Application of jingye grass extract in prevention and treatment of polycystic ovary syndrome

Abstract

The present application relates to the application of the extract of the plant of Aneilema aescynum in preventing and treating polycystic ovary syndrome. The present application finds that the extract of the plant of Aneilema aescynum can effectively inhibit systemic inflammation of polycystic ovary syndrome (PCOS) mice, antagonize oxidative stress, and ultimately positively regulate sex hormone balance (increase E2 level, and reduce AMH and testosterone level), effectively improve ovulation dysfunction. The extract of the plant of Aneilema aescynum can improve the pathophysiological basis of PCOS through anti-inflammatory, antioxidant and metabolic regulation, so as to restore the homeostasis of reproductive endocrine, and effectively treat PCOS.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of *Chicken Eye Grass* extract in the prevention and treatment of polycystic ovary syndrome. Background Technology

[0002] The ovary is a female reproductive organ, belonging to the female gonads. Its main function is to produce and release egg cells, and it also secretes sex hormones. Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. Its pathogenesis and mechanisms are not yet fully understood, but current research suggests a possible link between genetics and environmental metabolism. Therefore, there are currently no effective treatment options available domestically or internationally, and the search for specific and effective targeted therapies remains.

[0003] Because the ovaries in a woman's body are responsible for secreting estrogen, polycystic ovary syndrome (PCOS) can directly affect estrogen levels and indirectly influence the synthesis and release of other hormones, leading to diseases such as type 2 diabetes, obesity, and atherosclerosis. Furthermore, excessive androgen secretion can also cause infertility.

[0004] Although various medications are currently used to treat polycystic ovary syndrome (PCOS), its pathogenesis remains unclear, and estrogen has multiple effects on various organs. Therefore, existing medications are primarily used clinically for symptomatic treatment. Based on clinical symptoms, they can be categorized into medications that regulate the menstrual cycle, lower blood androgen levels, improve insulin resistance, and induce ovulation.

[0005] Currently, there is a lack of drugs that can comprehensively improve the pathophysiological basis of PCOS and restore reproductive endocrine homeostasis. Therefore, there is still a need in this field to develop drugs that can provide systemic treatment for PCOS. Summary of the Invention

[0006] Therefore, the purpose of this invention is to provide the application of *Centipeda minima* extract in the prevention and treatment of polycystic ovary syndrome (PCOS). *Centipeda minima* extract can comprehensively improve the pathophysiological basis of PCOS, restore reproductive endocrine homeostasis, and has a good therapeutic effect on PCOS.

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

[0008] In a first aspect, the invention provides the use of an extract of *Kummerowia striata* (Thunb.) Schindl in the preparation of a medicament for the prevention and / or treatment of polycystic ovary syndrome.

[0009] In a second aspect, the invention provides the use of *Centipeda minima* extract in combination with hormonal drugs, insulin sensitizers, and / or ovulation-inducing drugs in the preparation of a medicament for the prevention and / or treatment of polycystic ovary syndrome.

[0010] In some embodiments, the cornflower extract includes cornflower water extract, cornflower alcohol extract, and cornflower water-alcohol extract.

[0011] In some embodiments, the cornflower extract is a water extract of cornflower.

[0012] In some embodiments, the water extract of *Chicken Eye Grass* is prepared by the following method: *Chicken Eye Grass* is taken, crushed to obtain coarse powder, then water is added and refluxed for extraction 1 to 3 times, the filtrates are combined, concentrated under reduced pressure, and dried to obtain the extract.

[0013] In some embodiments, the mass ratio of the coarse powder to water is 1:(8~12), preferably 1:(8~10).

[0014] In some embodiments, the reflux extraction temperature is 110℃~130℃ and the time is 1h~2h; preferably, the reflux extraction temperature is 115℃~125℃ and the time is 1h~1.5h.

[0015] In some implementations, the application includes regulating hormone levels associated with polycystic ovary syndrome (PCOS).

[0016] In some implementations, the application includes increasing estradiol levels, decreasing anti-Müllerian hormone levels, and / or decreasing testosterone levels.

