Bushen Huatan formula improves reproductive and endocrine metabolic disorders in non-obese polycystic ovary syndrome rats via adipose tissue-derived exosomal miR-27a-3p/PPARG signaling pathway

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.

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

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.

Involvement of Kisspeptin in Androgen-Induced Hypothalamic Endoplasmic Reticulum Stress and Its Rescuing Effect in PCOS Rats

Involvement of kisspeptin in androgen-induced hypothalamic endoplasmic reticulum stress and its rescuing effect in PCOS rats

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.

ObjectiveTo identify the association between the phosphorylated Janus kinase 2/phosphorylated signal transducer and activator of transcription (p-JAK2/p-STAT3) signaling pathway and follicular development in polycystic ovary syndrome (PCOS) rats, and explore the underlying mechanism. To evaluate the role of exogenous JAK2 inhibitor AG490 in the model and the associations among luteinizing hormone/choriogonadotropin receptor (LHCGR), follicle-stimulating hormone receptor (FSHR), cytochrome P450 17α (CYP17a), cytochrome P450 19 (CYP19), and PCOS. ResultsRat models of PCOS was established. PCOS rats were intraperitoneally treated with double-distilled water (ddH2O)/DMSO/AG490. The rate of ovarian morphological recovery in the AG490 group was significantly higher compared with the DMSO group (83.3 % vs 9.1 %, X2 = 12.68, P < 0.001). Moreover, the short in the time the estrous cycle was resumed in the AG490 group (hazard ratio = 16.32, P < 0.001) compared with the DMSO group. Compared with the controls, p-JAK2, p-STAT3, LHCGR, and CYP17a expression levels were increased whereas that of FSHR and CYP19 were decreased in the ovaries of PCOS rats. However, an opposite trend was observed after treatment with AG490. Software prediction revealed that the p-STAT3 bound to the promoter regions of LHCGR, FSHR, CYP17a, and CYP19 genes. This finding was confirmed by results of correlation analysis (R = 0.834, −0.836, 0.875 and −0.712, respectively, all P < 0.001). ConclusionThis study demonstrated that the p-JAK2/p-STAT3 signaling pathway was involved in follicular development in PCOS rats by upregulating LHCGR and CYP17a expression, and downregulating that of FSHR and CYP19. AG490 treatment exerted beneficial effects. LHCGR, FSHR, CYP17a, and CYP19 are candidate genes associated with follicular development in PCOS rats.

Therapeutic exploration of polyherbal formulation against letrozole induced PCOS rats: A mechanistic approach

Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder affecting women, often associated with changes in emotional well-being, oxidative stress, and epigenetic modifications. While lifestyle changes are the primary treatment, their roles in ovarian epigenetics in PCOS remain unknown. This study investigates the impact of treadmill exercise and alternate-day feeding (ADF), either individually or combined, on PCOS characteristics and their underlying epigenetic mechanisms in PCOS rats. Female Wistar rats were assigned to either a letrozole-induced PCOS group (n = 30) or a carboxymethyl cellulose (CMC) group (n = 6). Following PCOS confirmation in 6 rats, the remaining PCOS rats were divided into four groups (n = 6/group): control, treadmill exercise, ADF, and combined. After 55 days, vaginal cytology, behavioral assessments, glucose tolerance tests, blood collections for oxidative stress markers, and ovary collection for histological and molecular assessments, were performed. The PCOS rats exhibited irregular estrus, polycystic ovaries, and glucose intolerance. Treadmill exercise and ADF, alone or combined, reduced anxiety-like behavior: rearing (P = 0.024, P = 0.007, P = 0.27), closed arm time (P = 0.047, P = 0.004, P = 0.001), and increased open arm entries (P = 0.034, P = 0.003, P = 0.024). Glucose intolerance was reduced significantly in all treated rats (P = 0.001), with a significantly lower area under the curve in the combined group (P = 0.017). All treated animals showed reduced weight gain and malondialdehyde, significantly in the combined treatment rats (P = 0.001 and P = 0.035). Exercise and ADF, either alone or combined, decreased atretic (P < 0.001, P = 0.005, P = 0.002) and cystic follicles (P = 0.016, P = 0.029, P = 0.016), while increasing corpus luteum (P = 0.002, P = 0.02, P = 0.02). Moreover, the expression of DNMT1 (P = 0.037) and DNMT3B (P = 0.001, and 0.01) decreased in the PCOS + E and PCOS + E + ADF groups, while DNMT3A (P = 0.037) increased in all treated rats, highest in the PCOS + ADF group. These data showed that treadmill exercise and ADF could improve PCOS rat outcomes, possibly by epigenetically modulating gene expression via chromatin modifier enzyme changes. These findings offer a promising avenue for developing a potential epigenetic therapeutic method using chromatin modifiers to reverse PCOS dysregulation.

