Isorhamnetin inhibits inflammatory response to alleviate DHEA-induced polycystic ovary syndrome in rats

Objective:The effect of trans-anethole and metformin on biochemical and hormonal changes of testosterone-induced Polycystic ovary syndrome (PCOS) in rats was investigated.Materials and Methods:Female Wister rats (n=48) were randomly divided into six groups: control; PCOS; PCOS+metformin (300 mg/kg); and PCOS+trans-anethole (20, 40, and 80 mg/kg). PCOS was induced by intraperitoneal injection of testosterone (1 mg/kg/day) for 35 days. After induction of PCOS, trans-anethole and metformin were given orally for 30 days. Finally, blood sugar, insulin, lipid profile, and testosterone and dehydroepiandrosterone (DHEAS) as well as animals’ weight, and water and food intake were determined.Results:In all treated and untreated PCOS groups, serum testosterone levels were significantly increased compared to the control group (p<0.001 for all groups). Treatment of rats with trans-anethole or metformin significantly reduced serum levels of cholesterol, insulin, triglycerides, testosterone and DHEAS (only in PCOS+trans-anethole groups) compared to the PCOS group (p<0.01-p<0.001). Weight gain in the PCOS animals increased significantly compared to the control group (p<0.001), while in the metformin- and trans-anethole (40 and 80)-treated animals it decreased significantly compared to the PCOS group (p<0.01-p<0.001).Conclusion:These results showed that trans-anethole significantly decreased serum levels of insulin, DHEAS and blood lipids. It can be concluded that trans-anethole ameliorates PCOS biochemical and hormonal change in PCOS rats; therefore, it might be suggested as a beneficial remedy for further clinical evaluations in PCOS patients.

This study aims to explore the therapeutic effect of Yuzhi Zhixue Granules on polycystic ovary syndrome(PCOS) in rats and explain the underlying mechanism by metabolomics. Rats were randomized into normal, model, low-, medium-, and high-dose(0.5, 1.5, and 4.5 g·kg~(-1)·d~(-1)) Yuzhi Zhixue Granules, and positive control(metformin, 0.2 g·kg~(-1)·d~(-1)) groups. The rats in other groups except the normal group were administrated with 1 mg·kg~(-1)·d~(-1) letrozole and fed with a high-sugar and high-fat diet for the modeling of PCOS. After 40 days of modeling, the normal and model groups received distilled water and letrozole+distilled water, respectively, and other groups received letrozole and corresponding drugs, once a day for 50 days. The oral glucose tolerance test(OGTT) was carried out and enzyme-linked immunosorbent assay(ELISA) was conducted to detect insulin release, and the radioimmunoassay was employed to measure the serum levels of follicle-stimulating hormone(FSH), luteinizing hormone(LH), estradiol(E_2), and testosterone(T). The serum levels of high density lipoprotein-cholesterol(HDL-C), low density lipoprotein-cholesterol(LDL-C), and triglyceride(TG) were determined by an automatic biochemical analyzer. The pathological changes in the ovary were observed by hematoxylin-eosin(HE) staining. The protein levels of phosphatidylinositol 3-kinase(PI3K), protein kinase B(Akt), and their phosphorylated forms in the ovary were determined by Western blot. Ultra-high performance liquid chromatography-tandem mass spectrometry(LC-MS/MS) was employed to study the fecal and serum metabolites in the rat model of PCOS. The results showed that compared with the model group, drug administration repaired the impaired glucose tolerance, enhanced the insulin sensitivity, elevated the serum levels of HDL-C and E_2, lowered the serum levels of TG and T, ameliorated the pathological changes in the ovarian tissue, and up-regulated the protein levels of p-PI3K and p-Akt in the ovarian tissue. A total of 46 differential metabolites and 10 metabolic pathways in the fecal samples were screened out, which were mainly related to the biosynthesis of unsaturated fatty acids and tyrosine metabolism. In terms of the serum metabolism, 34 differential metabolites and 15 metabolic pathways were screened out, mainly related to the biosynthesis of unsaturated fatty acids and aminoacyl-tRNA. In conclusion, Yuzhi Zhixue Granules can alleviate the disorders of glucose, lipids, and sex hormones and improve the ovarian status in the rat model of PCOS by regulating the serum and fecal metabolism and activating the PI3K/Akt pathway.