[0017] In some implementations, the application includes improving polycystic ovary syndrome-related oxidative stress levels and metabolic abnormalities.

[0018] In some implementations, the application includes reducing malondialdehyde levels, increasing glutathione levels, and / or reducing body weight.

[0019] In some implementations, the application includes suppressing chronic inflammation associated with polycystic ovary syndrome.

[0020] In some embodiments, the application includes inhibiting the expression levels of pro-inflammatory factors TNF-α, IL-6, and / or IL-1β.

[0021] In some implementations, the application includes improving the clinical symptoms of polycystic ovary syndrome.

[0022] In some implementations, the application includes improving oligomenorrhea, amenorrhea, irregular bleeding, hirsutism, acne, ovulation disorders, and / or polycystic ovarian changes.

[0023] In some embodiments, the dosage form of the drug includes oral and injectable formulations.

[0024] In some embodiments, the oral dosage form includes tablets, capsules, granules, solutions, powders, emulsions, suspensions, decoctions, pills, and ointments.

[0025] A third aspect of the present invention provides a method for preventing and / or treating polycystic ovary syndrome, comprising the steps of: administering a cornflower extract to a person or animal with polycystic ovary syndrome.

[0026] Compared with the prior art, the present invention has the following beneficial effects.

[0027] This invention, through research, has found that *Centella asiatica* extract can effectively inhibit systemic inflammation and antagonize oxidative stress in mice with polycystic ovary syndrome (PCOS), and ultimately positively regulate sex hormone balance (increasing E2 levels and decreasing AMH and testosterone levels), effectively improving ovulation failure. This indicates that *Centella asiatica* extract can comprehensively improve the pathophysiological basis of PCOS through anti-inflammatory, antioxidant, and metabolic regulation, thereby restoring reproductive endocrine homeostasis and effectively treating PCOS.

[0028] Chicken eye grass extract has the characteristics of long-term and mild effects, and is expected to become a new generation of traditional Chinese medicine treatment for PCOS. Attached Figure Description

[0029] Figure 1 It is called chicken-eye grass.

[0030] Figure 2 The changes in serum E2 in female mice from different groups.

[0031] Figure 3 The changes in serum AMH in female mice from different groups.

[0032] Figure 4 The changes in serum testosterone in female mice from different groups.

[0033] Figure 5 The changes in MDA content in liver tissue homogenates from different groups of female mice were shown.

[0034] Figure 6 Changes in GSH in liver tissue homogenates from different groups of female mice.

[0035] Figure 7 Changes in IL-6 in liver tissue homogenates from different groups of female mice.

[0036] Figure 8 Changes in TNF-alpha in liver tissue homogenates from different groups of female mice.

[0037] Figure 9 Changes in IL-1beta in liver tissue homogenates from different groups of female mice.

[0038] Figure 10 The appearance and pathological sections of the ovaries of female mice from different groups. Detailed Implementation

[0039] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0040] In this invention, the terms "interleukin-1β (IL-1beta)" and "interleukin-6 (IL-6)" are inflammatory factors in the body that can activate downstream signaling pathways and cause inflammatory responses in the body.

[0041] In this invention, the term "anti-Müllerian hormone (AMH)" refers to a hormone in the body that can reflect changes in ovarian function in a woman.

[0042] In this invention, the term "estradiol (E2)" refers to the most important and potent estrogen, and is an important indicator of ovarian function.

[0043] In this invention, the terms "reduced glutathione (GSH) and malondialdehyde (MDA)" are important indicators of oxidative stress.

[0044] In this invention, the term "(α-tumor necrosis factor, TNF-α)" refers to a pro-inflammatory factor secreted by immune cells, and TNF-α levels can be abnormally elevated in chronic inflammatory diseases.

[0045] The term "prevention and treatment" refers to the prevention and / or treatment of diseases or symptoms described in this invention, which can be achieved in subjects to treat, prevent, reduce and / or alleviate the disease or symptom described in this invention.

[0046] The term "disease and / or symptom" refers to a physical condition of the subject that is related to the disease and / or symptom described in this invention.

[0047] The following description is based on specific implementation methods.

