Letrozole-induced Polycystic Ovary Syndrome Research Articles

Metabolomics combined with network pharmacology to explore the mechanisms of modified Guishen pill to ameliorate polycystic ovary syndrome

BackgroundThe modified Guishen pill (MGP) has a prominent therapeutic effect on polycystic ovary syndrome (PCOS). However, its mechanism is still unclear. This study aimed to uncover the mechanism of MGP for PCOS treatment through a comprehensive strategy integrating metabolomics and network pharmacology. MethodsA letrozole-induced PCOS model was used to evaluate ovarian function in rats. Plasma metabolomics was used to authenticate differential metabolites and enriched related pathways using the MetaboAnalyst platform. Network pharmacology was utilized to explore the endogenous targets of MGP treatment for PCOS. Finally, the potential targets and related biological functions were verified experimentally. ResultsMGP improved PCOS symptoms by regulating abnormal levels of sex hormones and alleviating ovarian pathological changes in rats; fifty-four potential differential metabolites involved in MGP treatment for PCOS, and the hub genes derived from network pharmacology were consistent with the metabolomic analysis results to varying degrees. The comprehensive analysis identified that a key novel target for endothelial nitric oxide synthase (eNOS/NOS3), five key metabolites (ornithine, citrulline, l-glutamic acid, acetylornithine, and hydroxyproline), and one pathway (arginine and proline metabolism) were related to the therapy of PCOS with MGP. Subsequently, we verified the localization and expression of eNOS in the ovaries, and it significantly improved insulin resistance, apoptosis, and oxidative stress in letrozole-induced PCOS rats. ConclusionOur work reveals the complex mechanism of MGP therapy for PCOS. This study is a successful paradigm for elucidating the pharmacological mechanism of the traditional Chinese medicine compound.

Selected Toxicity Study on Ethanolic Extract of Ocimum gratissimum (Linn.) Leaves in Letrozole-Induced Polycystic Ovarian Syndrome in Wistar Rats

Polycystic Ovarian Syndrome (PCOS) is a metabolic endocrine disorder that is common in women of reproductive age. Several synthetic drugs such as metformin are treatment options of PCOS but the side effects associated with the use of metformin have continued to limit their acceptability; Thus, necessitate the investigation of the safety of Ocimum gratissimum leaves in vivo. The toxicity of ethanolic extract of Ocimum gratissimum leaf (EEOGL) at 50 and 100mg/kg body weight dosage on PCOS was investigated in female Wistar rats. Twenty female Wistar rats with an average weight of 170.81±5.25g were allocated into 5 groups (A-E) of four animals each: group A animals (control) received distilled water while the letrozole-induced (1mg/kg body weight (B.Wt)) rats- groups B, C, D, and We were administered distilled water, co-administration of 7.14mg/kg B.Wt of metformin and 2mg/kg B.Wt of clomiphene citrate, 50mg/kg and 100mg/kg of EEOGL for 21 days respectively. The liver and kidney function indices were determined after each rat was sacrificed and blood collected by jugular puncturing. The data collected were subjected to analysis of variance and Duncan multiple range test with statistical significance set at p<0.05. The result reveals a significant increase (p<0.05) and a significant decrease (p<0.05) at doses of 50mg/kg and 100mg/kg of EEOGL on kidney function indices as well as liver function indices respectively. Therefore, the administration of EEOGL at 50 and 100 mg/kg B.W. to letrozole-induced PCOS rats shows mild alterations in the assayed toxicological indices.

Naringenin regulates gut microbiota and SIRT1/ PGC-1ɑ signaling pathway in rats with letrozole-induced polycystic ovary syndrome

