Rat Model Of Polycystic Ovary Syndrome Research Articles

Rubus chingii Hu relieved the polycystic ovary syndrome with enhanced insulin sensitivity through inhibiting TXNIP/NLRP3 inflammasome signaling

Background Polycystic ovary syndrome (PCOS) is one of the most prevalent endocrine disorders in gynecology with severe metabolic abnormalities. Therefore, identifying effective treatments and drugs for PCOS is important. We aimed to investigate effect of the traditional Chinese medicine (TCM) Rubus chingii Hu (R. chingii) on ovarian function and insulin resistance (IR) of PCOS rat models, and to explore the underlying mechanisms Methods A PCOS rat model was established by subcutaneous injection of dehydroepiandrosterone (DHEA) solution for 20 days. PCOS rats were randomly divided into a control group (CON), model group (MOD), metformin group (MET), TCM R. chingii group (RCG), and RCG + Ad-TXNIP groups. After 28 days of treatment, the samples were collected for subsequent experiments. Results R. chingii treatment alleviated hormone imbalance and IR while improving ovarian pathology in the PCOS model. R. chingi inhibited the activation of the thioredoxin-interacting protein (TXNIP)/NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in the ovarian tissue of PCOS rats. Furthermore, TXNIP overexpression hindered the protective effect of R. chingii intervention in PCOS rats, as evidenced by the increase of homeostasis model assessment of insulin resistance (HOMA-IR), luteinizing hormone (LH), testosterone (T), C-reactive protein (CRP) levels, and atretic follicles. Conclusion R. chingii intervention improved ovarian polycystic development by suppressing the TXNIP/NLRP3 inflammasome, which may be an effective treatment for PCOS.

TMT-Based Proteomics Analysis of the Intervention Effect of Orlistat on Polycystic Ovary Syndrome Rats Induced by Letrozole Combined with a High-Fat Diet.

Polycystic ovary syndrome (PCOS) is a complex gynecological endocrine and metabolic disease. Orlistat as a lipase inhibitor may improve the pathological characteristics of PCOS and is the sole antiobesity agent available in various countries. In this study, the PCOS rat models were established using letrozole and high-fat diet. Tandem Mass Tag labeling peptide coupled with liquid chromatography with tandem mass spectrometry (LC-MS/MS) approach was employed to investigate the differentially expressed ovarian proteins (DEPs) in the PCOS and control rats for the effect of PCOS, and in the PCOS and orlistat-treated PCOS rats for the effect of orlistat in PCOS. The orlistat attenuated the body weight gain; decreased the levels of testosterone, luteinizing hormone, a ratio of luteinizing/follicle-stimulating hormones; increased the level of estradiol; and recovered the estrous cycle in PCOS rats. In addition, 795 and 119 DEPs were found in PCOS and orlistat-treated PCOS groups, respectively. Based on the Gene Ontology and Kyoto Encyclopedia of Gene and Genomes pathway analysis of DEPs, orlistat restored the disturbed metabolism of linoleic acid, arachidonic acid, galactose, and glycerolipids, and then improved the chronic inflammation in PCOS rats. This study analyzed the ovarian proteome of orlistat-treated PCOS rats and identified targeted proteins, which explored the pathogenesis of PCOS and the potential effects of orlistat in PCOS rats.

The effect of Daflon on Polycystic Ovary Syndrome Induced by Letrozole in the Rat model

Background: Polycystic ovary syndrome (PCOS) is a condition in which the ovaries produce an abnormal amount of androgens, male sex hormones that are usually present in women in small amounts. The name polycystic ovary syndrome describes the numerous small cysts (fluid-filled sacs) that form in the ovaries. Objective: This study aimed to evaluate the therapeutic role of Daflon versus myoinositol (MI) in improving the reproductive function in the rat model of PCOS. Methods: Oral letrozole(LTZ) combined with a high-fat diet for 21 days was used to induce the PCOS in Wister rats, group 1: 8 female Wister rats received only the vehicle C.M.C (carboxy methyl cellulose) for 21 days, group 2: received LTZ solution orally for 21days on daily bases(induction group) group 3: received LTZ for 21 days for PCOS induction than after that, Daflon 100mg/kg (DAF-H) for the next 21 days, group 4: recived LTA for 21 days for PCOS induction then received Daflon 50mg/kg (DAF-L) for the next 21 days, and group 5: recived LTZ for 21 days for PCOS induction then MI were administered for the following 21 days. Results: The LTZ-induced PCOS rats have lost their estrous cyclicity and become fixed at the diestrus phase, developed, abnormal sex and gonadotrophin hormone serum levels, increased cystic follicles, decreased number of the growing follicles, and very little or no corpora lutea on microscopic examination, which were reversed by the three drugs, DAF-H, DAF-L, and myoinositol.DAF-H and myoinositol were mostly equally effective in improving the reproductive manifestations of the disease. However, DAF-L was effective in returning the estrus cycle regulatory but yet not as effective in improving the condition compared to DAF-H and MI in this LTZ-induced model of PCOS. Conclusion: Daflon had the potential to ameliorate polycystic ovarian syndrome at the dose of 100mg/kg which is statistically comparable with myo-inositol. This property might be attributed to antioxidant activity of Daflon.

