A traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome

Abstract

The application discloses a traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome, which comprises 13-17 parts of Baijiangxi, 7-11 parts of Huangqin, 7-11 parts of Ganjiang, 7-11 parts of Dangshen, 7-11 parts of Zhigancao, 4-8 parts of Dazao, 1-5 parts of Huanglian, 15-20 parts of Shanyao, 7-11 parts of Guchao Cangzhu, 10-15 parts of Danshen, 7-11 parts of Yiyanghe, and 8-12 parts of chrysanthemum powder. The traditional Chinese medicine composition has the remarkable effects of regulating reproductive endocrine, improving glucose metabolism, reducing blood sugar level, relieving insulin resistance symptoms, protecting ovary and promoting corpus luteum production, effectively reversing pathological damage of ovary caused by PCOS and resisting inflammation. The composition is based on the modification of Banxia Xiexin Decoction and combined with the new resource food chrysanthemum powder. Through the synergistic effect of 'traditional Chinese medicine compound regulating middle-jiao Qi and body fluid' and 'prebiotic remodeling intestinal microecology', the composition can reduce HOMA-IR index and restore autonomous ovulation function, and has important theoretical value and clinical conversion prospect for promoting the modernization of traditional Chinese medicine.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition for treating polycystic ovary syndrome (PCOS), specifically to a new resource food composition of traditional Chinese medicine for treating insulin-resistant PCOS, its preparation method and application, belonging to the field of traditional Chinese medicine composition preparation technology. Background Technology

[0002] Polycystic ovary syndrome (PCOS) is the most common complex endocrine and metabolic disorder in women of reproductive age, characterized by hyperandrogenemia, ovulation dysfunction, and polycystic ovarian morphology. Insulin resistance (IR) is its core pathological driver, hence it can be defined as insulin-resistant polycystic ovary syndrome (IR-PCOS), which is also the most prevalent clinical phenotype of PCOS. IR-PCOS is mainly caused by decreased insulin sensitivity and compensatory hyperinsulinemia, leading to endocrine disorders and metabolic imbalances, which in turn induce ovulation disorders, reproductive abnormalities, and systemic metabolic complications. It is a significant cause of high incidence of female reproductive disorders and metabolic diseases. Epidemiological data shows that the global prevalence of PCOS in women of reproductive age is 5%–18%, with 40%–70% of PCOS patients also having insulin resistance. In my country, the incidence of insulin resistance in Han Chinese PCOS patients is as high as 56.3%, far exceeding the 25% prevalence in the general female population, making it a highly prevalent chronic disease affecting women of reproductive age. The prevalence of insulin resistance (IR-PCOS) increases significantly with age, weight, and metabolic status. The incidence of insulin resistance in adolescent PCOS patients can reach 35%–80%. Obese PCOS patients have a 2–3 times higher risk of developing IR compared to non-obese patients. IR-PCOS accounts for more than 65% of all PCOS patients in adult women of childbearing age. The long-term consequences of this disease are significant, with a much higher incidence of complications than in healthy individuals. Data shows that IR-PCOS patients have a 3.45 times higher risk of developing type 2 diabetes than ordinary women, and the risk is as high as 4.8 times for obese subtype patients. The incidence of impaired glucose tolerance in these patients can reach 35%, far exceeding the 10% level in the general population. Simultaneously, the proportion of patients with hypertension reaches 45%, and the proportion with dyslipidemia reaches 60%, which are 2.25 times and 2 times higher than those in healthy women, respectively.

[0003] Current PCOS interventions mainly include lifestyle interventions, weight management, insulin sensitizers, medications to regulate menstrual cycles, and ovulation induction therapy. Some traditional Chinese medicine (TCM) formulas have multi-target and holistic regulatory characteristics, and can be used to improve menstrual irregularities, obesity, insulin resistance, and accompanying symptoms. However, existing solutions still have the following shortcomings: some drugs only treat the symptoms, not the root cause; some solutions focus on single-target blood sugar lowering, menstrual regulation, or ovulation induction, without sufficient attention to the relationship between insulin resistance (IR), gut microbiota, and reproductive endocrinology; some TCM formulas contain many ingredients, making quality control difficult; some solutions list food ingredients alongside TCM herbs without distinguishing their source, uses, and technical roles; and some solutions lack clear dosage forms, preparation processes, quality indicators, and applicable population limitations.

[0004] Recent studies have shown that PCOS patients often have altered gut microbiota structure, abnormal intestinal barrier function, and changes in metabolites. Gut microbiota dysbiosis may affect host energy metabolism, bile acid metabolism, short-chain fatty acid production, endotoxin exposure, and chronic inflammatory responses, thereby participating in the development of insulin resistance (IR). Therefore, using gut microbiota as a starting point for adjuvant intervention in PCOS patients has certain research value. Inulin is a naturally occurring β-2,1-fructan, usually extracted from plants such as Jerusalem artichoke and chicory. Inulin is not easily broken down by digestive enzymes in the stomach and small intestine, but can enter the large intestine and be selectively utilized by specific gut microbes, producing metabolites such as short-chain fatty acids. Existing literature indicates that inulin has dietary fiber and prebiotic-like effects, and can affect fecal volume, intestinal transit, and postprandial blood glucose response in some individuals. Banxia Xiexin Decoction, from the *Shanghan Lun*, is composed of Pinellia ternata, Scutellaria baicalensis, dried ginger, ginseng, Coptis chinensis, jujube, and licorice, and has the effects of balancing cold and heat, opening up and descending, harmonizing the stomach and eliminating stagnation. Modern research suggests that this formula and its modifications can be used for various chronic diseases, including gastrointestinal dysfunction, metabolic abnormalities, and digestive symptoms.

