3 is the score of upper abdominal pain, B3 is the score of acid reflux, C3 is the score of nausea, and D3 is the score of diarrhea. group represents the score of gastrointestinal adverse reactions of patients in Group E after 3 months of treatment; group represents the score of gastrointestinal adverse reactions of patients in Group Y after 3 months of treatment. *P<0.05, **P<0.01.

[0022] Figure 5 This is a correlation analysis chart of the microbial profile in Example 6 with PCOS and insulin resistance. DETAILED DESCRIPTION

[0023] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0024] Example 1 Clinical patient recruitment and baseline characteristics One hundred obese patients with polycystic ovary syndrome and insulin resistance were randomly recruited as research subjects and divided into two groups, named Group E and Group Y. In addition, 50 patients before treatment were randomly selected from the enrolled patients as the control group, named Group C. Patients in Group E received metformin + placebo treatment, and the medication conditions were as follows: each patient took 0.5g of metformin tablets and 2g of placebo each time, starting from the third day of the menstrual cycle, and took metformin and placebo three times a day; patients in Group Y received metformin + Lactobacillus acidophilus JYLA-126 combined treatment, and the medication conditions were as follows: each patient took 0.5g of metformin tablets and 2g of Lactobacillus acidophilus JYLA-126 freeze-dried powder each time, and the number of viable bacteria of Lactobacillus acidophilus JYLA-126 freeze-dried powder was 1×10 9 CFU / g, oral administration from the third day of the menstrual cycle, taking metformin and Lactobacillus acidophilus JYLA-126 freeze-dried powder three times a day. Both groups E and Y were followed up monthly, and four weeks of doses were given during the monthly follow-up for twelve consecutive weeks. During the follow-up period, one patient in group E and three patients in group Y were lost to follow-up. Therefore, the final number of patients in group E was 49, and the final number of patients in group Y was 47. The basic information and characteristics of the patients in groups E and Y are shown in Table 1. In Table 1, BMI is body mass index. Scoring criteria: Menstrual cycle regularity and pregnancy history were manually scored as binary variables, and the scores of manual scoring were 0 or 1. In the regularity of the menstrual cycle, 1 point indicates normal menstrual cycle, and 0 points indicate menstrual disorders. In the pregnancy history, 1 point indicates a history of pregnancy, and 0 points indicate no such history. Data display and statistical analysis in Table 1: Data are expressed as mean ± standard deviation. The independent sample t test was used to compare the groups, and P ≥ 0.05 indicated that there was no statistically significant difference. Lactobacillus acidophilus JYLA-126 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on July 8, 2019. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The deposit number is CGMCC No. 18095 and the classification name is Lactobacillus acidophilus Lactobacillus acidophilusThe strain has been disclosed in Chinese patent CN116159082A, and the patent name of Chinese patent CN116159082A is: a product for preventing and / or treating obesity and metabolic syndrome, and its preparation method and application.

[0025] Table 1 Basic information and characteristics of patients in group E and group Y

[0026] In Table 1 , the total number of patients refers to the sum of the number of patients in group E and the number of patients in group Y, and the totals of age, BMI, menstrual cycle regularity, and maternal and childbearing history refer to the mean values ​​plus deviations of age, BMI, menstrual cycle regularity, and maternal and childbearing history of patients in group E and group Y, respectively.

[0027] As shown in Table 1, the baseline characteristics between Group E and Group Y showed a well-matched distribution, ensuring comparable starting points for evaluating the effect of Lactobacillus acidophilus JYLA-126 adjunct to metformin treatment on study outcomes. Key parameters such as age, BMI, menstrual cycle regularity, and maternal history were similar between the two groups, facilitating robust comparisons.

[0028] The case diagnosis criteria of the research subjects are as follows: (1) Diagnostic criteria for PCOS patients (using the internationally recognized Rotterdam criteria): ① Oligomenorrhea or amenorrhea or irregular uterine bleeding.

[0029] ② There is clinical and / or biochemical evidence of androgen excess.