Effect of Canagliflozin Alone and in Combination with Metformin on Ovarian Histology of a Polycystic Ovary Syndrome Rat Model

<p><strong>Background and Objective:</strong> Canagliflozin reduces insulin resistance in diabetics and is hypothesized to produce a beneficial effect in polycystic ovarian syndrome (PCOS). Therefore, this study is planned to compare the effects of canagliflozin and metformin alone and in combination on ovarian histology of rat models with letrozole-induced PCOS.</p>
 <p><strong>Methods:</strong> It was a randomized experimental study on Sprague Dawley rats. A total of N = 40 female rats were divided randomly into six groups (A-F). With the exception of normal control group A, rats were given letrozole 1 mg/kg daily for 21 days till PCOS was induced. Group B was disease control, while rats in groups C-F were administered canagliflozin (10 mg/kg), metformin (100 mg/kg), a combination of canagliflozin (10 mg/kg) with metformin (100 mg/kg), and combination of canagliflozin (5 mg/kg) with metformin (50 mg/kg), respectively. Animals were sacrificed on the 48th day. Ovaries and uterus were removed, weighed, and processed for further histopathological analysis.</p>
 <p><strong>Results:</strong> All treatment groups showed significant improvement in ovarian histology. The number of primary and secondary follicles and cystic follicles was significantly lower (p < 0.001) in all treatment groups as compared to the disease control group.</p>
 <p><strong>Conclusion:</strong> Canagliflozin is effective for the treatment of PCOS and augments the effect of metformin in a rat model.</p>

LncRNA PWRN2 promotes polycystic ovary syndrome progression via epigenetically reducing ATRX by recruiting LSD1

Long non-coding RNA has been shown to mediate the progression of polycystic ovary syndrome (PCOS). However, the role and mechanism of Prader-Willi region nonprotein coding RNA 2 (PWRN2) in PCOS progression remain unclear. In our study, Sprague-Dawley rat was injected with dehydroepiandrosterone to mimic PCOS rat models. HE staining was used to assess the number of benign granular cells, and serum insulin and hormone levels were detected by ELISA kit. The expression of PWRN2 was examined by qRT-PCR. Ovarian granulosa cells (GCs) proliferation and apoptosis were examined by CCK-8 assay and flow cytometry. The protein levels of apoptosis markers and Alpha thalassemia retardation syndrome X-linked (ATRX) were determined by western blot. The interaction between lysine-specific demethylase 1 (LSD1) and PWRN2 or ATRX was confirmed by RIP and ChIP assay. Our data showed that PWRN2 was upregulated and ATRX was downregulated in the ovarium tissues and serum of PCOS rat. PWRN2 knockdown promoted GCs proliferation and inhibited apoptosis. In the mechanism, PWRN2 inhibited ATRX transcription by binding with LSD1. In addition, downregulation of ATRX also eliminated the effect of sh-PWRN2 on GCs growth. In conclusion, our data suggested that PWRN2 might restrain GCs growth to promote PCOS progression, which was achieved by binding with LSD1 to inhibit ATRX transcription.

The Effects of Abscisic Acid Administration in PCOS Rat Model and H2O2-Induced Human Granulosa Cell Damage: A Preliminary Investigation

Background: Polycystic ovary syndrome (PCOS) is commonly associated with metabolic disorders. Abscisic acid (ABA), a plant hormone found in vegetables and fruits that can be naturally supplied through dietary intake, has previously been studied for its benefits to human health, particularly in people with diabetes. ABA plays a key role in glucose metabolism, inflammation, and tumor growth. The aim of this study was to investigate the therapeutic effect of ABA on letrozole-induced PCOS rats. Methods: Wistar rats were implanted with continuous-release letrozole pellets to induce a PCOS-like phenotype, and subsequently treated with ABA or vehicle control. Body weight changes, Testosterone (T) levels, fasting insulin measurements, and glucose tolerance tests were investigated. A cell apoptosis model, induced by hydrogen peroxide (H2O2), was established. Cell viability was tested by cell counting Kit-8. Cell apoptosis was tested by flow cytometry, and oxidative stress state of cells was examined via reactive oxygen species (ROS) array. Results: Body weight, glucose impairment, and T level were significantly improved in ABA-treated PCOS rats compared to vehicle-treated rats. ABA significantly reduced H2O2-induced human ovarian granulosa cell line (KGN) cell apoptosis and ROS levels and increased cell viability. Conclusions: The findings suggest that ABA could be an adjunctive treatment candidate for improving the insulin sensitivity and hyperandrogenemia of PCOS patients, and for ameliorating H2O2-induced cell apoptosis and oxidative stress.