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.

BackgroundThis study aimed to compare the characteristics of the gut microbiota and their metabolite profiles between polycystic ovary syndrome (PCOS) and orlistat-treated PCOS rats (ORL-PCOS), which could help to better understand the underlying mechanism of the effect of orlistat on PCOS.MethodsPCOS rat models were established using letrozole combined with a high-fat diet. Ten rats were randomly selected as a PCOS control group (PCOS). The other three groups (n = 10/group) were additionally supplemented with different doses of orlistat (low, medium, high). Then, fecal samples of the PCOS and ORL-PCOS groups were analysed by 16S rRNA gene sequencing and untargeted metabolomics. Blood samples were collected to detect serum sex hormones and lipids.ResultsThe results showed that orlistat attenuated the body weight gain, decreased the levels of T, LH, the LH/FSH ratio, TC, TG and LDL-C; increased the level of E2; and improved estrous cycle disorder in PCOS rats. The bacterial richness and diversity of the gut microbiota in the ORL-PCOS group were higher than those in the PCOS group. The ratio of Firmicutes to Bacteroidetes was decreased with orlistat treatment. Moreover, orlistat treatment led to a significant decrease in the relative abundance of Ruminococcaceae and Lactobacillaceae, and increases in the abundances of Muribaculaceae and Bacteroidaceae. Metabolic analysis identified 216 differential fecal metabolites in total and 6 enriched KEGG pathways between the two groups, including steroid hormone biosynthesis, neuroactive ligand-receptor interaction and vitamin digestion and absorption. Steroid hormone biosynthesis was the pathway with the most significant enrichment. The correlations between the gut microbiota and differential metabolites were calculated, which may provide a basis for understanding the composition and function of microbial communities.ConclusionsOur data suggested that orlistat exerts a PCOS treatment effect, which may be mediated by modifying the structure and composition of the gut microbiota, as well as the metabolite profiles of PCOS rats.