This study investigated the effects of proanthocyanidins (PCs) on ovarian fibrosis in letrozole-induced polycystic ovary syndrome (PCOS) in rats. The administration of PCs effectively reduced the body weight (BW) and relative ovarian weight in PCOS rats. ELISA results revealed that PCs significantly reduced the level of serum T, LH, LH/FSH in the PCOS group. In addition, qRT-PCR results revealed that treatment with PCs significantly increased the main antioxidant enzymes (Cat, Sod2, Gpx3, Mgst1, Gsta4, Sod1 and Prdx3) in PCOS rats. Also, the expression analysis of proteins by Western blotting revealed that PCs significantly decreased the level of TGF-βR1, p-Smad3, p-Smad2 and Smad4 and reversed the downregulation of Smad7 in PCOS rats. The study suggested that PCs improved ovarian fibrosis in PCOS rats by regulating the serum hormone level, inhibiting oxidative stress and suppressing the activation of the TGF-β1/Smads signaling pathway. PRACTICAL APPLICATIONS: Currently, plant extracts are being widely used to treat female reproductive and metabolic disorders. Particularly, proanthocyanidins (PCs), the well-known natural polyphenolic compounds, which are a significant source of antioxidants present in many colored fruits, are consumed as fruits as well as a dietary supplement to prevent many disorders. Recent pharmacological studies have reported that PCs have many health beneficial properties, such as antioxidant activity, improving cholesterol homeostasis, blood lipid regulatory properties, microcirculation improvement effect, antitumor activity and anti-aging activity. Despite these properties of PCs, the antifibrosis effect of PCs has not been studied to date. The main purpose of this study was to research the role and the mechanisms of PCs in ovarian fibrosis in PCOS rats.

This study was undertaken to study the effects of melatonin on metabolic and reproductive aspects of polycystic ovary syndrome (PCOS) in rats. PCOS was induced by daily subcutaneous administration of testosterone (20 mg/kg) to 21-day-old female rats for 35 days. Rats were given metformin (500 mg/kg), melatonin (1 mg/kg) or melatonin (2 mg/kg) along with testosterone. One group served as vehicle control. On the 36th day, the animals were euthanised, and anthropometrical, biochemical (glucose, insulin, lipids, testosterone, C reactive protein (CRP)), oral glucose tolerance test, and histopathological evaluation of ovaries, uterus and intraabdominal fat (IAF), were carried out. Daily colpocytological examination was carried out from 14(th) day of study until termination. Both the doses of melatonin significantly reduced body weight, body mass index, IAF, insulin and CRP. A favourable lipid profile, normal glucose tolerance and a decrease in the percentage of estrus smears were observed. Histopathological examination of ovary, uterus and IAF revealed a decrease in the number of cystic follicles, decrease in neoplastic endometrial glands, and decrease in adipocyte hypertrophy, respectively. The effects observed with melatonin were comparable to that with metformin. The study provides evidence of the potential beneficial effects of melatonin in PCOS.