AMPK Activation as a Protective Mechanism to Restrain Oxidative Stress in the Insulin-Resistant State in Skeletal Muscle of Rat Model of PCOS Subjected to Postnatal Overfeeding.

Polycystic ovary syndrome (PCOS) is a common endocrinopathy in women of reproductive age, often associated with obesity and insulin resistance. Childhood obesity is an important predisposing factor for the development of PCOS later in life. Being particularly interested in the interplay between prepubertal obesity and hyperandrogenemia, we investigated the effects of early postnatal overfeeding, accomplished by reducing litter size during the period of suckling, on energy sensing and insulin signaling pathways in the gastrocnemius muscle of a rat model of PCOS-induced by 5α-dihydrotestosterone (DHT). The combination of overfeeding and DHT treatment caused hyperinsulinemia and decreased systemic insulin sensitivity. Early postnatal overfeeding induced defects at critical nodes of the insulin signaling pathway in skeletal muscle, which was associated with reduced glucose uptake in the presence of hyperandrogenemia. In this setting, under a combination of overfeeding and DHT treatment, skeletal muscle switched to mitochondrial β-oxidation of fatty acids, resulting in oxidative stress and inflammation that stimulated AMP-activated protein kinase (AMPK) activity and its downstream targets involved in mitochondrial biogenesis and antioxidant protection. Overall, a combination of overfeeding and hyperandrogenemia resulted in a prooxidative and insulin-resistant state in skeletal muscle. This was accompanied by the activation of AMPK, which could represent a potential therapeutic target in insulin-resistant PCOS patients.

MiR-4433a-3p promotes granulosa cell apoptosis by targeting peroxisome proliferator-activated receptor alpha and inducing immune cell infiltration in polycystic ovarian syndrome.

Granulosa cell (GC) proliferation and apoptosis are critical events of the ovum energy supply, which lead to follicular growth retardation or atresia, and various ovulatory obstacles, eventually resulting in the development of ovarian disorders such as polycystic ovarian syndrome (PCOS). Apoptosis and dysregulated miRNA expression in GCs are manifestations of PCOS. miR-4433a-3p has been reported to be involved in apoptosis. However, there is no study reporting the roles of miR-4433a-3p in GC apoptosis and PCOS progression. miR-4433a-3p and peroxisome proliferator-activated receptor alpha (PPAR-α) levels in GCs of PCOS patients or in tissues of a PCOS rat model were examined by quantitative polymerase chain reaction and immunohistochemistry. Bioinformatics analyses and luciferase assays were used to examine the association between miR-4433a-3p and PPAR-α, as well as PPAR-α and immune cell infiltration, in PCOS patients. miR-4433a-3p expression in GCs of PCOS patients was increased. miR-4433a-3p overexpression inhibited the growth of the human granulosa-like tumor cell line (KGN) and promoted apoptosis, while co-treatment with PPAR-α and miR-4433a-3p mimic rescued miR-4433a-3p-induced apoptosis. PPAR-α was a direct target of miR-4433a-3p and its expression was decreased in PCOS patients. PPAR-α expression was also positively correlated with the infiltration of activated CD4+ T cells, eosinophils, B cells, gamma delta T cells, macrophages, and mast cells, but negatively correlated with the infiltration of activated CD8+ T cells, CD56+ bright natural killer cells, immature dendritic cells, monocytes, plasmacytoid dendritic cells, neutrophils, and type 1T helper cells in PCOS patients. The miR-4433a-3p/PPAR-α/immune cell infiltration axis may function as a novel cascade to alter GC apoptosis in PCOS.

