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

Objective To explore the role of isorhamnetin on polycystic ovary syndrome (PCOS) in rats. Methods Sprague Dawley (SD) rats were subcutaneously injected with dehydroepiandrosteron (DHEA) to establish PCOS model. Hematoxylin and eosin (H&E) staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to measure histological changes and apoptosis of ovary tissues. The levels of serum hormones and inflammatory factors in ovary tissues were measured by enzyme-linked immuno sorbent assay (ELISA). Results In DHEA-induced PCOS rats, the levels of serum glucose, insulin, testosterone and luteinizing hormone (LH) were enhanced, estradiol (E2), sex hormone-binding globulin (SHBG), follicle stimulating hormone (FSH) levels were decreased, inflammatory levels and apoptosis of ovary tissues were increased. Additionally, DHEA increased the body weight, ovary weight, and ovary volume, cystic follicles, and decreased corpus luteum. Moreover, the tumor necrosis factor (TNF) signaling pathway was activated in PCOS rats. The levels of TNF receptor superfamily member 1 A (TNFR1), TNF-α, and fas cell surface death feceptor (FAS) were enhanced in ovary tissues of DHEA induced PCOS rats. Isorhamnetin (ISO) treatment after DHEA modeling markedly reduced serum levels of glucose, insulin, testosterone and LH, increased E2, SHBG, FSH level, decreased inflammatory levels, and inhibited apoptosis and decreased body weight, ovary weight, and ovary volume. The levels of TNFR1, TNF-α, and FAS were markedly decreased after ISO treatment in PCOS rats. Additionally, ISO alone had no significant effect on rats. Conclusion Isorhamnetin inhibits inflammatory response to alleviate DHEA-induced PCOS in rats by inactivating the TNF signaling 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 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.

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.

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.

Protective effects of electroacupuncture on polycystic ovary syndrome in rats: Down-regulating Alas2 to inhibit apoptosis, oxidative stress, and mitochondrial dysfunction in ovarian granulosa cells

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

Blood volatile organic compounds as potential biomarkers for poly cystic ovarian syndrome (PCOS): An animal study in the PCOS rat model

The effect of dapsone in testosterone enanthate-induced polycystic ovary syndrome in rat

Objective: To investigate the potential activity of protocatechuic acid in female Wistar rats with letrozole-induced polycystic ovary syndrome (PCOS). Methods: Thirty rats were divided into five groups of six each. Group 1 received 0.5% carboxy methyl cellulose orally and served as the normal control group; group 2 was treated orally with 1 mg/kg of letrozole daily for 21 days and served as the PCOS induced group; group 3 was orally administered with letrozole of 1 mg/kg for 21 days and further administered with standard drug of clomiphene citrate at a dose of 1 mg/kg body weight in 0.5% carboxy methyl cellulose per oral and served as the standard group; groups 4 and 5 were administered with letrozole of 1 mg/kg for 21 days and further treated with protocatechuic acid orally at low dose of 5 mg/kg body weight and high dose of 15 mg/kg body weight respectively for 15 days. At the end of the study period, rats were subjected for the estimation of invasive blood pressure and heart rate, biochemical estimations and antioxidant assay. In addition, ovarian histomorphology was examined. Results: The PCOS was confirmed in the letrozole induced rats with increased concentration of androgen, abnormal lipid levels, glucose, glycosylated haemoglobin and also depletion of antioxidants. After protocatechuic acid treatment, the increased levels of testosterone due to induction of PCOS were restored to normal levels. Additionally, there was a consistent decrease in luteinizing hormone and follicle stimulating hormone levels in the treatment groups, followed by decrease in the number of cysts after treatment with protocatechuic acid. Histopathological observations showed a remarkable recovery of the ovarian tissue and the presence of normalized structure of antral follicle. Protocatechuic acid treatment restored all the parameters to normalcy and abolished cysts formation in ovaries of female rats. Conclusions: Protocatechuic acid shows potential protective effects in letrozole-induced PCOS rats. The protective effect is comparable to that of clomiphene citrate and thus shows its potential in the treatment of PCOS.

To explore the expression level of microRNA-409 in PCOS (polycystic ovary syndrome) rats, as well as its potential effects on fertility of PCOS rats and phenotypes of offspring rats. PCOS model in rats was established by Letasazole administration. Follicular development of rats was evaluated by the percentages of the cystic follicle (FC) and corpus luteum (CL) of all follicles. The enzyme-linked immunosorbent assay (ELISA) was conducted to detect serum levels of hormones in rats, including LH, LH/FSH, T, INS, FSH, and E2. Subsequently, PCOS rats received a subcapsular injection of microRNA-409 mimics. The expression level of microRNA-409 in ovary was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Serum levels of LH, LH/FSH, T, INS, FSH, and E2 in PCOS rats with microRNA-409 overexpression were accessed by enzyme-linked immunosorbent assay (ELISA) as well. PCOS rats were mated with male rats for recording pregnancy rate. At 6-week-old of offspring, they were sacrificed for detecting microRNA-409 level, percentages of FC and CL, as well as serum levels of hormones. PCOS rats showed irregular estrous cycle and they were mainly in the anestrum. Rats in the control group were in a regular estrous cycle. A higher percentage of FC and a lower percentage of CL were seen in PCOS rats compared with those of controls. ELISA data revealed higher serum levels of LH, LH/FSH, and T in PCOS rats compared with those of controls. However, levels of FSH and E2 were lower in PCOS rats. Although INS level increased in PCOS rats, we did not observe a significant difference in INS level between PCOS rats and control rats. MicroRNA-409 was lowly expressed in ovaries of PCOS rats than those of controls. After injection of microRNA-409 mimics into rat ovary, microRNA-409 expression remarkably upregulated than those PCOS rats without injection. Rats in PCOS+microRNA-409 mimics group showed the largest body weight compared with those in the PCOS group and control group. PCOS rats showed a lower pregnancy rate than those of controls, which was markedly increased after administration of microRNA-409 mimics. Rats in PCOS+microRNA-409 mimics group presented lower levels of LH, LH/FSH, T, and INS, but higher levels of FSH and E2 than those in PCOS group. MicroRNA-409 is lowly expressed in the ovary of PCOS rats. Overexpression of microRNA-409 could improve hormone levels and pregnancy rate in PCOS rats, as well as affect clinical phenotypes of their offspring.