[0005] However, the existing technology has not fully disclosed a traditional Chinese medicine-food composition and its granules that rationally combines the basic structure of Banxia Xiexin Decoction, spleen-strengthening, phlegm-resolving, and blood-activating medicinal materials with inulin, a new resource food component, and is specifically used for IR-PCOS patients. Summary of the Invention

[0006] The technical problem to be solved by this invention is to address the shortcomings of existing technologies by providing a traditional Chinese medicine-new resource food composition for treating insulin-resistant polycystic ovary syndrome, its preparation method, and its application. The composition of this invention is based on a modification of Banxia Xiexin Decoction, combined with the new resource food inulin. Through the synergistic effect of "regulating the Qi mechanism of the middle Jiao with traditional Chinese medicine compound" and "reshaping the intestinal microecology with prebiotics", it achieves the effects of reducing the HOMA-IR index and restoring spontaneous ovulation. This has important theoretical value and clinical translation prospects for promoting the modernization of traditional Chinese medicine.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] One objective of this invention is to provide a traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome, comprising, by weight, the following raw materials:

[0009] Components of the prepared Chinese herbal medicine: 13-17 parts of Pinellia ternata, 7-11 parts of Scutellaria baicalensis, 7-11 parts of dried ginger, 7-11 parts of Codonopsis pilosula, 7-11 parts of prepared licorice root, 4-8 parts of jujube, 1-5 parts of Coptis chinensis, 15-20 parts of Dioscorea opposita, 7-11 parts of stir-fried Atractylodes lancea, 10-15 parts of Salvia miltiorrhiza, and 7-11 parts of Epimedium brevicornu;

[0010] New resource food component: 8-12 parts inulin.

[0011] In the above technical solution, the traditional Chinese medicine composition preferably includes the following raw materials in parts by weight: 14-16 parts of Pinellia ternata, 8-10 parts of Scutellaria baicalensis, 8-10 parts of dried ginger, 8-10 parts of Codonopsis pilosula, 8-10 parts of prepared licorice root, 5-7 parts of jujube, 2-4 parts of Coptis chinensis, 15-18 parts of Dioscorea opposita, 8-10 parts of stir-fried Atractylodes lancea, 11-14 parts of Salvia miltiorrhiza, 8-10 parts of Epimedium brevicornu, and 9-11 parts of inulin.

[0012] In the above technical solution, the traditional Chinese medicine composition, by weight, is further preferably composed of the following raw materials: 15 parts of Pinellia ternata, 9 parts of Scutellaria baicalensis, 9 parts of dried ginger, 9 parts of Codonopsis pilosula, 9 parts of prepared licorice root, 6 parts of jujube, 3 parts of Coptis chinensis, 15 parts of Dioscorea opposita, 9 parts of stir-fried Atractylodes lancea, 12 parts of Salvia miltiorrhiza, 9 parts of Epimedium brevicornu, and 10 parts of inulin.

[0013] In the above technical solution, the inulin is preferably derived from Jerusalem artichoke or chicory, obtained by hot water extraction, refining, concentration and drying of Jerusalem artichoke or chicory, with a degree of polymerization DP≥3, an inulin-type fructan content of not less than 86%, a moisture content of not more than 5%, and the pollutants and microbial indicators such as lead, arsenic, pathogenic bacteria and so on meet the requirements of the relevant national new resource food or food safety standards.

[0014] The second objective of this invention is to provide a method for preparing a traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome, comprising the following steps:

[0015] Weigh out 13-17 parts of Pinellia ternata, 7-11 parts of Scutellaria baicalensis, 7-11 parts of Zingiber officinale, 7-11 parts of Codonopsis pilosula, 7-11 parts of Glycyrrhiza uralensis (processed), 4-8 parts of Ziziphus jujuba, 1-5 parts of Coptis chinensis, 15-20 parts of Dioscorea opposita, 7-11 parts of Atractylodes lancea (stir-fried with wheat bran), 10-15 parts of Salvia miltiorrhiza, and 7-11 parts of Epimedium brevicornu. Mix them together, add water and decoct 1-3 times, combine the decoctions; filter, concentrate the filtrate under reduced pressure to obtain a clear extract; add ethanol to the clear extract, let it stand, take the supernatant, recover the ethanol and concentrate to obtain an extract; add 8-12 parts of inulin to the extract and stir evenly to obtain the aforementioned traditional Chinese medicine composition.

[0016] In the above technical solution, when adding water and decocting 1 to 3 times, the amount of water added each time is 6 to 12 times the total mass of the raw materials, and each decoction lasts 0.5 to 2 hours.

[0017] In the above technical solution, the decoction is combined and filtered, and the filtrate is concentrated under reduced pressure to a clear extract with a relative density of 1.10 to 1.25 and a temperature of 50 to 65°C.

[0018] In the above technical solution, ethanol is added to the extract to make the ethanol content reach 60% to 80%, and the mixture is allowed to stand for 12 to 36 hours. The supernatant is then taken, the ethanol is recovered and concentrated to obtain an extract with a relative density of 1.25 to 1.35.