[0030] ③Ultrasound examination showed polycystic ovaries. The number of follicles with a diameter of 2mm-9mm in one or both ovaries was ≥12, and (or) the ovarian volume was ≥10ml.

[0031] The diagnosis can only be made if two of the above three conditions are met and eliminated. Other causes of hyperandrogenism such as congenital adrenal hyperplasia, tumors that secrete androgens, Cushing's syndrome, etc. are excluded; other diseases that cause ovulation disorders such as hyperprolactinemia, premature ovarian dysfunction, hypogonadotropic amenorrhea, and thyroid dysfunction are excluded.

[0032] (2) The diagnostic criteria for obesity are based on the "Chinese Expert Consensus on Medical Nutritional Treatment for Overweight / Obesity (2016 Edition)", which states that 24≤BMI<28kg / m² is considered overweight, and BMI≥28kg / m² is considered obese.

[0033] (3) The diagnostic criteria for insulin resistance are based on the Expert Guidance on Methods and Applications of Insulin Resistance Assessment (2018 Edition): The formula of the HOMA-IR steady-state model (a simple assessment of insulin resistance that is easy to operate and low-cost) is HOMA-IR = fasting blood glucose level (FPG, mmol / L) × fasting insulin level (FINS, μU / mL) / 22.5. The HOMA-IR index of a normal individual is 1. As the level of insulin resistance increases, the HOMA-IR index will be higher than 1, and a value exceeding 2.69 is considered insulin resistance.

[0034] The inclusion criteria for the study subjects were as follows: (1) Those who meet the diagnostic criteria for polycystic ovary syndrome combined with obesity and insulin resistance; (2) Women of childbearing age, aged 18-40 years; (3) Not taking any medications that affect menstruation (such as contraceptives, hormones, or other medications that affect insulin, blood sugar, sex hormones, and blood lipid metabolism) or using any related treatments within three months, and no record of artificial abortion; (4) Gynecological examination and auxiliary examinations: no organic lesions of the internal and external reproductive organs, and no other primary diseases; (5) Those who have signed the informed consent form.

[0035] The exclusion criteria for the study subjects were as follows: (1) Combined with other endocrine diseases, such as diabetes, thyroid disease, hormone-secreting tumors, etc.; (2) Patients with severe or unstable physical illnesses, including liver, kidney, cardiovascular, respiratory, nervous or blood system diseases, autoimmune diseases, and mental illnesses; (3) having a history of thromboembolic disease or thrombosis tendency; (4) Women who are heavy smokers or alcoholics; (5) Women who are breastfeeding or pregnant; (6) Women with a history of other malignant tumors; (7) Use of statins / antibiotics / probiotic products / traditional Chinese medicine or other drugs known or suspected to affect reproductive or metabolic function within the past 3 months.

[0036] (8) Special physical condition, allergic to drugs; (9) Patients who need other treatments besides this disease, which may affect this study and cannot be carried out according to this study plan; (10) Patients with congenital deficiencies in the reproductive system who cannot meet the requirements of this study.

[0037] Example 2 Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in improving insulin resistance and alleviating symptoms of polycystic ovary syndrome 1. Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in alleviating symptoms of polycystic ovary syndrome (1) Blood samples were collected from patients in group E and group Y before treatment, one month after treatment, two months after treatment, and three months after treatment. The blood samples were collected from patients on the 2nd to 4th day of menstrual bleeding, between 8:00 and 9:00 a.m. The blood samples were obtained by drawing fasting peripheral venous blood from the patients, centrifuging it, and separating the serum.

[0038] After the blood samples were collected, the levels of anti-Mullerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (TEST) of the patients were measured using a fully automatic electrochemiluminescence immunoassay. The test results are shown in Table 2.

[0039] (2) Evaluation of ovulation score and pregnancy score The ovulation and pregnancy status of patients who wanted to prepare for pregnancy at the time of enrollment were followed up, and the average ovulation score and the average pregnancy score of patients in group E and group Y were evaluated by binary scoring method. The average ovulation score and the average pregnancy score are shown in Table 2, where 0 = no ovulation or no pregnancy, and 1 = ovulation or pregnancy.