PurposeTo evaluate the effect of naringenin on improving PCOS and explore the mechanism. MethodsFirstly, we carried out differential gene expression analysis from transcriptome sequencing data of human oocyte to screen the KEGG pathway, then the PCOS-like rat model was induced by letrozole. They were randomly divided into four groups: Normal group (N), PCOS group (P), Diane-35 group (D), and Naringenin group (Nar). The changes of estrus cycle, body weight, ovarian function, serum hormone levels, glucose metabolism, along with the expression of SIRT1, PGC-1ɑ, claudin-1 and occludin of the ovary and colon were investigated. Furthermore, the composition of the gut microbiome of fecal was tested. ResultsBy searching the KEGG pathway in target genes, we found that at least 15 KEGG pathways are significantly enriched in the ovarian function, such as AMPK signaling pathway, insulin secretion, and ovarian steroidogenesis. Interestingly, naringenin supplementation significantly reduced body weight, ameliorated hormone levels, improved insulin resistance, and mitigated pathological changes in ovarian tissue, up-regulated the expression of PGC-1ɑ, SIRT1, occludin and claudin-1 in colon. In addition, we also found that the abundance of Prevotella and Gemella was down-regulated, while the abundance of Butyricimonas, Lachnospira, Parabacteroides, Butyricicoccus, Streptococcus, Coprococcus was up-regulated. ConclusionOur data suggest that naringenin exerts a treatment PCOS effect, which may be related to the modulation of the gut microbiota and SIRT1/PGC-1ɑ signaling pathway. Our research may provide a new perspective for the treatment of PCOS and related diseases.

The Role of a High-Fat, High-Fructose Diet on Letrozole-Induced Polycystic Ovarian Syndrome in Prepubertal Mice.

This study aims to investigate the effects of a high-fat, high-fructose (HF/HFr) diet on metabolic/endocrine dysregulations associated with letrozole (LET)-induced Polycystic Ovarian Syndrome (PCOS) in prepubertal female mice. Thirty-two prepubertal C57BL/6 mice were randomly divided into four groups of eight and implanted with LET or a placebo, with simultaneous administration of an HF/HFr/standard diet for five weeks. After sacrifice, the liver and blood were collected for selected biochemical analyses. The ovaries were taken for histopathological examination. The LET+HF/HFr group gained significantly more weight than the LET-treated mice. Both the LET+HF/HFr and the placebo-treated mice on the HF/HFr diet developed polycystic ovaries. Moreover the LET+HF/HFr group had significantly elevated testosterone levels, worsened lipid profile and indices of insulin sensitivity. In turn, the HF/HFr diet alone led to similar changes in the LET-treated group, except for the indices of insulin sensitivity. Hepatic steatosis also occurred in both HF/HFr groups. The LET-treated group did not develop endocrine or metabolic abnormalities, but polycystic ovaries were seen. Since the HF/HFr diet can cause substantial metabolic and reproductive dysregulation in both LET-treated and placebo mice, food items rich in simple sugar—particularly fructose—and saturated fat, which have the potential to lead to PCOS progression, should be eliminated from the diet of young females.

Effects of Kelulut Honey on Oestrus Cycle Regulation and Histomorphological Changes in Letrozole-Induced Polycystic Ovary Syndrome Rats: A Preliminary Study.

Polycystic ovary syndrome (PCOS) is a complex reproductive, metabolic, and endocrine disorder that affects women of reproductive age. Kelulut honey is stingless bee honey that possesses anti-inflammatory, anti-cancer, anti-diabetic, and potent antioxidative activities in most conditions. However, its value in improving PCOS remains to be elucidated. Thus, this preliminary study aimed to determine the effective dose of Kelulut honey in oestrus cycle regulation and ovarian histomorphological changes in letrozole-induced PCOS rats. PCOS was induced in all-female Sprague Dawley (SD) rats with 1 mg/kg/day of letrozole except for the control group for 21 days. Kelulut honey was then orally administered to the PCOS rats at the dose of 0.5, 1, or 2 g/kg/day, respectively, for 35 days. The oestrous cycle was determined through vaginal smears, while ovarian histomorphological changes were observed by haematoxylin and eosin (H&E) staining. The untreated PCOS rats were characterised by irregular oestrous cyclicity, hyperglycaemia, and aberrant ovarian histology. In this study, Kelulut honey (1 g/kg/day) increased the number of corpus luteum and antral follicles (p < 0.05), improved the cystic follicle, and normalised the oestrus cycle (p < 0.05). This preliminary study demonstrated that Kelulut honey, particularly at a dose of 1 g/kg/day, has the potential to alleviate oestrus cycle dysregulation and ovarian histomorphological changes occurring in PCOS.