[0019] The third objective of this invention is to provide an application of the aforementioned traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome in the preparation of drugs for treating insulin-resistant polycystic ovary syndrome.

[0020] In the above technical solution, the drug for treating insulin-resistant polycystic ovary syndrome is available in any one of the following formulations: decoction, tablets, capsules, lozenges, granules, pills, powders, ointments, elixirs, suspensions, solutions, injections, suppositories, ointments, sprays, drops, drop pills, or patches; preferably, the formulation is an ointment or granules. The method used to prepare the drug formulation of this invention is a conventional method disclosed in the art, and commonly used pharmaceutical excipients in the art can be added.

[0021] In the above technical solution, the drug for treating insulin-resistant polycystic ovary syndrome is used at a dosage of 5-15g (1 sachet) each time, 1-2 times a day, dissolved in a small amount of warm water before taking.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) The traditional Chinese medicine composition of the present invention can effectively alleviate obesity caused by disease model, has a certain protective effect on ovarian function, and has certain potential in regulating reproductive endocrine function.

[0024] (2) The traditional Chinese medicine composition of the present invention can significantly improve the abnormal glucose metabolism of PCOS mice, reduce blood glucose levels and improve insulin sensitivity, thereby alleviating insulin resistance symptoms;

[0025] (3) The traditional Chinese medicine composition of the present invention has the effect of alleviating symptoms and gradually restoring ovarian morphology in PCOS model mice. It shows the best synergistic protective effect in restoring granulosa cell proliferation and arrangement, reducing cystic follicle formation, and promoting ovulation (corpus luteum formation).

[0026] (4) The traditional Chinese medicine composition of the present invention, inulin enhances the pharmacological effects of the traditional Chinese medicine components in improving the ovarian inflammatory microenvironment, regulating endocrine function and promoting follicle development, and effectively reverses the ovarian pathological damage caused by PCOS.

[0027] (5) The traditional Chinese medicine composition of the present invention significantly reduced the levels of inflammatory cytokines such as IL-6, IL-1β and TNF-α in the blood circulation of PCOS mice, indicating that it has anti-inflammatory effects and may further improve the disease process by inhibiting chronic inflammation.

[0028] In summary, the traditional Chinese medicine composition of this invention consists of traditional Chinese medicine decoction pieces and new resource food components. The traditional Chinese medicine decoction pieces are used to exert the therapeutic effects of traditional Chinese medicine syndrome differentiation, while inulin, as a functional dietary fiber component, can adjust the intestinal flora and improve the intestinal environment. It forms a synergy with the traditional Chinese medicine components in terms of formulation structure and mechanism of action, making up for the shortcomings of existing treatment technologies. It has extremely high clinical application value, industrial value and social significance, and this application has a certain degree of innovation. Attached Figure Description

[0029] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0030] Figure 1 To verify the final comparison of the appearance of mice in different groups in Example 1;

[0031] Figure 2 To verify the comparison of body weight changes in different groups of mice in Example 1; where: ● represents the CK group, ■ represents the PCOS group, ▲ represents the PCOS+T group, and ▼ represents the PCOS+T+I group;

[0032] Figure 3 To verify the final comparison of the ovarian appearance of mice in different groups in Example 1;

[0033] Figure 4 To verify the final comparison of ovarian weight in different groups of mice in Example 1;

[0034] Figure 5To verify the comparison of the final body weights of mice in different groups in Example 1;

[0035] Figure 6 To verify the comparison chart of the final ovary / body weight ratio (O / B) of mice in different groups in Example 1;

[0036] Figure 7 To verify the changes in the estrous cycle of mice in different groups in Example 2; where: ● represents the CK group, ■ represents the PCOS group, ▲ represents the PCOS+T group, and ▼ represents the PCOS+T+I group;

[0037] Figure 8 To verify the intraperitoneal glucose tolerance graphs of different groups of mice in Example 3;

[0038] Figure 9 To verify the intraperitoneal insulin tolerance graphs of different groups of mice in Example 3;

[0039] Figure 10 To verify the changes in the morphology of mouse ovarian pathological sections in different groups in Example 4;

[0040] Figure 11 To verify the IL-6 content in the blood of mice in different groups in Example 5;

[0041] Figure 12 To verify the IL-1β content in the blood of mice in different groups in Example 5;

[0042] Figure 13 To verify the TNF-α content in the blood of mice in different groups in Example 5. Detailed Implementation

[0043] Various exemplary embodiments of the present invention are now described in detail. This detailed description should not be considered as a limitation of the invention, but rather as a more detailed description of certain aspects, features, and embodiments of the invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, regarding numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in the invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0044] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0045] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0046] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0047] The present invention will now be described in conjunction with specific embodiments:

[0048] Example 1:

[0049] A traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome includes: 15 g of Pinellia ternata, 9 g of Scutellaria baicalensis, 9 g of dried ginger, 9 g of Codonopsis pilosula, 9 g of prepared licorice root, 6 g of jujube, 3 g of Coptis chinensis, 15 g of Dioscorea opposita, 9 g of stir-fried Atractylodes lancea, 12 g of Salvia miltiorrhiza, 9 g of Epimedium brevicornu, and 10 g of inulin.