Glucagon-like peptide-1 receptor agonists and cardiometabolic risk factors in Polycystic Ovary Syndrome: role of race/ethnicity

Background: Polycystic Ovary Syndrome (PCOS), the most common endocrine disorder in reproductive-age women, is characterized by androgen excess, polycystic appearance of the ovaries, and ovulatory dysfunction. PCOS women have an increased prevalence of cardiovascular risk factors, such as increased blood pressure, insulin resistance (IR), and obesity. We have previously demonstrated that glucagon-like peptide-1 receptor agonists (GLP1RA) improve IR and obesity in a rat model of PCOS. Recent evidence suggests that Black Women (BW) with PCOS have an increased incidence of cardiometabolic risk factors compared to their White Women (WW) counterparts. We tested the hypothesis that GLP1RA has a differential effect on cardiometabolic risk factors in BW with PCOS versus WW with PCOS. Methods: The University of Mississippi Medical Center de-identified patient data from January 2013 – September 2022 were obtained from electronic medical records (EMR) using UMMC's Research Data Warehouse. Patients with 1) PCOS, 2) type 2 diabetes mellitus (T2DM), 3) documented hemoglobin A1c (HbA1c), and 4) GLP1RA prescription were identified. Cardiometabolic risk factors such as HbA1c, body mass index (BMI), and systolic and diastolic blood pressure (SBP and DBP respectively) were analyzed and compared between BW and WW with PCOS/T2DM. ICD-10 code of PCOS and T2DM: E28.2 and E11.9 respectively. Data are presented as mean ± standard deviation. Results: EMRs of 3756 PCOS patients were obtained. 13.8% of PCOS patients also had a diagnosis of T2DM with documented HbA1c, and 10.4% of them (54 patients) were on a GLP1RA. Of the 54 patients, 35 were BW. In our cohort, BW with PCOS/T2DM had a similar age (38 ± 12 vs 34 ± 11 years of age, p=NS), perimenopausal status, and smoking status as WW with PCOS/T2DM. Compared to WW with PCOS/T2DM on GLP1RA, BW women with PCOS/T2DM had significantly higher HbA1c (8.0 ± 1.0 vs 7.0 ± 0.7 %, p<0.05). There was no difference in BMI, SBP, or DBP between groups. Furthermore, on GLP1RA, there was no difference in the magnitude of change of HbA1c, BMI, SBP, or DBP in BW compared in WW on GLP1RA. Conclusions: BW with PCOS/T2DM had higher HbA1c on GLP1RA compared to WW counterparts. However, there was similar BMI, SBP, and DBP between the two groups. Furthermore, there was no difference in the magnitude of change in cardiometabolic parameters on GLP1RA between the two groups. Race/ethnicity may significantly effect HbA1c, but it does not appear to affect the efficaciousness of GLP1RA on the cardiometabolic profile in women with PCOS. NIGMS P20GM121334 and P20GM104357, NIDDK R21DK11350 and F30DK127527, NHLBI P01HL51971 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

LINC00092 derived from follicular fluid alleviated the symptoms of PCOS through inactivation of phosphatase and tensin homolog by recruiting KDM5A

Mounting literatures suggest that follicular fluid-derived exosomes (FF-Evs) influence the progression of progression of polycystic ovary syndrome (PCOS). The present study was designed to dissect the underlying mechanisms by which FF-Evs affect the PCOS. A rat model of PCOS was established using Letrozole induction. After treatment with FF-Evs, rats were examined for alterations in hormones, blood glucose, and lipid levels in serum, oestrus cycle, pathology in the ovaries, and apoptosis of ovarian cells. The functional rescue assays were performed to analyze the impact of long non-coding RNA 00092 (LINC00092) on PCOS rats. The cis-regulatory elements involved in the regulation of phosphatase and tensin homolog (PTEN) expression were analyzed using bioinformatic analysis, followed by verification of the mechanism. FF-Evs treatment ameliorated Letrozole-induced enhancement of weight, insulin resistance, dyslipidemia, and LH/FSH ratio, reduction of luteal cells, granulosa cells, and healthy follicles, prolonged oestrus, oestrous cycle arrest, ovarian tissue fibrosis, and ovarian cell apoptosis in rats, which were counteracted by treatment with shRNA targeting LINC00092. Regarding the mechanism, FF-Evs augmented LINC00092 expression in rats. LINC00092 bound to lysine demethylase 5 A (KDM5A), and KDM5A facilitated the demethylation of H3K4me3 to restrain the transcriptional activity of PTEN. Taken together, FF-Evs delivered LINC00092 repressed the transcriptional activity of PTEN by binding to KDM5A to enhance demethylation of H3K4me3, thereby reducing apoptosis in ovarian cells and alleviating PCOS symptoms.

Reduced expression of Il10, Stat3, Hoxa10, and Itgb3 in the embryo implantation site of rat model with prenatal androgen-induced polycystic ovary syndrome.