[0048] Experimental materials Animals: SPF-grade female KM mice, weighing 18-20g, 3 weeks old, provided by the Animal Experiment Center of Guilin Medical University.

[0049] Chicken eye grass refers to the whole plant of chicken eye grass, including the stems and leaves, which is the dried product obtained after fresh harvesting.

[0050] Chicken-eye grass, scientific name: *Kummerowia striata* (Thunb.) Schindl., also known as three-leafed clover, is a plant belonging to the genus *Kummerowia* in the legume family, such as... Figure 1 As shown.

[0051] Example 1: Preparation of Chicken Eye Grass Extract This embodiment provides a *Chicken Eye Grass* extract, which is prepared by the following method: (1) Preliminary crushing: Take the whole plant of chicken eye grass and crush it into coarse powder.

[0052] (2) Pure water heating and reflux extraction: Take the dried crude powder of Chinese medicine, add 8 times the mass of pure water, reflux extract at 120℃ for 1 hour, filter, add 8 times the mass of pure water again, reflux at 120℃ for 1 hour, and combine the two filtrates after filtration.

[0053] (3) Filtrate concentration and drying: The above filtrate was concentrated under reduced pressure at 60°C and then spray-dried to obtain the extract of *Chicken Eye Grass*.

[0054] 1g of the *Chicken Eye Grass* extract is equivalent to 50g of the original *Chicken Eye Grass* herb.

[0055] Experiment Example 2: Investigation into the effect of *Chicken Eye Grass* extract on letrozole-induced polycystic ovary syndrome in mice Letrozole is a new generation aromatase inhibitor with the chemical name 4,4'-(1H-1,2,4-triazol-1-yl-methylene)-diphenylnitrile. Letrozole is commonly used clinically to treat breast cancer, especially postmenopausal breast cancer.

[0056] Studies have shown that letrozole can inhibit the activity of aromatase, thereby inhibiting the conversion of testosterone and androstenedione into estradiol and estrone, respectively. This increases androgen levels in the body, leading to steroid hormone metabolism disorders and thus establishing an animal model of polycystic ovary syndrome.

[0057] In this embodiment, letrozole was administered within 1-2 hours after modeling to simulate a clinical treatment regimen, thereby determining the therapeutic effect of cornflower extract (hereinafter referred to as KS) on polycystic ovary syndrome.

[0058] I. Experimental Methods 1. Grouping, Modeling, and Drug Administration (1) Grouping Twenty-five 3-week-old female KM mice were randomly divided into five groups: control group, model group, low-dose group (1 g / kg KS), medium-dose group (2 g / kg KS), and high-dose group (3 g / kg KS), with five mice in each group.

[0059] (2) Establishment and administration of letrozole-induced polycystic ovary syndrome model The control group received only the corresponding dose of normal saline by gavage twice a day for 21 consecutive days, without any letrozole or other drug treatment.

[0060] The model group was given a corresponding dose of physiological saline by gavage once a day for 21 consecutive days, based on their body weight.

[0061] Starting from day one, mice in the model group, low-dose group, medium-dose group, and high-dose group were administered letrozole by gavage at a dose of 1 mg / kg for 21 consecutive days, according to their respective body weights, once daily for 21 days until samples were collected. Two hours after administration of letrozole, the high-dose group, medium-dose group, and low-dose group (the three groups being the administration groups) were administered the *Centella asiatica* extract prepared in Example 1, once daily for 21 consecutive days. The specific gavage doses for the *Centella asiatica* extract groups were as follows: high-dose group: 3 g / kg of *Centella asiatica* extract, equivalent to 150 g / kg of concentrated and purified raw material; medium-dose group: 2 g / kg of *Centella asiatica* extract, equivalent to 100 g / kg of concentrated and purified raw material; low-dose group: 1 g / kg of *Centella asiatica* extract, equivalent to 50 g / kg of concentrated and purified raw material.