[0040] (II) Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in improving insulin resistance The fasting blood glucose (FPG), 1-hour postprandial blood glucose (1h-PBG), and 2-hour postprandial blood glucose (2h-PBG) levels of the above blood samples were detected by standard automated detection methods (Beckmen AU series fully automated chemical analyzer), and the serum fasting insulin levels (FINS) of the above blood samples were determined by chemiluminescent immunoassay (Siemens Centaur XP). The test results are shown in Table 2. Insulin sensitivity and pancreatic β-cell function were evaluated using several mathematical models or indicators.

[0041] Among them, the homeostatic model of insulin resistance (HOMA-IR): HOMA-IR index = fasting blood glucose level (FPG, mmol / L) × fasting insulin level (FINS, μU / mL) / 22.5; The HOMA-IR index of a normal individual is 1. As the level of insulin resistance increases, the HOMA-IR index will be higher than 1. The calculation results of the HOMA-IR index of Group E and Group Y are shown in Table 2.

[0042] Table 2. Blood sample test results of Group E and Group Y

[0043] Abbreviations and definitions: 0M: baseline (before treatment); 1M: after 1 month of treatment; 3M: after 3 months of treatment. G*T: group*time interaction. AMH: anti-Mullerian hormone; FSH: follicle-stimulating hormone; LH: luteinizing hormone; TEST: testosterone; FPG: fasting plasma glucose; 1h-PBG: 1-hour postprandial plasma glucose; 2h-PBG: 2-hour postprandial plasma glucose; HOMA-IR: homeostasis model assessment of insulin resistance, calculated as HOMA-IR index=FPG×FPINS / 22.5.

[0044] Statistical analysis: Data are presented as mean ± standard deviation (SD). Two-way ANOVA and Bonferroni post hoc test were used to analyze the differences between groups at different time periods, and the P values ​​were adjusted. Independent sample t-test was used to evaluate the statistical significance of pregnancy rates between groups. *P<0.05, **P<0.01, ***P<0.001, NS: not significant.

[0045] Table 2 provides a comprehensive comparison of clinical parameters between Group E and Group Y before treatment (0M), one month after treatment (1M), and three months after treatment (3M), including hormone indicators and HOMA-IR index. As shown in Table 2, patients in Group Y who received adjuvant treatment with Lactobacillus acidophilus JYLA-126 showed significant improvements in multiple PCOS-related symptom parameters. Specifically, Figure 1 As shown, the anti-Mullerian hormone (AMH) level in group Y increased from 10.43±3.56 ng / mL at baseline to 6.40±1.88 ng / mL after three months (p<0.05). The luteinizing hormone (LH) level in group Y also showed significant improvement, decreasing from 11.66±2.79 U / L at baseline to 7.68±1.15 U / L after three months (p<0.05 at 3 months; p<0.01 at 1 month). Although the follicle-stimulating hormone (FSH) and testosterone (TEST) levels and ovulation scores within group Y also improved significantly, these changes did not reach statistical significance between the two groups. Notably, the mean pregnancy score of group Y was 0.70±0.46, which was significantly higher than the mean pregnancy score of group E, 0.49±0.51, p=0.003, indicating that the fertility outcome of the adjuvant treatment group with Lactobacillus acidophilus JYLA-126 was enhanced.

[0046] When evaluating insulin resistance markers after one month of treatment, group Y was significantly lower than group E in several key indicators: the fasting blood glucose (FPG) concentration of group Y was 4.97±0.53mmol / L, which was lower than the fasting blood glucose (FPG) concentration of group E (5.53±0.84mmol / L) (p<0.001), the 2-hour postprandial blood glucose (2h-PBG) level of group Y was 6.75±0.60mmol / L, which was lower than the 7.52±0.86 mmol / L of group E (p<0.001), and the HOMA-IR index of group Y was 3.35±1.48, which was lower than the 4.40±2.29 of group E (p<0.05). Although both groups E and Y showed significant improvements in fasting insulin (FINS) and 1-hour postprandial blood glucose (1h-PBG) levels, these changes did not reach statistical significance between the groups. In addition, after three months, both groups E and Y showed significant improvements in all IR parameters relative to pre-treatment, but no significant differences were observed between the groups. These findings highlight the potential advantages of Lactobacillus acidophilus JYLA-126 as an adjunct to metformin in improving markers of insulin resistance. From the above, it can be seen that the addition of Lactobacillus acidophilus JYLA-126 can not only improve the clinical symptoms of PCOS, but also improve metabolic health, enhancing its benefits in the management of PCOS.