Histomorphometric and biochemical activities of bioactive component of Cyperus esculentus tubers extract on letrozole-induced polycystic ovarian syndrome and cholesterol homeostasis in female Sprague-Dawley rats

Polycystic ovary syndrome (PCOS) is a heterogeneous endocrine and metabolic disorder, causes irregular menstrual cycles, dyslipidemia, excessive body weight, oxidative stress, hyperandrogenism, and infertility. This study focused on the impact of Cyperus esculentus (CES) on letrozole-induced PCOS in female Sprague-Dawley rats. Twenty (20) normal rats and twenty (20) PCOS rats (150–200 g, 8 weeks old) were randomly divided into four groups of ten ( n = 10) rats each. Group A served as normal control group received 2 mL of normal saline, group B treated with 500 mg/kg body weight of CES , group C PCOS control group received 2 mL of normal saline, and group D PCOS rats post-treated with 500 mg/kg body weight of CES daily through gastric gavage for 30 days. Estrus cyclicity, body and ovaries weights, biochemical and histological parameters were measured. Observed irregular estrus cyclicity and multiple cysts in PCOS rats, increase glycemia, ovarian weight, triglycerides, total cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, malondialdehyde, luteinizing hormone (LH), testosterone, and decrease high-density lipoprotein cholesterol, estradiol, progesterone, catalase, reduced glutathione (GSH), and superoxide dismutase levels, compared with control. The intervention of CES ameliorated and restored the estrus cyclicity reproductive hormone, biochemical, and structural alterations. Moreover, CES significantly decreased cystic follicles, LH, and testosterone levels, but increased estradiol concentration. This plant may be beneficial in the management and treatment of PCOS-related reproductive and metabolic disorders.

Liquiritin ameliorates metabolic and endocrine alterations in a mouse model of polycystic ovary syndrome

Abstract Objective: Altered bile acid transformation induces low-grade chronic inflammation and may play an important role in the pathophysiology of polycystic ovary syndrome (PCOS). Liquiritincan regulate bile acid metabolism and anti-inflammatory properties; however, limited information is available regarding its therapeutic potential in PCOS. Methods: Female C57BL/6 mice were randomly assigned into four groups (n = 6 mice/group): the control, letrozole or dehydroepiandrosterone-induced PCOS groups, PCOS + 20 mg/kg liquiritin group, and control + liquiritin groups. After 21 days of treatment, the mice were euthanized, and the associated metabolism indications were investigated. Ovarian histological examinations were performed, and serum hormone concentration was measured. The expression of key genes involved in steroid hormone synthesis, ovarian follicle development, and ovulation was assessed. Results: Liquiritin reduced fasting blood glucose levels and increased insulin sensitivity compared to the PCOS group. Liquiritin also significantly decreased serum levels of total testosterone (P &lt; 0.001) and dehydroepiandrosterone sulfate (P &lt; 0.05) in the PCOS group. Histomorphological inspection of ovaries from the liquiritin group revealed fewer cystic dilated follicles than in the PCOS group. Moreover, liquiritinsignificantly (P &lt; 0.01) decreased Cyp17a1, Cyp19a1, Fshr, Hsd3b2, Runx2, and Ccn2 mRNA expression compared to letrozole-induced PCOS. Conclusion: Liquiritin may be safe and helpful in ameliorating PCOS-associated hyperandrogenemia and hyperglycemia. However, clinical trials investigating different liquiritin dosages are needed to confirm these findings.

Effects of Fagonia indica on Letrozole-Induced Polycystic Ovarian Syndrome (PCOS) in Young Adult Female Rats.

Polycystic ovarian syndrome is a multidisciplinary endocrinopathy of reproductive-aged women that provokes insulin resistance, hyperandrogenism, cardiovascular problems, obesity, and menstrual complications. The present study was designed to investigate the effectiveness of ethanolic extract of Fagonia indica in letrozole-induced PCOS young adult female rats. HPLC was carried out to find the phenolic and flavonoid content of the ethanolic extract of Fagonia indica. Twenty-five female rats were taken and initially divided into two groups: group I (control group) and group II (PCOS group). PCOS was induced by letrozole given orally by gavage. Body weight was recorded weekly and vaginal cytology was analyzed daily. After induction of disease, the PCOS group is further divided into four groups (n = 5): group II (positive control with PCOS), group III (metformin 20 mg/kg treated group), group IV (ethanolic extract of Fagonia indica 500 mg/kg treated group), and group V (metformin plus Fagonia extract). At the end of experimental period, the blood sample of each rat was collected and serum was separated by centrifugation. Afterwards hormonal analysis, lipid profile and liver functioning tests were performed. Ovaries were removed and preserved for histopathological findings while the liver of each rat was stored for the determination of antioxidant potential assessment. Fagonia indica was found to possess quercetin as one of the major flavonoid phytoconstituents. The plant extract exhibited its beneficial effects by restoring hormonal balance, lipid profile, and liver functioning markers. Treatment with F. indica reduced body weight, resolved ovarian cysts, and showed positive effects on follicular growth. Treatment with plant also increased the levels of antioxidant enzymes. This study validates the potential of Fagonia indica for the amelioration of metabolic, as well as, hormonal disturbances that occurred in PCOS.