The study designed to evaluate influence of kisspeptin on polycystic ovary syndrome in female rats, First experiment (Induction of polycystic ovary): thirtieth virgin female rats divided randomly into two groups first group via used ten rats serves as control group . While the second group twenty female rat (PCOS-induce group) were orally administered with letrozole at a dose of 1 mg/kg/21 days. The Second Experiment Effect of treatments with kisspeptin on PCOS : Animals were divided as follows: first group (negative control) .Second group (Positive control) .While animals in third and fourth groups (PCOS+Kisspeptin): five animal in each group PCOS-induced rats administrated of 20 and 40 nmol/rat S/C 21 days Kisspeptin daily respectively .The result show in first experiment a significant increase in serum activity of LH, FSH, estrogen and testosterone and significant decrease progesteron concentration in serum of female rats treated with letrozole. While in the second experiment there was significant decrease in FSH in the C+ compared with C and PCOS treated group. A significant increase in the LH in C+ compared with C and treated group 20 and 40 nmol/rat Kisspeptin . Also the results showed that there was a significant decrease in serum estrogen concentration was observed in PCOS nontreated group compared with control. On other hand a significant increase in serum estrogen in T1(20 nmol/kg B.W) compared with T2(40 nmol/kg B.W). The results of the histopathological study of ovary after treatment PCOS with kisspeptin 20 nmol/kg show partial return of follicular cyst to normal ovarian tissue, but the treated with 40 nmol/kg show semicomplete treated and return follicular cyst to normal ovarian tissue. On conclusions, the present study confirmed that PCOS affect the female reproductive hormonal balance and kisspeptin 40 nmol/rat daily S/C injection show effect in normal restoring of female reproductive hormonal and histological balance of PCOS rats.

Lycopene (LYC) is an extremely powerful antioxidant with the potential to treat a range of diseases and to inhibit ferroptosis. This research aims to elucidate how LYC impacts polycystic ovarian syndrome (PCOS) and the action mechanisms. A PCOS rat model was constructed by injecting DHEA. Different doses of LYC were injected intraperitoneally in PCOS rats, the estrous cycle was recorded. The histopathological damage of ovary in PCOS rats was observed by HE staining, testosterone (T), estradiol (E2), luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were examined by ELISA kits. Transmission electron microscopy, prussian blue staining, biochemical kits to determine ferroptosis. Immunohistochemistry and Western blot to assess the levels of ferroptosis-related and AMPK/Nrf2 pathway-related proteins to explore whether LYC affects ferroptosis in PCOS through this pathway. PCOS rats had significantly higher body weights, ovarian weights and ovarian indices, and disorganized estrous cycles, which were dose-dependently ameliorated by LYC. In addition, LYC significantly ameliorated the histopathological damage of ovary in PCOS rats and restored the normal secretion of T, E2, LH, and FSH. LYC attenuates iron deposition in PCOS ovarian tissues, reduces iron and ROS levels, and inhibits ferroptosis. Notably, LYC activated the AMPK/Nrf2 pathway, and AMPK inhibitor intervention attenuated the therapeutic effect of LYC in PCOS rats, suggesting that LYC acts through the AMPK/Nrf2 pathway. LYC attenuates estrous cycle disruption, ameliorates pathological impairments, and inhibits ferroptosis in PCOS rats by modulating the AMPK/Nrf2 pathway.

The relationship between apoptosis of granulosa cells and follicle development arrest in polycystic ovarian syndrome (PCOS) rats, and the contribution of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in apoptosis of granulosa cells were explored. By using sodium prasterone sulfate rat PCOS model was induced. The apoptosis of granulosa cells in ovaries of rats was observed by TdT-mediated dUTP-biotin nick end-labeling (TUNEL), and the expression of TRAIL protein and mRNA in granulosa cells was detected by using immunohistochemical staining and reverse transcription polymerase chain reaction (RT-PCR) respectively. The apoptotic rate and the expression of protein TRAIL in granulosa cells were significantly higher in antral follicles from the PCOS rats than in those from the control rats (P<0.01, P<0.05). There was no significant difference in apoptotic rate and the expression of TRAIL protein in granulosa cells of preantral follicles between the PCOS rats and the control rats (P>0.05). No apoptosis and the expression of TRAIL protein in granulosa cells of primordial follicles were found in the two groups. The expression of TRAIL mRNA was significantly stronger in granulosa cells from the PCOS rats than in those from the control rats (P<0.01). It was suggested that the apoptotic rate in granulosa cells was significantly higher in antral follicle from the PCOS rats than in those from the control rats. TRAIL played a role in regulating the apoptosis of granulosa cells in PCOS rats.