[0047] Example 3 Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in improving the uniformity of intestinal microbial communities Fecal samples were collected from patients in group E and group Y before treatment and three months after treatment. The time of collecting fecal samples was: 2-4 days of menstrual bleeding. The method of obtaining fecal samples was as follows: a) prepare the bedpan and fecal container, wash hands, and wear gloves to collect fresh fecal samples; b) immediately package and label the fecal samples of the subjects in the laboratory; c) use a sterile toothpick or fecal sampler to cut the middle part of the sample (the surface of the feces contains exfoliated cells of the intestinal mucosa; the outside is easily contaminated, and after contact with the air, some bacterial DNA begins to degrade), take about 0.5-1.5g (about the size of a peanut) into a sterile 2.0mL centrifuge tube, and take 3-5 tubes for each sample as backup; d) After packaging, immediately freeze in liquid nitrogen or directly store in a low-temperature environment of -80℃. When sending samples, choose dry ice transportation. If the fecal sample is large or cannot be frozen immediately, it must be collected within 2 hours at the latest.

[0048] After collecting stool samples, 6SrRNA high-throughput sequencing (at least 30 stool samples per group) and non-targeted metabolomics testing were performed on the stool samples. The test results were as follows: Figure 2 shown.

[0049] Depend on Figure 2Figure A1 shows that the Chao 1 index of group C patients was 249.67, the Chao 1 index of group E patients was 254.087, and the Chao 1 index of group Y patients was 275.491. Compared with group E, the Chao 1 index of group Y patients did not change much, indicating that the species richness of the intestinal microecology of patients remained stable before and after the use of Lactobacillus acidophilus JYLA-126 to assist metformin treatment, which indicates that adjuvant therapy did not significantly change the diversity of different microbial groups. Figure 2 Figure B1 shows that the Shannon index of patients in group C was 5.3106, the Shannon index of patients in group E was 5.24475, and the Shannon index of patients in group Y was 5.54938. After the use of Lactobacillus acidophilus JYLA-126 as an adjuvant to metformin treatment, the Shannon index of patients in group Y was significantly higher than that of group E and the baseline measurement value (p<0.05). This finding suggests that the use of Lactobacillus acidophilus JYLA-126 as an adjuvant treatment may enhance the uniformity of the microbial community. Figure 2 As shown in Figure C1, the Venn diagram analysis identified a core set of 679 operational taxonomic units (OTUs) shared by all groups, and highlighted the differences in the number of unique OTUs between groups C, E, and Y, which were 2488, 2795, and 2938, respectively. These differences reflect the different microbial compositions within groups C, E, and Y. Figure 2 As shown in Figure D1, principal coordinate analysis (PCoA) further clarified the subtle differences in the overall microbial composition among groups C, E, and Y. Taken together, these results suggest that Lactobacillus acidophilus JYLA-126, as an adjunct to metformin, has a beneficial effect on the gut microbiota of PCOS patients and may help improve metabolic health.