[0050] The inulin mentioned above is derived from Jerusalem artichoke, which is obtained by hot water extraction, refining, concentration and drying. It has a degree of polymerization DP≥3, an inulin-type fructan content of not less than 86%, a moisture content of not more than 5%, and the levels of pollutants such as lead, arsenic, pathogenic bacteria and microbial indicators meet the requirements of relevant national standards for new resource foods or food safety standards.

[0051] The traditional Chinese medicine composition is obtained by the following preparation method;

[0052] Accurately weigh the Chinese herbal medicine slices in the formula, mix them, and place them in an extraction tank. First, add 10 times the amount of purified water, soak for 20-40 minutes, heat and decoct for 1.5 hours, filter through a 200-mesh screen, and collect the filtrate. Second, add 8 times the amount of purified water, decoct for 1 hour, and filter. Third, add 6 times the amount of purified water, decoct for 0.5 hours, and filter. Combine the three filtrates and concentrate under reduced pressure at 60-70℃ to a clear extract with a relative density of 1.15-1.20. Add 95% ethanol to the clear extract to achieve an alcohol content of 70%, stir, and let stand for 24 hours. Take the supernatant, recover the ethanol, and continue concentrating to an extract with a relative density of 1.28-1.32. Mix the extract with 10 g of inulin to obtain the aforementioned Chinese herbal medicine composition.

[0053] Add an appropriate amount of maltodextrin to the traditional Chinese medicine composition, mix, prepare a soft material, granulate through a 16-mesh sieve; dry in a fluidized bed at below 60°C, with the moisture content controlled to not exceed 5%; granulate through a 14-mesh sieve, and package to obtain granules for treating insulin-resistant polycystic ovary syndrome, with each bag containing 5-15 g.

[0054] To verify the efficacy of the traditional Chinese medicine composition of this invention, a high-fat diet combined with DHEA-induced polycystic ovary syndrome (PCOS) mouse model was established to investigate the therapeutic effect of the traditional Chinese medicine + novel resource food composition on insulin-resistant PCOS. The composition was tested for its basic characteristics in terms of weight control and ovarian weight, further verifying its protective effect on ovarian function. In addition, its potential to improve estrous cycle disorders in mice was examined. Simultaneously, glucose tolerance and insulin tolerance tests were conducted to determine whether it could alleviate insulin resistance symptoms. Most importantly, ovarian pathological sections were observed to confirm the pathological changes in the PCOS model and different groups. Finally, the detection of inflammatory factors confirmed that the composition reduced the levels of inflammatory cytokines such as IL-6, IL-1β, and TNF-α in the circulating blood of mice.

[0055] The following verification examples will be used to verify the efficacy of the traditional Chinese medicine composition of the present invention in improving insulin-resistant polycystic ovary syndrome in mice:

[0056] Verification Example 1: Effect of Traditional Chinese Medicine Composition on Mouse Body Weight

[0057] I. Experimental Materials and Grouping:

[0058] 1.1 Experimental animals:

[0059] Twenty-four 3-week-old female C57BL / 6 mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed in a laboratory with a constant temperature of 20-25℃ and humidity of 40-60%, with alternating 12-hour light and 12-hour dark cycles daily. The mice had free access to food and water during the experiment. This study was approved by the Ethics Committee of Tianjin Jinke Bona Biotechnology Co., Ltd., approval number: GENINK-20250039.

[0060] 1.2 Animal grouping and model construction:

[0061] Twenty-four 3-week-old female C57BL / 6 mice were acclimatized for one week under natural light and free access to water and food. Then, 18 mice were randomly selected and given daily subcutaneous injections of DHEA (60 mg / kg, dissolved in sterile sesame oil, 0.1 ml / mouse) via the back of the neck, in combination with a high-sugar, high-fat diet. The remaining 6 mice were given the same amount of sterile sesame oil daily subcutaneously via the back of the neck and fed a normal diet.

[0062] 1.3 The specific details of group processing and design are as follows:

[0063] Control group (CK group) (injected with an equal volume of sterile sesame oil): 6 animals, given a normal diet;

[0064] PCOS group (daily DHEA injection): 6 animals, given a high-fat diet (60% high-fat).

[0065] PCOS+T group (daily DHEA injection): 6 animals were given a high-fat diet (60% high-fat) and then simultaneously administered a pure traditional Chinese medicine composition by gavage daily (dosage: 5.2 g / kg, freshly prepared or refrigerated at 4°C, administered at a fixed time each day); the preparation method of the pure traditional Chinese medicine composition was basically the same as that of the traditional Chinese medicine composition in Example 1, except that inulin was not added.

[0066] The PCOS+T+I group (daily DHEA injection): 6 animals were given a high-fat diet (60% high-fat) and a combination of traditional Chinese medicine and inulin administered by gavage daily (dosage: 5.2 g / kg, freshly prepared or refrigerated at 4°C, administered at a fixed time each day). The preparation method of the traditional Chinese medicine + inulin combination was the same as that of the traditional Chinese medicine combination in Example 1.

[0067] II. Experimental Methods:

[0068] Monitoring of mouse appearance and weight changes: Mouse weight was measured using an electronic balance, and the trend of weight change was recorded and analyzed. Mice were first monitored for weight one week after acclimatization, and then monitored every 3 days at the same time until the end of the experiment. A line graph of mouse weight was plotted to observe weight changes, and the appearance and condition of the mice were observed and recorded. After the experiment, mice were euthanized by cervical dislocation, and both ovaries were quickly removed and weighed; the ratio of ovary weight to body weight was calculated.