[0062] Gavage procedure: Mice were weighed daily before gavage to determine the volume of fluid to be administered. Saline, letrozole, and *Centella asiatica* extract were administered at a 1:10 ratio, i.e., 1g of mouse was given 10ul of fluid. The treatment groups received letrozole first, followed by *Centella asiatica* extract; the model group received letrozole first, followed by saline; and the control group received saline twice. There was a 2-hour interval between the two gavage administrations for all groups.

[0063] Starting from day one, mice in the model group, low-dose group, medium-dose group, and high-dose group were fed a diet consisting of 60% high-fat food daily, while the control group was fed a normal diet. Both groups had normal access to water until sample collection.

[0064] 2. Sample collection Twenty-one days after the start of the experiment, blood was collected from the eyeballs, allowed to stand at room temperature for about 1-2 hours, centrifuged at 12,000 rpm, and the supernatant, i.e., serum, was collected and stored at -80°C. Mice were then euthanized by dislocation, and the liver was removed. The liver tissue was chopped, flash-frozen in liquid nitrogen, and then stored at -80°C. At the same time, the ovaries were removed and fixed with 4% paraformaldehyde for subsequent pathological staining.

[0065] 3. Processing of pathological sections Ovarian tissues from different mice were fixed in 4% paraformaldehyde, dehydrated, cleared, embedded, and stored at room temperature. When needed, sections (4 μm) were prepared. Before staining, the sections were baked in an oven at 70°C for 2 hours, followed by dewaxing with xylene, hematoxylin-eosin (HE) staining, dehydration with gradients of different concentrations of ethanol, clearing with xylene, mounting with neutral resin, and observation under an optical microscope.

[0066] 4. Detection of oxidative stress and inflammatory factor markers Accurately weigh liver tissue, add 9 times the volume of physiological saline, and prepare a 10% liver tissue homogenate under ice bath conditions. Centrifuge at 12,000 rpm / min for 15 min at 4°C and collect the supernatant. Determine the levels of GSH, MDA, IL-6, IL-1beta, and TNF-alpha strictly according to the instructions of the assay kit.

[0067] The kits used for oxidative stress and inflammatory factor assays are as follows: Reduced glutathione (GSH) assay kit (microplate method) (A006-2-1, Nanjing Jiancheng, Nanjing, China); Malondialdehyde (MDA) assay kit (TBA method) (A003-1-1, Nanjing Jiancheng, Nanjing, China); Mouse IL-6 ELISA Kit (KE10007, PTG, China); Mouse IL-1 beta ELISA Kit (KE10003, PTG, China); Mouse TNF-alpha ELISA Kit (KE10002, PTG, China). All assays were performed strictly according to the instructions.

[0068] 5. Serum sex hormone level detection Twenty-one days after the start of the experiment, blood was collected from the eyeballs, allowed to stand at room temperature for about 1-2 hours, centrifuged at 12,000 rpm, and the supernatant, i.e., serum, was collected and stored at -80℃. Subsequently, the levels of E2, Testosterone, and AMH hormones in mice in each group were measured strictly according to the instructions of the assay kit.

[0069] The reagent kits used for serum sex hormone level detection are as follows: Testosterone (E-OSEL-M0003, Elabscience, China); Mouse AMH (Anti-Mullerian Hormone) ELISA Kit (E-EL-M3015, Elabscience, China); QuicKey Pro Mouse E2 (Estradiol) ELISA Kit (E-OSEL-M0008, Elabscience, China).

[0070] 6. Statistical processing methods Data were summarized using Excel spreadsheets, and statistical analysis and graphing were performed using GraphPad 8.0.2 software. Independent samples t-tests were used to compare the two groups. A p-value < 0.05 was considered statistically significant.

[0071] II. Experimental Results 1. Analysis of sex hormone (E2, AMH, Testosterone) levels Reproductive endocrine disorders are the cornerstone of PCOS diagnosis. Figures 2-4 Hormonal changes in the model indirectly reflect its reproductive phenotype.

[0072] E2 (estradiol): Serum E2 levels in the model group of female mice were significantly lower than those in the blank control group (P<0.05). In PCOS, although there are many follicles, development arrests at the small antral follicle stage, and the selection of dominant follicles is impaired, resulting in a relative shortage of mature granulosa cells that synthesize E2. Overall E2 levels may be low or exhibit unstable fluctuations. Low E2 status may be associated with anovulation and abnormal endometrial receptivity.