[0050] Example 4 Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in regulating the composition of intestinal flora The stool samples collected in Example 3 were comprehensively analyzed. After high-throughput sequencing of the stool samples, one-way ANOVA was used to analyze whether the differences were statistically significant. The relationship between intestinal flora and key clinical indicators of PCOS was evaluated by Pearson correlation analysis. The analysis results are as follows: Figure 3 As shown, Figure 3 The analysis of changes in the intestinal flora at the phylum level and genus level in patients in group E and group Y is provided. As shown in Figure A2, at the phylum level, patients in group Y had a higher number of Firmicutes ( Firmicutes ), Bacteroidetes ( Bacteroidetes ) and Actinobacteria ( Actinobacteria) became the main microbial group. As shown in Figure B2, analysis at the genus level showed that there were significant differences between the top 15 most common genera in groups C, E, and Y. It is worth noting that, as shown in Figure C2, in group C, the proportion of Firmicutes was 51%±11%, while it increased to 56%±10% in group E and further increased to 60%±13% in group Y. This trend suggests that the addition of Lactobacillus acidophilus JYLA-126 may enhance the relative abundance of Firmicutes. As shown in Figures E2, G2, and H2, in group Y, beneficial genera such as Faecalibacterium ( Faecalibacterium ) was 10.0%±5.0% vs 7.0%±4.0% in group C, and Ruminococcus ( Ruminococcus The relative abundance of group A was 4.0%±4.0% vs 2.0%±2.0% in group B, both of which increased significantly, while the potential pro-inflammatory genus Collinsella ( Collinsella ) decreased to 5%±5%. In addition, as shown in Figures D2 and F2, the relative abundance of Actinobacteria in Group Y showed a downward trend compared with Groups C and E, while the relative abundance of Blautia ( Blautia ) showed an upward trend. These findings highlight the potential of Lactobacillus acidophilus JYLA-126 in modulating the composition of the gut microbiota in PCOS patients. By promoting beneficial genera and reducing genera associated with inflammation, this probiotic adjunctive treatment may improve metabolic health through its effects on the gut microbiome.

[0051] Example 5 Verification of the effectiveness of Lactobacillus acidophilus JYLA-126 in assisting metformin in alleviating gastrointestinal adverse reactions The Gastrointestinal Symptom Rating Scale (GSRS) was used to assess symptoms including upper abdominal pain, acid reflux, nausea, bloating, belching, constipation, and diarrhea. The results of the comparative analysis are summarized in Tables 3 and Figure 4 Table 3 highlights the effects of Lactobacillus acidophilus JYLA-126 on the above-mentioned gastrointestinal symptoms associated with metformin treatment. Figure 4As shown, key data showed that Lactobacillus acidophilus JYLA-126 adjunct to metformin in the treatment of PCOS was able to significantly reduce upper abdominal pain (mean score: Y group = 1.15 ± 0.47 vs. E group = 1.47 ± 1.00; p < 0.05), acid reflux (mean score: Y group = 1.09 ± 0.28 vs. E group = 1.43 ± 0.94; p < 0.05), nausea (mean score: Y group = 1.13 ± 0.40 vs. E group = 1.57 ± 1.21; p < 0.05), and particularly significantly improved diarrhea (mean score: Y group = 1.11 ± 0.38 vs. E group = 1.71 ± 1.41; p < 0.01). Overall, this example highlights the potential benefits of Lactobacillus acidophilus JYLA-126 as an adjunct therapy in reducing gastrointestinal adverse reactions in PCOS patients receiving metformin.

[0052] Table 3 Gastrointestinal adverse reaction scores of patients in group E and group Y

[0053] Gastrointestinal adverse reactions were evaluated using the Gastrointestinal Symptom Rating Scale (GSRS), which evaluates symptoms such as upper abdominal pain, acid reflux, nausea, and diarrhea. The GSRS uses a 7-point Likert scale, with 1 representing "no symptoms", 4 representing "moderate", and 7 representing "very severe". Data are expressed as mean ± standard deviation (SD). Statistical significance was evaluated using an independent sample t-test, with P < 0.05 indicating statistically significant differences.