Levels of spexin and its receptors GALR2 and GALR3 in the hypothalamus and ovary of letrozole-induced polycystic ovary syndrome in rats

Soy isoflavones have been widely used in the treatment of clinical gynecological diseases. The aim of this study was to investigate the therapeutic effect and molecular mechanism of Soy isoflavones on rats with polycystic ovary syndrome (PCOS). Sprague-Dawley rats were orally administered 1 mg/kg letrozole for 21 consecutive days to induce the PCOS rat model. After PCOS induction, Soy isoflavones (100 mg/kg) or metformin (Positive control; 500 mg/kg) was administered continuously for 28 days. Then, H&E staining was used to observe the pathological changes of ovary. The serum hormone levels and the levels of antioxidant and inflammatory cytokines in ovarian tissue were detected. Additionally, the expression of NF-κB signaling pathway protein was detected by Western blot. Our results showed that soy isoflavones treatment significantly reduced the body weight, ovarian volume and weight, and improved estrous cycle in PCOS rats. H&E staining showed that the number of cystic dilated follicles and atretic follicles in ovarian tissue diminished, showing healthy follicles and corpus luteum. In addition, soy isoflavones treatment markedly decreased serum testosterone and luteinizing hormone (LH) levels, as well as oxidative stress levels and inflammation levels, and increased estradiol (E2) and follicle stimulating hormone (FSH) levels. At the same time, Soy isoflavones treatment inhibit the phosphorylation level of NF-κB p65 and increased the IκBα expression in ovarian tissues of PCOS rats. Overall, Soy isoflavones can improve ovarian morphology and hormone disorders in PCOS rats by inhibiting the activity of NF-κB pathway and enhancing anti-inflammatory and antioxidant capacity.

Effect of Rhei Radix Et Rhizome on treatment of polycystic ovary syndrome by regulating PI3K/AKT pathway and targeting EGFR/ALB in rats

Brown adipose tissue activation by rutin ameliorates polycystic ovary syndrome in rat

Polycystic ovary syndrome (PCOS) is a common endocrine disease accompanied by energetic metabolic imbalance. Because the etiology of PCOS is complex and remains unclear, there is no effective and specific treatment for PCOS. It is often accompanied by various metabolic disorders such as obesity, insulin resistances, and others. Activated brown adipose tissue (BAT) consumes excess energy via thermogenesis, which has positive effects on energy metabolism. Our previous research and that of others indicates that BAT activity is decreased in PCOS patients, and exogenous BAT transplantation can improve PCOS rodents. Notably however, it is difficult to apply this therapeutic strategy in clinical practice. Therapeutic strategies of enhancing endogenous BAT activity and restoring whole-body endocrine homeostasis may be more meaningful for PCOS treatment. In the current study, the dehydroepiandrosterone-induced PCOS rat was exposed to low temperature for 20 days. The results show that cold treatment could reverse acyclicity of the estrous cycle and reduce circulating testosterone and luteinizing hormone in PCOS rats by activating endogenous BAT. It also significantly reduced the expression of steroidogenic enzymes as well as inflammatory factors in the ovaries of PCOS rats. Histological investigations revealed that cold treatment could significantly reduce ovary cystic follicles and increase corpus luteum, indicating that ovulation was recovered to a normal level. Concordant with these results, cold treatment also improved fertility in PCOS rats. Collectively, these findings suggest that cold treatment could be a novel therapeutic strategy for PCOS.