Impaired implantation due to the reduced endometrial receptivity considers an etiology for infertility in polycystic ovary syndrome (PCOS). In this context, we aimed to compare the expression of interleukin 10 (Il10), homeobox A10 (Hoxa10), signal transducer and activator of transcription 3 (Stat3), and β3-integrin (Itgb3) in the embryo implantation site of a prenatally-androgenized rat model of PCOS before and during gestation. PCOS rat model was created by the injection of testosterone prenatally. The uterine tissues were collected before pregnancy (day 0) and on days 0.5, 4.5, 5.5, and 8.5 of gestation in the PCOS rat model and controls (n=6; each group). RNA was extracted from the uterine samples and reverse transcribed to cDNA. Expression levels of Il10, Stat3, Hoxa10, and Itgb3 were measured using SYBR Green real-time RT-PCR and compared between the two groups. PCOS rats showed decreased expression levels of the Il10 on day 8.5 compared to control rats. The mRNA levels of Hoxa10, Itgb3, and Stat3 were significantly decreased in the PCOS group on day 0 as well as on days 0.5, 4.5, 5.5, and 8.5 for Hoxa10, Itgb3, and Stat3. The decreased gene expression of Il10, Hoxa10, Stat3, and Itgb3 in the PCOS rat model indicates the importance of the Il10 signaling axis as one of the possible disrupted mechanisms of endometrial receptivity in PCOS.

1,25-Dihydroxyvitamin D inhibits hepatic diacyglycerol accumulation and ameliorates metabolic dysfunction in polycystic ovary syndrome rat models.

Introduction: We aimed to evaluate the influence of 1,25-dihydroxyvitamin D (1,25(OH)2D) on metabolic dysfunction and elucidate its underlying mechanism using a rat model of polycystic ovary syndrome (PCOS). Methods: Twenty-four Sprague-Dawley rats were randomly divided into four groups: control group (CON, 2ml/kg of oral 0.5% CMC), 1,25VD group (oral 0.5% CMC and 2.5ug/kg intraperitoneal 1,25(OH)2D), PCOS group (1mg/kg oral letrozole), PCOS+1,25VD group (1mg/kg oral letrozole orally 2.5ug/kg intraperitoneal 1,25(OH)2D). The treatments were administered for 8weeks. Body weight, estrus cycle, insulin tolerance, and oral glucose tolerance of the rats in the different groups were assessed. The rats were euthanized at the 8thweeks, and plasma, ovarian, and liver samples were collected and analyzed. The hepatic lipid profile was characterized using HPLC/MRM. Results: Letrozole-induced PCOS rats exhibited increased weight, insulin resistance, postprandial glucose abnormalities, and dyslipidemia. Compared with the PCOS group rats, the PCOS+1,25VD group rats showed reduced body weight, increased sensitivity to insulin, decreased postprandial glucose, and elevated levels of high-density lipoprotein cholesterol. Moreover, abnormally increased liver concentrations of total diacylglycerol (DG) and DG species in the PCOS rats were reversed by treatment with 1,25(OH)2D. Additionally, hepatic DG and insulin sensitivity were correlated. Conclusion: 1,25(OH)2D inhibited hepatic DG accumulation and ameliorated metabolic dysfunction in PCOS rat models.

Protective effect of wuzibushen recipe on follicular development via regulating androgen receptor in polycystic ovary syndrome model rats

Objectives The study aimed to explore the role and mechanism of WZBS recipe on PCOS. Methods PCOS model was established. After modeling, PCOS rats were intragastrically administered with Diane-35 or WZBS recipe (6.93 g/kg/d). Then, the ovarian and uterine morphology were observed, the estrous cycle was assessed. HE and oil red O staining were conducted for ovarian morphological analysis and counting ovarian follicle and corpora lutea number. Furthermore, the serum content of testosterone (T) and sex-hormone-binding globulin (SHBG) were assessed by ELISA kits. The androgen receptor (AR), CX43 mRNA and protein expression were measured by q-PCR and Western blot. Results WZBS recipe increased uterine implanted blastocysts, reduced cystic dilated follicles, and normalized estrous cycle in PCOS rats. Meanwhile, WZBS recipe alleviated ovarian injury, increased mature follicles and corpora lutea number in PCOS rats. Moreover, WZBS recipe decreased serum T content, AR expression and increased serum SHBG content, CX43 expression in PCOS rats. Conclusions This study reveals that WZBS recipe may attenuate PCOS by protecting follicular development via down-regulating AR.

Ameliorative effect of sesame oil on experimentally induced polycystic ovary syndrome: A cross-link between XBP-1/PPAR-1, regulatory proteins for lipogenesis/steroids.