[0054] Example 6 Correlation analysis between intestinal flora and insulin resistance, PCOS symptoms and other key clinical variables Pearson correlation analysis was performed to evaluate the relationship between microbial profiles and key clinical variables related to PCOS and insulin resistance. Figure 5 As shown in Figure 2, a comprehensive analysis of the significant associations between specific microbial groups and various biochemical markers. Specifically, Firmicutes ( Firmicutes ), Blautia spp. Blautia ) and Ruminococcus spp. ( Ruminococcus ) was positively correlated with FSH, ovulation rate and pregnancy rate, but negatively correlated with fasting insulin (FINS) and HOMA-IR index (p < 0.05). Bacteroidetes) were positively correlated with FINS and HOMA-IR index, but negatively correlated with anti-Mullerian hormone (AMH) and ovulation rate (p<0.05). Multiple phyla and genera showed particularly strong correlations (p<0.01 or p<0.001), highlighting their potential as therapeutic targets in patients with PCOS and insulin resistance. Notably, the abundance of Firmicutes, Blautia, and Ruminococcus was significantly higher in group Y, suggesting that Lactobacillus acidophilus JYLA-126 may promote the proliferation of these beneficial taxa, thereby potentially alleviating symptoms of PCOS and insulin resistance (p<0.05, p<0.01, or p<0.001). These results highlight the potential of Lactobacillus acidophilus JYLA-126 in the management of PCOS as an adjunctive treatment to improve insulin resistance by modulating the gut microbiota. The observed correlations suggest that this probiotic strain may play a key role in disease management by promoting a healthier microbiome, thereby enhancing metabolic health and improving reproductive outcomes in affected individuals.

[0055] Although the present invention has been described in detail with reference to the accompanying drawings and in combination with the preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and these modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily think of changes or substitutions within the technical scope disclosed by the present invention, and these shall be within the scope of protection of the present invention.

Claims

1. A use of Lactobacillus acidophilus JYLA-126 in the preparation of an auxiliary drug for metformin in the treatment of polycystic ovary syndrome.

2. The use of Lactobacillus acidophilus JYLA-126 according to claim 1 in preparing an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: Functions of adjunctive medications include treating PCOS, alleviating gastrointestinal adverse effects, and / or treating insulin resistance.

3. The use of Lactobacillus acidophilus JYLA-126 as claimed in claim 2 in the preparation of an auxiliary drug for metformin in the treatment of polycystic ovary syndrome, characterized in that: Treat PCOS by increasing anti-Mullerian hormone levels and decreasing luteinizing hormone levels.

4. The use of Lactobacillus acidophilus JYLA-126 as claimed in claim 2 in the preparation of an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: Treatment of polycystic ovary syndrome and / or insulin resistance by regulating relevant intestinal flora.

5. The use of Lactobacillus acidophilus JYLA-126 as claimed in claim 4 in the preparation of an auxiliary drug for metformin in the treatment of polycystic ovary syndrome, characterized in that: Treating polycystic ovary syndrome and / or insulin resistance by increasing the abundance of relevant gut microbiota.

6. Use of a Lactobacillus acidophilus JYLA-126 according to claim 4 in preparing an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: The relevant intestinal flora include Firmicutes ( Firmicutes ) and Bacteroidetes ( Bacteroidetes ) flora.

7. The use of Lactobacillus acidophilus JYLA-126 as claimed in claim 2 in the preparation of an auxiliary drug for metformin in the treatment of polycystic ovary syndrome, characterized in that: Gastrointestinal adverse reactions include one or more of upper abdominal pain, acid reflux, nausea, bloating, belching, constipation and diarrhea.

8. Use of a Lactobacillus acidophilus JYLA-126 according to claim 2 or 4 in the preparation of an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: Gastrointestinal adverse reactions are gastrointestinal adverse reactions caused by metformin treatment in patients with polycystic ovary syndrome with insulin resistance and obesity.

9. The use of Lactobacillus acidophilus JYLA-126 as claimed in claim 2 in the preparation of an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: Treat insulin resistance by reducing fasting blood glucose concentration, postprandial blood glucose concentration and insulin resistance index.

10. Use of Lactobacillus acidophilus JYLA-126 according to claim 1 in preparing an auxiliary drug for treating polycystic ovary syndrome with metformin, characterized in that: The function of adjunctive medicine also includes improving reproductive outcomes.

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