Polycystic ovarian syndrome (PCOS) is one of the common causes or female infertility. Phyllanthus muellerianus (Euphorbiaceae) is a plant used to treat various ailments including frequent menstruation and anovulation. We investigated the effects of P. muellerianus extracts on estrus cyclicity, lipid profile, oxidative stress-related markers, sex hormones, and ovarian architecture in letrozole-induced PCOS in rats. After induction of PCOS using letrozole (1 mg/kg/day), normal (n=6), and PCOS (n=108; distributed into 18 groups of 6 animals/group) rats were treated orally for 7 or 14 days with distilled water (10 ml/kg/day), clomiphene citrate (2 mg/kg/day), metformin (500 mg/kg/day), and aqueous or methanolic extract of P. muellerianus (30, 60, and 120 mg/kg). Estrus cyclicity, body, and sexual organ (ovaries and uterus) weights, biochemical and histological parameters were measured. There were letrozole-induced PCOS characterized by irregular estrus cyclicity, elevated (p<0.05-0.01) glycaemia, ovarian weight, triglycerides, total cholesterol, LDL cholesterol, VLDL cholesterol, malondialdehyde, luteinizing hormone (LH), and testosterone concentrations, but there were low (p<0.05-0.001) HDL cholesterol, estradiol, progesterone, catalase, peroxidase, and superoxide dismutase levels, compared with control. PCOS rats had multiple cysts compared with control. These reproductive, biochemical, and structural alterations were alleviated by P. muellerianus extracts. For instance, P. muellerianus restored the estrus cyclicity with a remarkable effect after 14 days of treatment. Moreover, P. muellerianus significantly decreased (p<0.001) LH and testosterone (both extracts; 30, 60, and 120 mg/kg) levels, but increased (p<0.01) estradiol (aqueous extract; 60 mg/kg) concentration. Cystic follicles were also decreased after plant application. P. muellerianus alleviated reproductive, hormonal, and structural alterations in PCOS rats. This plant could be useful in the management/treatment of reproductive and metabolic disorders related to PCOS.

Polycystic ovary syndrome (PCOS) is a common endocrinopathy in women of reproductive age leading to infertility. Besides reproductive and hormonal disturbances, PCOS is often characterized by vitamin D 3 (VD) deficiency. This study aimed to determine the effect of VD on uterine histoarchitecture, the biochemical composition of gland secretions, and the insulin signal transduction pathway using a PCOS rat model. The experiment was conducted on four animal groups (n=8/group): control (C), VD supplemented (VD; 500 IU/day), letrozole-treated (PCOS; 1 mg/kg body weight), and VD-treated PCOS (PCOS+VD) group. Herein, VD supplementation did not improve histomorphometric parameters in the PCOS uterus, whereas clearly influenced sugar composition in uterine gland secretions, restoring their content to that observed in the C group. Furthermore, we found that VD can reduce peripheral and local uterine insulin resistance developed in the PCOS rats via activation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway and subsequent glucose metabolism in the uterus. To sum up, the present results suggest a possible beneficial role of VD supplementation for the maintenance of uterus functions in PCOS rats.

Polycystic ovarian syndrome (PCOS) is an endocrine disorder in women's reproductive age. Currently, the pathophysiology of PCOS is unclear, and the limited treatment options are unsatisfactory. Virgin coconut oil (VCO) is functional food oil associated with pharmacological effects in reproductive disorders. Therefore, we aimed to evaluate whether VCO could enhance clomiphene (CLO) therapy against PCOS in female rats. Rats were randomly divided: (1) Control, (2) PCOS model, (3) PCOS + CLO, (4) PCOS + VCO, and (5) PCOS + CLO + VCO. The PCOS was induced via daily letrozole (1 mg/kg, orally) administration for 21 days. After the PCOS induction, CLO, VCO, and CLO + VCO were administered from days 22 to 36. Serum levels of gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, estrogen, progesterone, and prolactin were estimated. Polymerase chain reaction gene expression for nuclear factor-erythroid-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), catalase (CAT), glutathione reductase (GSR), LH receptor (LHr), androgen receptor (AR), tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and caspase-3 were analyzed. The letrozole-induced PCOS caused considerable increases in GnRH, LH, prolactin, estrogen, and testosterone, whereas FSH decreased significantly compared to the control. The gene expression of Nrf2, HO-1, CAT, and GSR were markedly diminished, while IL-1β, TNF-α, caspase-3, AR, and LHr prominently increased compared to control. Interestingly, the CLO and VCO separately exerted anti-inflammatory and endocrine balance effects. However, VCO-enhanced CLO effect in LH, prolactin and testosterone, Nrf2, HO-1, CAT, GSR, and AR. VCO may synergize with CLO to depress hyperandrogenism and oxidative inflammation in PCOS.

Quercetin modulates ovarian autophagy–related molecules and stereological parameters in a rat model of PCOS