[0069] All data were statistically analyzed using SPSS 25 software. Results are expressed as mean ± standard deviation. The least significant difference (LSD) method was used to calculate significance. One-way ANOVA was used to analyze differences between groups. P < 0.05 was considered statistically significant. Plots were generated using OriginPro 2019b 64-Bit.

[0070] III. Experimental Results:

[0071] like Figure 1 , Figure 2 and Figure 5 As shown, at the beginning of the experiment, there was no significant difference in the average body weight of the mice in each group, indicating a consistent initial baseline. As the experiment progressed, the body weight of the PCOS group mice significantly increased compared to the CK group, a phenomenon consistent with the characteristics of a high-fat diet combined with DHEA-induced polycystic ovary syndrome (PCOS) model. Specifically, at week 3, the average body weight of the PCOS group mice increased by approximately 25% compared to the CK group, a statistically significant difference (P<0.05). The body weight of the mice in the PCOS+T+I group was significantly lower than that of the PCOS group and approached the level of the CK group, suggesting that the traditional Chinese medicine composition can effectively inhibit the obesity phenotype induced by a high-fat diet and drug-induced modeling. Furthermore, the PCOS+T group also showed some weight-inhibiting effect, but the effect was not as significant as that of the PCOS+T+I group. These results indicate that the traditional Chinese medicine composition may alleviate obesity symptoms in PCOS model mice by regulating metabolic function or improving insulin resistance.

[0072] After the experiment, the weight of the mouse ovaries was measured. Figure 3 and Figure 4 The results showed that the ovarian weight of mice in the PCOS group was significantly higher than that in the CK group (P<0.01), which is consistent with the ovarian morphological changes commonly seen in patients with polycystic ovary syndrome. Further analysis revealed that the ovarian weight of mice in the PCOS group increased by approximately 40% compared to the CK group, and the ovarian-to-body weight ratio was also significantly higher, indicating that the increase in ovarian weight was not solely due to weight gain. In contrast, the ovarian weight of mice in the PCOS+T+I group was significantly lower than that in the PCOS group, and there was no significant difference compared to the CK group (P>0.05). The ovarian-to-body weight ratio was calculated... Figure 6This allows for a more direct observation that the ovarian weight in the treatment groups (PCOS+T+I group and PCOS+T group) essentially returned to normal levels, further validating the ameliorative effect of the traditional Chinese medicine composition on ovarian morphological abnormalities. These results suggest that the traditional Chinese medicine composition may alleviate the increase in ovarian weight in PCOS model mice by regulating hormone levels or inhibiting local ovarian inflammation.

[0073] Verification Example 2: Effects of Traditional Chinese Medicine Composition on Estrogenic Cycle Changes in Mice

[0074] I. Experimental Materials and Grouping:

[0075] Same as verification example one.

[0076] II. Experimental Methods:

[0077] Changes in estrous cycle in mice of different groups: Seven days after the start of mouse intervention modeling, the vagina of mice was irrigated with physiological saline from 10:00 to 12:00 every day to collect vaginal cells for smear preparation and staining with methylene blue. The slides were then placed under a microscope for observation. This was carried out for 14 consecutive days. The results were plotted as a line graph of the estrous cycle to observe whether the estrous cycle was disordered.

[0078] Data processing is the same as in Implementation Example 1.

[0079] III. Experimental Results:

[0080] This experiment used DHEA to establish a PCOS mouse model. The success of the PCOS mouse model was determined by observing changes in the estrous cycle of the mice. Furthermore, the therapeutic effect of the traditional Chinese medicine composition described in this invention on the PCOS model mice was observed. The estrous cycle of normal mice is generally 4-5 days, with each estrous phase occurring periodically, and all cell types showing normal developmental morphology. The results showed ( Figure 7 In the control group (CK group), mice exhibited normal estrous cycles, each lasting 4-5 days, with a relatively consistent proportion of mice in different estrous cycle stages. Mice in the PCOS group showed estrous cycle disorder, with most mice in the post-estrus or anteroestrus phase, and the cycles were prolonged and irregular. After treatment with the traditional Chinese medicine component in DHEA-induced PCOS mice, daily observation of vaginal smears in the PCOS+T+I and PCOS+T groups showed that the estrous cycle essentially returned to normal. These results suggest that the traditional Chinese medicine component developed in this invention has a mitigating effect on the estrous cycle in PCOS mice. These results indicate that the traditional Chinese medicine composition may improve the reproductive endocrine disorder in PCOS model mice by regulating the function of the hypothalamus-pituitary-ovarian axis.

[0081] Verification Example 3: Effects of Traditional Chinese Medicine Composition on Changes in Peritoneal Glucose Tolerance and Insulin Tolerance in Mice

[0082] I. Experimental Materials and Grouping:

[0083] Same as verification example one.

[0084] II. Experimental Methods:

[0085] Changes in intraperitoneal glucose tolerance and insulin tolerance in mice: If the estrous cycle of mice is indeed disrupted, indicating that the current modeling effect meets our expectations, then the Intraperitoneal Glucose Tolerance Test (IPGTT) and Insulin Tolerance Test (ITT) will be conducted; otherwise, the modeling time will be appropriately increased. At 17:00, mice were fasted but allowed water. The following morning at 8:00, 6 mice in each group were intraperitoneally injected with a 20% glucose solution based on body weight (2g / kg glucose). Blood glucose levels were measured and recorded at 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min after injection. On the third day at 9:00 AM, mice were fasted but allowed water. At 14:00 PM, 6 mice in each group were intraperitoneally injected with a 0.75 U / kg insulin solution based on body weight. Blood samples were collected, tested, and recorded at the same 6 time points.