[0073] AMH (Anti-Müllerian Hormone): Serum AMH levels were significantly elevated in female mice in the model group (P<0.05). This is a very characteristic change in PCOS. AMH is mainly secreted by the granulosa cells of small and medium-sized antral follicles. The number of such follicles in the ovaries of PCOS patients is significantly increased, leading to excessive AMH production. High AMH inhibits FSH sensitivity, further hindering the selection and ovulation of dominant follicles, forming a vicious cycle.

[0074] Testosterone: Serum testosterone levels were significantly elevated in female mice in the model group (P<0.05), suggesting the possible presence of hyperandrogenemia in the female model. This elevated testosterone level in female mice may indicate letrozole-induced liver injury or systemic stress affecting ovarian function. However, in typical female PCOS models, the focus is on hyperandrogenemia. Combined with the background of elevated inflammatory factors (such as TNF-α inhibiting aromatase) and AMH, it can be inferred that elevated free testosterone or androstenedione is a direct cause of ovulation disorders in female PCOS models.

[0075] Chicken eye grass extract treatment had a positive regulatory effect on the sex hormone spectrum: it significantly increased the E2 level in model female mice (P<0.05), restoring it to the normal range, suggesting that it may promote further follicular development and granulosa cell function. Figure 2 Simultaneously, it significantly reduced abnormally elevated AMH levels (P<0.05), which may mean that *Centella asiatica* helps reduce the number of arrested follicles or inhibits excessive AMH secretion by their granulosa cells, thereby breaking the inhibition of follicle maturation. Figure 3 Regarding androgens, *Centella asiatica* extract showed a tendency to protect or restore testosterone levels in female mice, reducing excessively high androgen levels. Figure 4 These results indicate that *Lysimachia christinae* extract can improve the core reproductive endocrine disorders of PCOS.

[0076] 2. Analysis of oxidative stress indicators (MDA, GSH) Oxidative stress and the pathogenesis of PCOS mutually promote each other. Excessive production of reactive oxygen species (ROS) can damage oocyte quality, granulosa cell function, and aggravate insulin resistance.

[0077] MDA (malondialdehyde): As the end product of lipid peroxidation, its level directly reflects the degree of oxidative damage in tissues. The MDA content in liver tissue homogenate of the model group was significantly increased (P<0.05), indicating severe oxidative damage in PCOS model mice. Excessive MDA can attack cell membranes, proteins, and DNA, further impairing the normal function of the ovaries and metabolic tissues.

[0078] Glutathione (GSH) is the most important non-enzymatic antioxidant in cells. The significantly reduced GSH levels in the model group (P<0.05) indicate a depletion of the body's antioxidant defense system and a severe deficiency in its ability to scavenge reactive oxygen species (ROS).

[0079] Chicken eye grass extract treatment reversed this imbalance: such as Figure 5 and Figure 6As shown, the high-dose group of *Centella asiatica* extract significantly reduced the MDA content in liver tissue homogenate of model mice (P<0.05) while significantly increasing GSH levels (P<0.05). This indicates that *Centella asiatica* extract can protect ovarian reserve function, improve the follicular development environment, and alleviate oxidative stress-related insulin resistance.

[0080] 3. Analysis of inflammatory cytokine (IL-6, TNF-α, IL-1β) levels Chronic low-grade inflammation is one of the core characteristics of PCOS, and it is involved in the development of insulin resistance, follicular development disorders, and long-term cardiovascular risks.

[0081] IL-6 (interleukin-6): The level of IL-6 in the liver tissue homogenate of the model group was significantly increased (P<0.0001 or P<0.01), indicating that the body was in a strong pro-inflammatory state. IL-6 can be secreted by adipose tissue, immune cells, etc., and can directly induce the liver to produce C-reactive protein, exacerbate systemic insulin resistance, interfere with the function of ovarian granulosa cells, and affect follicle maturation.