Polycystic ovary syndrome (PCOS) is a mixed endocrine/metabolic/reproductive disorder in women of reproductive age. Sesame oil (SO) contains sesame lignans & vitamin E with broad-spectrum antioxidant and anti-inflammatory effects. This study investigates the ameliorative effect of SO on experimentally induced PCOS and elucidates the possible molecular mechanisms with a deeper focus on the different signaling pathways involved. The study was carried out on 28 nonpregnant female Wister albino rats that were divided into four equal groups; Group I (control group) received oral 0.5% wt/vol carboxymethyl cellulose daily. Group II (SO group): orally administered SO (2 mL/kg body wt./day) for 21 days. Group III (PCOS group) received letrozole daily, 1 mg/kg, for 21 days. Group IV (PCOS + SO group): concomitantly administered letrozole and SO for 21 days. The serum hormonal and metabolic panel and the homogenate ATF-1, StAR, MAPK, PKA, and PI3K levels of the ovarian tissue were calorimetrically evaluated. However, endoplasmic reticulum (ER) stress was evaluated by ovarian XBP1 and PPAR-γ messenger RNA expression level using the qRT-PCR technique. Ovarian COX-2 was detected immunohistochemically. The results suggest that SO-treated PCOS rats showed a significantly improved hormonal, metabolic panel, inflammatory, and ER stress status with concomitant decreases in ATF-1, StAR, MAPK, PKA, and PI3K in ovarian rats compared to the correspondent values in PCOS without treatment. CONCLUSIONS: The protective effects of SO against PCOS are triggered by ameliorating regulatory proteins of ER stress, lipogenesis, and steroidogenesis through the PI3K/PKA and MAPK/ERK2 signaling cascades. SIGNIFICANCE STATEMENT: Polycystic ovary syndrome (PCOS) is the most common mixed endocrine-metabolic dysfunction among women within the reproductive period, with an estimated prevalence of 5%-26% worldwide. Doctors traditionally recommend metformin for PCOS patients. However, metformin is known to be associated with significant adverse effects and contraindications. This work aimed at shedding light on the ameliorative effect of sesame oil (SO), natural polyunsaturated fatty acids-rich oil, on the induced PCOS model. SO proved to have a marvelous effect on the metabolic and endocrine derangements in the PCOS rat model. We hoped to provide a valuable alternative treatment for PCOS patients to avoid the side effects of metformin and to help PCOS patients for whom metformin is contraindicated.

The ameliorative effect of CangFu Daotan Decoction on polycystic ovary syndrome of rodent model is associated with m6A methylation and Wnt/β-catenin pathway

Objective: This study investigates the effects of CangFu Daotan Decoction (CDD) on m6A methylation and Wnt/β-catenin pathway in rats with polycystic ovary syndrome (PCOS).Methods: The PCOS rat model was established by letrozole gavage. The rats were fed high-fat chow, and their body weight and blood glucose were recorded. The expressions of follicle-stimulating hormone(FSH), luteinizing hormone(LH), and testosterone(T) were quantified by ELISA. Chemical components in CDD were analyzed using UPLC-Q/TOF-MS. Based on network pharmacology methods, related targets of CDD on PCOS were screened. An enrichment analysis according to Tokyo Encyclopedia of Genes and Genomes (KEGG) was conducted to predict the potential signaling pathway of CDD in PCOS. The expressions of Wnt-1, β-Catenin, GSK-3β, C-MYC, Beclin1, LC3II, Bax, and PCNA were detected by western blotting. The expressions of Mettl3, Mettl14, Fto, Alkbh5, Ythdf1, and Ythdf2 were monitored by RT-PCR. The expressions of Mettl3, Fto, and Ythdf1 were detected by western blotting.Results: Letrozole and a high-fat diet induced ovarian dysfunction in rats, which was attenuated by CDD. CDD decreased blood glucose, LH, and T concentrations and increased FSH expression in PCOS. After removing duplicates, a total of 71 compounds were identified by UHPLC-Q/TOF-MS, among which terpenoids and flavonoids account for the main proportion. The clustering analysis showed that the active site of CDD might be in the Wnt/β-catenin pathway. CDD decreased the expressions of Wnt-1, β-Catenin, GSK-3β, C-MYC, Beclin1, LC3II, and Bax and increased PCNA expression in the ovarian tissue of PCOS rats. CDD decreased the m6A gene expressions of Mettl3, Mettl14, Fto, Alkbh5, Ythdf1, and Ythdf2 in peripheral blood and ovarian tissue of PCOS rats. CDD reduced the m6A proteins expressions of Mettl3, Fto, and Ythdf1 in the ovarian tissue of PCOS rats.Conclusion: CDD can regulate m6A modification and inhibit the Wnt/β-catenin signaling pathway in PCOS rats, thereby reducing body weight, lowering blood glucose levels, improving sex hormone disorders, and decreasing autophagy and apoptosis in ovarian tissue to promote the recovery of ovarian morphology.