[0086] Data processing is the same as in Implementation Example 1.

[0087] III. Experimental Results:

[0088] The results of the intraperitoneal glucose tolerance test showed that... Figure 8 and Figure 9 As shown, after glucose injection, the blood glucose level in the PCOS group mice rapidly increased and was significantly higher than that in the other three groups at all time points (P<0.05), especially at 15 and 60 minutes, indicating that the mice in this group had significant glucose tolerance abnormalities. In contrast, the blood glucose levels in the traditional Chinese medicine (TCM) intervention groups (PCOS+T+I group and PCOS+T group) were closer to those in the control (CK) group at all time points, indicating that the TCM composition could significantly improve the glucose metabolism capacity of PCOS model mice. The results of the insulin tolerance test further supported this conclusion: the decrease in blood glucose after insulin administration in the PCOS group mice was significantly lower than that in the other three groups, while the TCM intervention groups showed a similar trend in blood glucose dynamics as the CK group. These results suggest that the TCM composition may improve the insulin resistance status of PCOS model mice by enhancing insulin sensitivity or promoting glucose uptake and utilization.

[0089] Verification Example 4: Effects of Traditional Chinese Medicine Composition on Pathological Changes in Mouse Ovarian Tissue

[0090] I. Experimental Materials and Grouping:

[0091] Same as verification example one.

[0092] II. Experimental Methods:

[0093] Pathological specimen collection of mouse ovarian tissue: After the experiment, mice were euthanized, and the entire experimental process complied with animal ethics guidelines. Mice were fasted for 8 hours after the experiment and anesthetized with 3% sodium pentobarbital via intraperitoneal injection. After anesthesia took effect, the abdominal wall of the mice was cut open with scissors. The cecum was dissected, and feces were collected into sterile cryovials, sealed, and stored at -80°C. Both ovaries of the mice were harvested, photographed, and weighed. The right ovary was fixed with 4% paraformaldehyde, and the left ovary was photographed and placed in cryovials for storage at -80°C.

[0094] II. Experimental Results:

[0095] To investigate the effects of the traditional Chinese medicine composition developed in this invention on the morphology of mouse ovaries, paraffin sections of ovarian tissue were prepared, and the morphology of the ovarian tissue was observed under an optical microscope. The results showed that ( Figure 10 CK group: The overall ovarian structure is basically normal, with primary follicles, secondary follicles (as shown by the yellow arrows in the figure), and mature follicles (as shown by the black arrows in the figure) visible within the tissue. PCOS group: The overall ovarian tissue structure is abnormal, with atretic follicles (as shown by the yellow arrows in the figure) and corpus luteum (as shown by the black arrows in the figure) visible within the tissue. PCOS+T group: The overall ovarian tissue structure is abnormal, with a large number of atretic follicles (as shown by the yellow arrows in the figure) and corpus luteum (as shown by the black arrows in the figure) visible within the tissue. PCOS+T+I group: The overall ovarian structure is slightly abnormal, with primary follicles, secondary follicles (as shown by the yellow arrows in the figure), mature follicles (as shown by the black arrows in the figure), and atretic follicles (as shown by the blue arrows in the figure) visible within the tissue.

[0096] These four H&E-stained sections of mouse ovarian tissue (scale bar 100 μm each) morphologically and directly reflect the effectiveness of the polycystic ovary syndrome (PCOS) model and the therapeutic effects of different intervention regimens. CK group ( Figure 8 (Left upper side) Pathological features: The ovarian tissue structure is intact, and normal follicles at different developmental stages (including primary, secondary, and mature follicles) are visible. Cell morphology: The granulosa cell layer of each follicle is neatly arranged and thick, with multiple cell layers; the oocytes are clearly morphological and show no degeneration, and the cumulus oophorus structure is normal. Ovulation indicators: Numerous corpora lutea (light purple irregular cell clusters) are visible in the field of view, indicating normal ovulation function. The ovarian matrix is ​​loose, and no obvious hyperplasia is observed. PCOS model group ( Figure 8(Upper right side) Pathological features: Typical PCOS ovarian morphological changes were observed. Numerous cystic dilated follicles were present in the field of view, with significantly enlarged follicular cavities. Cell morphology: The most prominent feature was the thinning or even complete shedding of the granulosa cell layer, leaving only a very thin layer or even a single layer of granulosa cells; many cystic follicles lacked oocytes, or the oocytes underwent atresia and degeneration. Ovulation indicators: The number of corpora lutea was significantly reduced or even absent, suggesting ovulation disorders or infrequent ovulation. The ovarian stromal cells appeared slightly dense, suggesting possible stromal hyperplasia. The model was successfully established. PCOS+T group ( Figure 8 (Lower left side) Pathological features: Compared with the model group, the number and volume of cystic follicles in the ovary were reduced, but some cystic dilated vacuoles were still visible. Cell morphology: The granulosa cell layer was significantly thickened compared with the model group, the cells were arranged relatively regularly, and some follicles contained oocytes. Efficacy evaluation: The PCOS+T group alone could partially improve follicular development arrest and granulosa cell reduction caused by PCOS, and had a certain repair effect on ovarian function, but it has not yet fully recovered to the normal physiological state (residual atresia or cystic structures can still be seen). PCOS+T+I group ( Figure 8 (Lower right side) Pathological features: The pathological morphology of the ovarian tissue was significantly improved and optimized, with a substantial reduction in the number of cystic dilated follicles and the disappearance of most vacuolar structures. Cell morphology: The number of normally developing follicles at each stage increased significantly. Most notably, the granulosa cell layer recovered to a multi-layered and densely arranged healthy state, tightly enveloping the oocyte; the cumulus oophorus cell structure was normal, and follicle development was good. Ovulation indicators: Normal corpus luteum formation was visible in the observation field, with fewer atretic follicles, and no hyperplasia was observed in the ovarian stroma. The overall morphology was closest to the blank control group.