[0082] TNF-α (tumor necrosis factor-α): The TNF-α level in the model group was also significantly higher than that in the control group (P<0.05). TNF-α is another key pro-inflammatory cytokine that exacerbates insulin resistance by interfering with the phosphorylation of insulin receptor substrates and can directly inhibit ovarian aromatase activity, leading to impaired conversion of androgens to estrogens, thereby maintaining a hyperandrogenic environment.

[0083] IL-1β (interleukin-1β): IL-1β levels were significantly elevated in the model group (P<0.05). IL-1β plays an important role in local ovarian inflammation, disrupting the follicular microenvironment, promoting follicular atresia, and is closely related to insulin resistance.

[0084] like Figure 7 , Figure 8 and Figure 9 As shown, after intervention with *Centella asiatica* extract, the levels of the three pro-inflammatory cytokines mentioned above were significantly reduced (P<0.05 compared with the model group). This effect indicates that *Centella asiatica* extract can effectively inhibit systemic and potential local ovarian inflammatory responses in the PCOS model, and the reduction of inflammation is an important basis for improving insulin resistance and ovarian function.

[0085] 4. Analysis of the appearance and pathological sections of mouse ovaries like Figure 10As shown, in terms of appearance, the ovaries of the model group (PCOS) mice were generally significantly larger than those of the blank control group, with a pale, smooth surface and visible translucent cystic protrusions, presenting a typical "polycystic" appearance. After intervention with *Centella asiatica* extract, the ovarian enlargement and cystic protrusions on the surface of mice in each treatment group (PCOS+KsL, PCOS+KsM, PCOS+KsH) improved in a dose-dependent manner. Among them, the ovaries of the high-dose group (PCOS+KsH) had the appearance closest to that of the blank control group.

[0086] The control group showed normal ovarian structure, with follicles at different developmental stages (primordial, growing, and mature follicles) and corpora lutea. Follicular layers were clearly defined, granulosa cells were tightly packed, and there was no abnormal stroma proliferation. The model group, however, exhibited classic PCOS pathological features: significantly thickened ovarian cortex filled with numerous cystic, dilated antral follicles. These follicles had thin walls, were filled with fluid, and lacked mature cumulus-oocyte complexes. A key feature was the rare or complete absence of corpora lutea, confirming ovulatory failure. Simultaneously, the theca cell layer often showed varying degrees of proliferation. In the *Corneye Grass* treatment group (PCOS+KsL / M / H): Pathological changes were significantly reversed, showing dose-dependent improvement. In the low-dose group (PCOS+KsL): The number of cystic follicles in the cortical area remained relatively high, but a small number of more mature follicular structures were visible. In the medium- and high-dose groups (PCOS+KsM / H): Improvement was particularly significant. The number of cystic follicles accumulated in the cortical area was significantly reduced, and more normally shaped growing follicles appeared. The most important finding was the observation of newly formed corpora lutea in ovarian sections from the high-dose group (PCOS+KsH) and the medium-dose group. The presence of corpora lutea provides direct morphological evidence for ovulation and the recovery of ovarian cycle function. Furthermore, the degree of theca cell proliferation was also reduced compared to the model group.

[0087] The metabolism-inflammation-oxidative stress axis: The model group showed a self-reinforcing vicious cycle of weight gain, a surge in inflammatory factors (IL-6, TNF-α, IL-1β), and worsening of oxidative stress markers (MDA↑, GSH↓). Obesity / insulin resistance promotes inflammation and oxidative stress; inflammatory factors (such as TNF-α) directly exacerbate insulin resistance; oxidative stress, in turn, damages the insulin signaling pathway and aggravates inflammation. *Chicken Eye Grass* extract effectively disrupts this cycle by simultaneously reducing inflammation (lowering IL-6, TNF-α, IL-1β) and increasing antioxidant activity (lowering MDA, increasing GSH), which explains its improvement in metabolic indicators (such as weight).