Evaluation of Formulated Herbal Syrup (Containing Fennel, Anise, and Celery) on the Letrozole-Induced Polycystic Ovary Syndrome Model

Background: Polycystic ovary syndrome (PCOS) is a complex endocrine disorder associated with irregular menstrual cycles, hyperandrogenism, obesity, and reduced fertility. Objectives: The present study aimed to formulate herbal syrup based on Iranian traditional medicine (ITM) and evaluate its effect on the letrozole-induced PCOS model in female rats. Methods: The herbal syrup contains anise, fennel, and celery seed extracts. Five different formulations were made with different percentages of additive components. Quality control and stability tests were performed on the selected formulation. During the in vivo step, 6 groups of rats were evaluated: The control group (received carboxymethyl cellulose 1% as a vehicle) and the other 5 groups (received letrozole 1 mg/kg orally for 21 days). During 21 days, daily vaginal smears were examined to detect irregularities of the estrous cycle. After induction of PCOS, rats were orally administered with herbal syrup (1, 2, 4 mL/kg) or metformin (200 mg/kg) for 28 days. Moreover, body and ovarian weights, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, progesterone, and testosterone were measured. Finally, ovarian tissues were isolated for histological examination. Results: The best formulation of the syrup contained the plant extract (totally 10%), sugar (50%), sodium benzoate (0.1%), and potassium sorbate (0.1%). Body weight was significantly increased in all groups compared to the control group, and after treatment, a significant weight reduction was seen in the metformin and 1-mL/kg dose group. Following PCOS induction, ovary weight was significantly increased, while after treatment, it showed a significant decrease. After 21 days of letrozole administration, induction of PCOS was confirmed by the irregularities in estrous cycles and an increase in testosterone and LH levels. After treatments with the syrup, LH levels significantly decreased in all groups (P &lt; 0.05), and serum testosterone and FSH levels significantly decreased in the 2-mL/kg dose group compared to the PCOS group (P &lt; 0.05). Estradiol and progesterone levels significantly increased in the treatment groups in a dose-dependent manner. Histological studies of metformin and herbal syrup groups exhibited normal follicular development with fewer and smaller cystic follicles compared to the PCOS group. Conclusions: The herbal syrup made from anise, fennel, and celery seed extracts improved serum levels of sex hormones, recovered the ovarian morphology in PCOS-induced rats, and can be a good candidate for further clinical trials.

Effects of N-acetylcysteine and metformin treatment on the stereopathological characteristics of uterus and ovary.

In this study, the stereo-pathological effect of metformin and N-acetyl cysteine is evaluated on the uterus and ovary of polycystic ovary syndrome (PCOS) mice. 96 mature females (8-weekold, weight of 20–30 gr) BALB/c mice were classified into 6 groups including the control group (n= 16), letrozole-induced PCOS group (n=16), PCOS + metformin (n=16), PCOS+NAC (n=16) and a separate control group for NAC (n=16). Another PCOS group was maintained for a month to make sure that features remain till the end of the study. Testosterone level, vaginal cytology and stereological evaluations were assessed. Vaginal cytology in letrozole-receiving mice showed a diestrus phase continuity. Testosterone level, body weight, uterine weight, endometrial volume, myometrial volume, gland volume, stromal volume, epithelial volume, vessel volume, daughter and conglomerate glands, endometrial thickness, and myometrial thickness exhibited an increasing trend in the uterus of PCOS mice. While normal gland and vessel length decreased in the PCOS group. Ovarian volume, corticomedullary volume, primary follicles, secondary follicles, and ovarian cysts were increased in PCOS ovaries. While corpus luteum, primordial, graafian, and atretic follicles showed a decline in the PCOS group. NAC and metformin, however, managed to restore the condition to normal. Given the prevalence of PCOS and its impact on fertility, the use of noninvasive methods is of crucial significance. NAC can control and treat pathological parameters and help as a harmless drug in the treatment of women with PCOS.