[0097] Based on the above pathological section observations, the following conclusions can be drawn: The PCOS model was successfully established. The PCOS group exhibited typical characteristics of cystic follicular dilation, granulosa cell apoptosis / thinning, and ovulation disorders. The traditional Chinese medicine (TCM) composition group showed a trend of progressively decreasing lesion severity and gradual recovery of ovarian morphology. It demonstrated the best synergistic protective effect in restoring granulosa cell proliferation and arrangement, reducing cystic follicle formation, and promoting ovulation (corpus luteum formation). The addition of inulin may have enhanced the pharmacological effects of the TCM composition in improving the ovarian inflammatory microenvironment, regulating endocrine function, and promoting follicle development through pathways such as regulating the intestinal microecology, effectively reversing the ovarian pathological damage caused by PCOS.

[0098] Verification Example 5: Effects of Traditional Chinese Medicine Composition on Changes in Blood Inflammatory Factors in Mice

[0099] I. Experimental Materials and Grouping:

[0100] Same as verification example one.

[0101] II. Experimental Methods:

[0102] Measurement of Inflammatory Factors in Mouse Blood: The levels of inflammatory factors IL-6, IL-1β, and TNF-α in mouse blood circulation were detected using enzyme-linked immunosorbent assay (ELISA). After the experiment, blood was collected from mice using the orbital venous plexus method. After standing at room temperature for 2 hours, the serum was separated by centrifugation at 3000 rpm for 15 minutes. The concentrations of each inflammatory factor were measured according to the ELISA kit instructions. The ELISA kit, purchased from R&D Systems, was used to detect serum levels of sex hormones and the expression levels of inflammatory factors IL-6, IL-1β, and TNF-α.

[0103] Data processing is the same as in Implementation Example 1.

[0104] III. Experimental Results:

[0105] Effects of different groups of inflammatory factors in mice: Chronic inflammation plays an important role in the development of polycystic ovary syndrome. This study assessed the inflammatory status of mice in different groups by detecting the levels of inflammatory factors IL-6, IL-1β, and TNF-α in the blood circulation. The results showed ( Figure 11 , Figure 12 and Figure 13 In the PCOS group, the levels of IL-6, IL-1β, and TNF-α were significantly higher than those in the CK group (P<0.01), consistent with the reported elevated levels of chronic inflammation in PCOS patients. The levels of inflammatory factors in the traditional Chinese medicine (TCM) intervention groups (PCOS+T and PCOS+T+I groups) were significantly reduced, especially in the PCOS+T+I group, where the inflammatory factor levels were close to those in the CK group. These results suggest that the TCM composition may alleviate the chronic inflammatory state in PCOS model mice by inhibiting the release of inflammatory factors or regulating related signaling pathways, providing important molecular mechanistic support for its therapeutic effect.

[0106] As described above, this invention establishes a high-fat diet combined with DHEA-induced polycystic ovary syndrome (PCOS)-like mouse model with abnormal glucose and lipid metabolism, and explores the therapeutic effect of a traditional Chinese medicine (TCM) + novel resource food composition on insulin-resistant PCOS. Experimental results show that the composition exhibits significant therapeutic effects on several key indicators. First, regarding weight control, the PCOS group mice had significantly higher body weight than the control group (CK group), while the TCM + novel resource food composition intervention group (PCOS+T+I group) showed significantly inhibited weight gain, suggesting that the composition can effectively alleviate obesity induced by the disease model. Second, regarding ovarian weight, the PCOS group mice had significantly higher ovarian weight than the CK group, while the ovarian weight significantly decreased after composition intervention, approaching that of the normal group, further verifying the protective effect of the composition on ovarian function. Furthermore, the composition significantly improved the estrous cycle disorder in mice, restoring the PCOS group mice to near-normal cyclical changes, providing strong evidence for the composition's potential in regulating reproductive endocrine function. Simultaneously, glucose tolerance and insulin tolerance tests showed that the composition significantly improved glucose metabolism abnormalities in PCOS mice, reduced blood glucose levels, and increased insulin sensitivity, thereby alleviating insulin resistance symptoms. Combined with the above pathological section observations, the successful establishment of the PCOS model was confirmed. The PCOS group exhibited typical characteristics of cystic follicle dilation, granulosa cell apoptosis / thinning, and ovulation disorders. The composition group showed a trend of progressively decreasing lesion severity and gradual recovery of ovarian morphology. It demonstrated the best synergistic protective effect in restoring granulosa cell proliferation and arrangement, reducing cystic follicle formation, and promoting ovulation (corpus luteum formation). The addition of inulin may have enhanced the pharmacological effects of the simple Chinese medicine group (PCOS+T group) in improving the ovarian inflammatory microenvironment, regulating endocrine function, and promoting follicle development through pathways such as regulating the intestinal microecology, effectively reversing the ovarian pathological damage caused by PCOS. Finally, the inflammatory factor detection results showed that the composition significantly reduced the levels of inflammatory cytokines such as IL-6, IL-1β, and TNF-α in the blood circulation of PCOS mice, indicating its anti-inflammatory effect, which may further improve the disease process by inhibiting chronic inflammation.