MicroRNA-1298-5p in granulosa cells facilitates cell autophagy in polycystic ovary syndrome by suppressing glutathione-disulfide reductase.

The aim of this study was to investigate the effect and mechanism of action of miR-1298-5p in polycystic ovary syndrome (PCOS). Granulosa cells were isolated from follicular fluid of patients with PCOS and healthy women, and the expression of miR-1298-5p and glutathione-disulfide reductase (GSR) mRNA in these cells was evaluated using reverse transcription-quantitative polymerase chain reaction (qRT-PCR). Clinical data were obtained from all subjects, and reproductive hormones and endocrine indices were assayed to analyze the correlation between miR-1298-5p and clinicopathological characteristics of patients with PCOS. Following transfection with the miR-1298-5p mimic or inhibitor and/or pcDNA3.1-GSR, LC3 immunofluorescence and transmission electron microscopy were used to evaluate autophagy in the COV434 human granulosa cell line. Additionally, western blotting was performed to detect LC3-II, Beclin 1, and p62 protein levels in COV434 cells. The interaction between miR-1298-5p and GSR was also examined. A PCOS rat model was established and injected with the miR-1298-5p antagomir, followed by measurement of body and ovary weights, histological examination, and autophagosome observation. The protein expression levels of GSR, LC3-II, Beclin 1, and p62 were determined in rat ovaries. miR-1298-5p was expressed at a high level, and GSR was downregulated in granulosa cells from patients with PCOS. In COV434 cells, miR-1298-5p inversely mediated GSR expression, and miR-1298-5p mimic transfection promoted autophagy, whereas GSR overexpression blocked miR-1298-5p mimic-promoted autophagy. In PCOS rats, miR-1298-5p inhibition reduced autophagy and alleviated abnormalities in follicular development. Overall, miR-1298-5p enhances autophagy in granulosa cells by downregulating GSR, thereby affecting PCOS development.

Effect of berberine on serum hormonal levels and endometrial gene and protein signaling in a rat model of polycystic ovary syndrome

Purpose: To study the effect of berberine on serum hormonal levels, endometrial gene, and protein signaling in a rat model of polycystic ovary syndrome (PCOS).Methods: Eighty-seven healthy female Sprague Dawley (SD) rats were assigned to control group, PCOS model group, and berberine group, each with 29 rats. The rats were assigned to three groups: control, model, and berberine groups, each with 29 rats. A rat model of PCOS was established via the administration of letrozole (1 mg/kg) to rats in the model group continuously for 21 days through gavage. Rats in berberine group were also given letrozole at the same dose for the same duration through gavage, while rats in control group received physiological saline in place of berberine. Estrus cycle, serum hormonal levels, ovarian tissue morphology, estrogen receptor α (ERα) levels, and mRNA expression levels of protein kinase B subtype 3 (Akt-3β) were evaluated. RAS-C3 botulinum toxin substrate 1 (Rac1), human chromosome 10 deleted phosphatase (PTEN) and mouse sarcoma-like virus oncogene (KRAS) were also determined using appropriate procedures.Results: Berberine treatment resulted in a significant decrease in the mean body weight of PCOS rats, relative to model rats (p < 0.05). Estrus interval in model rats was significantly prolonged. Levels of testosterone and estradiol, and mRNA expression levels of ERα, Akt3, Racl, PTEN, and KRAS were significantly increased, while progesterone level was significantly decreased, relative to control (p < 0.05). Estrus interval in the berberine group was shortened and estrus cycle gradually returned to normal, while the levels of testosterone and estradiol, and mRNA levels of ERα were gradually restored, relative to untreated PCOS rats (p < 0.05). Moreover, mRNA levels of Akt3, RAC1, PTEN, and KRAS were significantly decreased, while progesterone level was significantly increased (p < 0.05).Conclusion: Berberine appreciably restores heat cycle and ovary morphology in PCOS rats, and regulates serum hormonal levels, and endometrium-related gene and protein signaling.