[0088] The inflammation / stress-ovarian axis: Elevated inflammatory factors and oxidative stress products directly harm the ovaries. TNF-α and IL-1β can interfere with gonadotropin signaling, induce granulosa cell apoptosis, and inhibit E2 synthesis. Oxidative stress damages the mitochondrial function of oocytes and granulosa cells. These factors collectively lead to follicular development disorders (manifested as abnormal E2 and elevated AMH) and anovulation. *Centella asiatica* extract creates conditions for normal follicular development by eliminating these harmful environments, as reflected in the recovery of E2 and the decrease of AMH.

[0089] Causes and improvement of hyperandrogenemia: In the model, inflammation (especially TNF-α) and possible insulin resistance can stimulate excessive androgen production by ovarian theca cells through different mechanisms and inhibit its conversion to E2 in granulosa cells (aromatase inhibition). The potent anti-inflammatory effect of *Centella asiatica* extract, and its indirect improvement of insulin sensitivity by reducing oxidative stress, may be the reason for its prevention or pre-lowering of androgen levels.

[0090] In summary, the modeling method of this invention successfully induced a complex phenotype similar to PCOS, including metabolic abnormalities (weight gain), a significant chronic inflammatory state, severe oxidative stress damage, and characteristic reproductive endocrine disorders (decreased E2, increased AMH). *Centella asiatica* extract, especially the high-dose group, effectively inhibited systemic inflammation, antagonized oxidative stress, and ultimately positively regulated sex hormone balance (increasing E2 and decreasing AMH). These results strongly suggest that the mechanism of *Centella asiatica* extract in treating PCOS is not singular, but rather involves a comprehensive improvement of the pathophysiological basis of PCOS through its core anti-inflammatory, antioxidant, and metabolic regulatory effects, thereby restoring reproductive endocrine homeostasis.

[0091] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. The use of *Chicken Eye Grass* extract in the preparation of drugs for the prevention and / or treatment of polycystic ovary syndrome.

2. Application of *Chicken Eye Grass* extract in combination with hormone drugs, insulin sensitizers and / or ovulation-inducing drugs in the preparation of drugs for the prevention and / or treatment of polycystic ovary syndrome.

3. The application as described in claim 1 or 2, characterized in that, The extract of *Centipeda minima* includes *Centipeda minima* aqueous extract, *Centipeda minima* alcoholic extract, and *Centipeda minima* aqueous-alcoholic extract.

4. The application as described in claim 3, characterized in that, The extract of *Chicken Eye Grass* is an aqueous extract of *Chicken Eye Grass*. Preferably, the water extract of *Chicken Eye Grass* is prepared by the following method: *Chicken Eye Grass* is taken, crushed to obtain coarse powder, then water is added and refluxed for extraction 1 to 3 times, the filtrates are combined, concentrated, and dried to obtain the extract.

5. The application as described in claim 4, characterized in that, The mass ratio of the coarse powder to water is 1:(8~12); and / or, The reflux extraction temperature is 110℃~130℃, and the time is 1h~2h.

6. The application as described in claim 1, characterized in that, The applications include regulating hormone levels associated with polycystic ovary syndrome; Preferably, this includes increasing estradiol levels, decreasing anti-Müllerian hormone levels, and / or decreasing testosterone levels.

7. The application as described in claim 1, characterized in that, The applications include improving polycystic ovary syndrome-related oxidative stress levels and metabolic abnormalities; Preferably, this includes reducing malondialdehyde levels, increasing glutathione levels, and / or reducing body weight.

8. The application as described in claim 1, characterized in that, The application includes suppressing chronic inflammation associated with polycystic ovary syndrome; Preferably, it includes inhibiting the expression levels of pro-inflammatory factors TNF-α, IL-6 and / or IL-1β.

9. The application as described in claim 1, characterized in that, The applications include improving the clinical symptoms of polycystic ovary syndrome; Preferably, it includes improving oligomenorrhea, amenorrhea, irregular bleeding, hirsutism, acne, ovulation disorders and / or polycystic ovarian changes.

10. The application as described in any one of claims 1 to 9, characterized in that, The dosage forms of the drug include oral and injectable formulations; Preferably, the oral preparation includes tablets, capsules, granules, solutions, powders, emulsions, suspensions, decoctions, pills, and ointments.

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