Objective: To investigate the combined therapeutic potential of Gymnema (G.) sylvestre and Pergularia (P.) daemia on letrozole-induced polycystic ovarian syndrome (PCOS) in rats. Methods: Thirty six healthy female Wistar rats with regular estrus cycles were randomly divided into six groups each of 6. Group I received 1 mL of 0.5% carboxyl methyl cellulose orally and served as the vehicle control group, while groups II to VI were treated with letrozole (1 mg/kg body weight p. o.) for 21 days to induce PCOS. After induction of PCOS, group II served as the PCOS control group, without treatment; group III received metformin (20 mg/kg body weight p. o.) as the standard group, and groups IV to VI received G. sylvestre (100 mg/kg body weight p. o.), P. daemia (300 mg/kg body weight p. o.), and the combination of G. sylvestre and P. daemia, respectively, for 28 days. Vaginal smears were collected from all rats daily throughout the study to determine the phases of the estrus cycle. After completing the treatment schedule, oral glucose tolerance test, serum lipid profile and reproductive hormonal analysis were carried out. Subsequently, the rats were sacrificed to collect ovary and uterus for histopathological examination. Results: The PCOS control rats showed a significant irregularity in the estrus cycle, hyperglycemia, and the altered serum lipid profile such as the increased low and very low density lipoprotein, triglyceride, and decreased high density lipoproteins. In addition, the PCOS control rats showed a significant increase in serum luteinizing hormone, testosterone, and estrogen, and decrease in follicle stimulating hormone and progesterone. These changes were significantly revoked in all the treatment groups. The test drugs also significantly reduced the gained ovary weight (P&lt;0.001), and histopathology of the ovary showed almost normal ovary. Among the treatment groups, the group of combination treatment of G. sylvestre and P. daemia showed superior ameliorative results in PCOS parameters. Conclusions: Combination of G. sylvestre and P. daemia presents potent synergistic activity against hyperandrogenism, hyperinsulinemia, anovulation and follicular cysts in letrozole-induced PCOS rats.

Effect of combination of Withania somnifera Dunal and Tribulus terrestris Linn on letrozole induced polycystic ovarian syndrome in rats

Numerous inducers of polycystic ovarian syndrome (PCOS) at different doses have been proposed in several experimental animals but there is no consensus on an appropriate dose(s) that should ideally reproduce the key biochemical and clinical features of PCOS similar to those of humans. Therefore, this study was aimed at investigating an appropriate dose(s) for the induction of PCOS in female Wistar rats. Twenty-four female albino rats (190.00 ± 13.00 g) with 4-5 days of estrus cyclicity were completely randomized into 4 groups (A - D) of six animals each. Animals in group A (control) were subcutaneously administered 0.2 ml of pure olive oil, while those in groups B, C and D were subcutaneously administered same volume corresponding to 5.0, 7.5 and 10.0 mg of mifepristone in olive oil for 9 days starting from the day of estrus (Day 1). The estrus cycle, serum testosterone (T), estradiol (E), prolactin (Pr), follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), insulin (Is), weight of the animals, fasting blood glucose (FBS) and ovarian morphology were monitored/evaluated/examined. The 5.0 mg of mifepristone extended the estrus stage for four days, increased (p<0.05) the levels of serum E, P, Pr, FSH, T, triacylglycerides (TAG), and total cholesterol (TC) as well as decreased the concentrations of LH and high density lipoprotein-cholesterol (HDL-C). There was no significant difference (p<0.05) in the Is concentration, animal body weights and FBS at day 10 in rats administered 5.0 mg of mifepristone. The 7.5 mg of mifepristone produced irregular estrus cycle, increased Pr, TAG, T, and TC concentrations and FBS whereas it decreased E, P, HDL-C, and LH. The Is, FSH and body weights of the animals were not significantly altered at 7.5 mg of mifepristone. The 10.0 mg of mifepristone produced irregular estrus cycle, increased the levels of E, TAG, Is, LH, T as well as decreased the levels of P and HDL-C. The levels of Pr, FSH, TC, body weights and FBS were not significantly altered at this dose. There was no ovarian follicular growth and atresia in the 5.0 and 7.5 mg mifepristone-treated rats whereas the 10 mg of mifepristone produced these histopathological features. Overall, the study concluded that subcutaneous administration of mifepristone (RU486) induces polycystic ovarian syndrome in rats through deprivation of progesterone with the 10 mg producing majority of the key biochemical and clinical features associated with PCOS in humans. The study, therefore, recommends the subcutaneous administration of mifepristone (RU486) on daily basis for 9 days as a good model for inducing PCOS in animals.