[0107] In conclusion, this study fully demonstrates the effectiveness of the combination of traditional Chinese medicine and new resource food in improving symptoms associated with insulin-resistant polycystic ovary syndrome.

[0108] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome, characterized in that, The active pharmaceutical ingredient, by weight, includes the following parts: Components of the prepared Chinese herbal medicine: 13-17 parts of Pinellia ternata, 7-11 parts of Scutellaria baicalensis, 7-11 parts of dried ginger, 7-11 parts of Codonopsis pilosula, 7-11 parts of prepared licorice root, 4-8 parts of jujube, 1-5 parts of Coptis chinensis, 15-20 parts of Dioscorea opposita, 7-11 parts of stir-fried Atractylodes lancea, 10-15 parts of Salvia miltiorrhiza, and 7-11 parts of Epimedium brevicornu; New resource food component: 8-12 parts inulin.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The raw materials include the following parts by weight: 14-16 parts of Pinellia ternata, 8-10 parts of Scutellaria baicalensis, 8-10 parts of dried ginger, 8-10 parts of Codonopsis pilosula, 8-10 parts of prepared licorice root, 5-7 parts of jujube, 2-4 parts of Coptis chinensis, 15-18 parts of Dioscorea opposita, 8-10 parts of stir-fried Atractylodes lancea, 11-14 parts of Salvia miltiorrhiza, 8-10 parts of Epimedium brevicornu, and 9-11 parts of inulin.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The raw materials include the following parts by weight: 15 parts of Pinellia ternata, 9 parts of Scutellaria baicalensis, 9 parts of dried ginger, 9 parts of Codonopsis pilosula, 9 parts of prepared licorice root, 6 parts of jujube, 3 parts of Coptis chinensis, 15 parts of Dioscorea opposita, 9 parts of stir-fried Atractylodes lancea, 12 parts of Salvia miltiorrhiza, 9 parts of Epimedium brevicornu, and 10 parts of inulin.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, The inulin mentioned above is derived from Jerusalem artichoke or chicory, obtained by hot water extraction, refining, concentration, and drying of Jerusalem artichoke or chicory. It has a degree of polymerization DP≥3, an inulin-type fructan content of not less than 86%, a moisture content of not more than 5%, and the levels of pollutants such as lead, arsenic, pathogenic bacteria, and microbial indicators meet the requirements of relevant national standards for new resource foods or food safety standards.

5. A method for preparing a traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome according to any one of claims 1 to 4, characterized in that, Includes the following steps: Weigh out 13-17 parts of Pinellia ternata, 7-11 parts of Scutellaria baicalensis, 7-11 parts of Zingiber officinale, 7-11 parts of Codonopsis pilosula, 7-11 parts of Glycyrrhiza uralensis (processed), 4-8 parts of Ziziphus jujuba, 1-5 parts of Coptis chinensis, 15-20 parts of Dioscorea opposita, 7-11 parts of Atractylodes lancea (stir-fried with wheat bran), 10-15 parts of Salvia miltiorrhiza, and 7-11 parts of Epimedium brevicornu. Mix them together, add water and decoct 1-3 times, combine the decoctions; filter, concentrate the filtrate under reduced pressure to obtain a clear extract; add ethanol to the clear extract, let it stand, take the supernatant, recover the ethanol and concentrate to obtain an extract; add 8-12 parts of inulin to the extract and stir evenly to obtain the aforementioned traditional Chinese medicine composition.

6. The preparation method according to claim 5, characterized in that, When decocting with water 1 to 3 times, the amount of water added each time should be 6 to 12 times the total mass of the raw materials, and each decoction should last 0.5 to 2 hours.

7. The preparation method according to claim 5, characterized in that, After combining the decoctions, filter them and concentrate the filtrate under reduced pressure to obtain a clear extract with a relative density of 1.10–1.25 and a temperature of 50–65°C. Add ethanol to the clear extract to make the ethanol content reach 60%–80%, let it stand for 12–36 hours, take the supernatant, recover the ethanol and concentrate it to obtain an extract with a relative density of 1.25–1.

35.

8. The use of the traditional Chinese medicine composition for treating insulin-resistant polycystic ovary syndrome according to any one of claims 1 to 4 in the preparation of a drug for treating insulin-resistant polycystic ovary syndrome.

9. The application according to claim 8, characterized in that, The drug for treating insulin-resistant polycystic ovary syndrome is available in any one of the following formulations: decoction, tablets, capsules, lozenges, granules, pills, powders, ointments, elixirs, suspensions, solutions, injections, suppositories, ointments, sprays, drops, drop pills, or patches; preferably, the formulation is an ointment or granules.

10. The application according to claim 8, characterized in that, The medication for treating insulin-resistant polycystic ovary syndrome is used at a dosage of 5-15g each time, 1-2 times daily, dissolved in a small amount of warm water before taking.

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