Elevated Insulin-Like Growth Factor-1-Induced Female Rats Perpetuate the Polycystic Ovary Syndrome Phenotype: Pathological Mechanism of Insulin-Like Growth Factor-1 in Polycystic Ovary Syndrome

Objectives: This study was designated to establish a polycystic ovary syndrome (PCOS) rat model with recombinant human insulin-like growth factor-1 (RH-IGF-1). We made assessment on the characteristics of hyperinsulinemia and hyperandrogenism in the rat model. Design: This study performed the characteristics of PCOS upon RH-IGF-1 injection and evaluated the disease process of PCOS syndrome caused by the insulin-resistant pathological condition of IGF-1 based on the comparative study of in vivo test. Setting: The experiment was conducted in the experimental research center of Yinzhou NO.2 hospital, Ningbo, Zhejiang Province, China. Materials and Methods: Thirty-four female Sprague Dawley immature rats aged 21 days were randomly divided into two groups. Those treated with RH-IGF-1 2 mg/100 g daily were in RH-IGF-1 group (n = 20), and those with 0.9% sodium chloride 0.2 mL/100 g daily were in the saline group (n = 14). The experiment was carried out in two stages. In stage I, rats were anesthetized upon the first estrous cycle in the saline group with tissue and blood samples collected (n = 7), and rats in the RH-IGF-1-treated group were anesthetized on the 5th day after vaginal opening (VO) (n = 10). In stage II, rats in the saline group were anesthetized after three complete cycles (n = 7), meanwhile, while on the 15th day after VO (n = 10) for those in the RH-IGF-1 group. Results: We have found that compared with the control group, rats injected with RH-IGF-1 expressed an early VO, disordered estrous cycle, polycystic ovaries, and significantly increased ovarian weight/body weight ratio. And from the perspective of hormone secretion, their androgen increased significantly and the insulin resistance index also elevated distinctly, possessing main characteristics similar to PCOS. Limitations: In this study, we were limited by the inability to examine IGF-1 in hypothalamus. IGF-1 in hypothalamus and in vitro experiments would be taken into consideration for further study in the future. Conclusions: These findings suggest that IGF-1 may be a key factor in the pathogenesis of PCOS, and the increase of androgen may be the pathological result, not the cause of PCOS.

Kisspeptin regulates the proliferation and apoptosis of ovary granulosa cells in polycystic ovary syndrome by modulating the PI3K/AKT/ERK signalling pathway

BackgroundThe development of polycystic ovary syndrome (PCOS) is closely correlated with apoptosis and oxidative stress in ovarian granulosa cells. Kisspeptin plays an important role in reproductive organ function. This study aimed to explore the role of kisspeptin in PCOS and oxidative stress-triggered apoptosis of ovarian granular cells.MethodsA PCOS rat model was established by injecting dehydroepiandrosterone (DHEA) and feeding the rats a high-fat diet. The RNA and protein levels of kisspeptin were analysed by quantitative PCR, western blotting, and histological staining. Tissue damage was evaluated using haematoxylin and eosin (H&E) staining. The viability and proliferation of human granulosa cell KGN were measured using the cell counting kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assays. Cell cycle and apoptosis were analysed by flow cytometry. Oxidative stress was analysed by measuring reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) levels.ResultsKisspeptin was downregulated in the ovarian granulosa cells of PCOS rats compared to those of control rats. Kisspeptin overexpression enhanced KGN cell proliferation and inhibited apoptosis. ROS generation was suppressed by kisspeptin, along with decreased levels of MDA and increased levels of the antioxidants GSH, SOD, and CAT. Kisspeptin activates PI3K/AKT and ERK signalling, and inactivation of ERK1/2 suppresses the protective role of kisspeptin in ovarian granulosa cells.ConclusionKisspeptin improves proliferation and alleviates apoptosis and oxidative stress in ovarian granulosa cells by activating PI3K/AKT and ERK signalling.

Evaluation of renal disturbance in animal models of polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is an endocrine disease characterized by hyperandrogenism and hyperinsulinemia, followed by luteinizing hormone and follicle-stimulating hormone deficiency. PCOS conditions cause metabolic disorders that increase uric acid levels and malondialdehyde (MDA) levels. Animal models of PCOS have been used extensively in research to study the pathogenesis, clinical characteristics, and treatment of PCOS. This study aimed to identify the pathological mechanisms underlying renal dysfunction in PCOS by observing several parameters, including blood urea nitrogen (BUN), creatinine, uric acid, and renal MDA levels. This research was an experimentally designed study using a Wistar rat (Rattus norvegicus) as an animal model of PCOS which were divided into three groups: negative control group (n = 6), Testosterone propionate (TP) induction group (n = 6), and estradiol valerate (EV) induction group (n = 6). According to statistical analysis it indicated that induction of TP and EV can increase blood uric acid levels in PCOS model rats (p < 0.05), TP induction can increase kidney BUN and MDA levels significantly (p < 0.05), However, the observation of creatinine levels did not show significant differences in all treatment groups (p > 0.05). Based on the results of this study, it can be concluded that the induction of animal models with TP can trigger significant renal damage compared to EV.