Metformin treatment ameliorates endocrine-metabolic disturbances in letrozole-induced PCOS mice model by modulating adiponectin status

In reproductive aged women, polycystic ovarian syndrome (PCOS) is a complex endocrine condition. It's generally linked to metabolic diseases (MDs), thereby increasing the risk of infertility, cardio metabolic events, and other comorbidities in PCOS women. Adiponectin is a circulating protein made by adipocytes that has been shown to have an inverse relationship with MDs. Metformin, an insulin-sensitizing biguanide, is the world's most widely prescribed treatment for type 2 diabetes and PCOS. The effects of Metformin and the involvement of adiponectin in endocrine-metabolic abnormalities in experimentally induced PCOS mice were examined in this study. Letrozole (6 mg/kg) was administered per orally (p.o) for a period of 21 days for the induction of PCOS, followed by dose of Metformin (150 mg/kg, p.o) for 60 days using 0.9% saline water as vehicle. The findings revealed that LET-treated mice experienced PCOS-like symptoms, including increased Serum LH, decreased Serum estrogen, progesterone and FSH levels, increased body weight, altered ovarian cytoarchitecture, with multiple cysts. In addition, there was an increase in serum cholesterol, triglycerides, fasting blood glucose, plasma interleukin-6 ( IL6). In the ovary we observed a decrease in levels of superoxide dismutase (SOD), glutathione (GSH) and increase in lipid peroxidation (MDA). These changes were linked to lower levels of circulating adiponectin and were altered when the mice were given metformin. The findings revealed that metformin, via altering circulating adiponectin, appear to alleviate endocrine-metabolic abnormalities and inflammation-related comorbidities associated with LET-induced PCOS. • PCOS is a complex endocrine-metabolic illness. • Oxidative stress pathogenic component in PCOS. • PCOS is associated with higher levels of IL6. • Metformin increases circulating adiponectin a common point in PCOS. • Metformin Mitigates endocrine-metabolic problems associated with PCOS.

Suppression of PCSK9/NF-kB-dependent pathways by acetate ameliorates cardiac inflammation in a rat model of polycystic ovarian syndrome

AimEndocrinometabolic disorders in women of reproductive age, including polycystic ovarian syndrome (PCOS) has contributed to increased prevalence of cardiovascular disease (CVD) risk and its attendant complications. Acetate, the most abundant endogenously produced short chain fatty acid has been linked to metabolic health. However, the impact of acetate on CVD-driven pathologies in PCOS is unknown. The present study therefore investigated the effects of acetate on cardiometabolic abnormalities associated with PCOS in rat model, and the possible involvement of PCSK9/NF-kB-dependent pathways. Materials and methodsEight-week-old female Wistar rats were allotted into four groups (n = 6) and the groups received vehicle, acetate (200 mg/kg), letrozole (1 mg/kg) and letrozole plus acetate respectively. The administrations were done once daily by oral gavage and lasted for 21 days. Key findingsIn letrozole-induced PCOS rats characterized with insulin resistance, glucose dysregulation, elevated plasma testosterone and decreased 17-β estradiol as well as degenerated ovarian follicles, there was a significant increase in plasma and cardiac lipid/lipoproteins, lipid peroxidation, inflammatory mediators (NF-kB and TNF-α), γ-glutamyl transferase/lactate dehydrogenase and lactate content, PCSK9 and reduction in plasma and cardiac antioxidants (glutathione peroxidase and reduced glutathione) and plasma nitric oxide synthesis (eNOS and NO) compared with the control rats. In addition, immunohistochemical assessment of cardiac tissue showed severe expression of inflammasome in letrozole-induced PCOS rats compared with the control rats. Nevertheless, supplementation with acetate significantly attenuated these alterations. SignificanceThe present results suggest that acetate protects against cardiac inflammation in a rat model of PCOS by suppression of PCSK9 and NF-kB-dependent mechanisms.

Bio-enhancement of Soy Isoflavones (Genistein & Daidzein) Using Bacillus coagulans in Letrozole Induced Polycystic Ovarian Syndrome by Regulating Endocrine Hormones in Rats.

Soy isoflavone (SIF), a natural phytoestrogen, is used in the condition of hormonal imbalance. These isoflavones generally have low solubility resulting in low bioavailability and bioactivity. It is reported that trans-glycosylation by cyclodextrin glycosyltransferase (CGTase) is widely utilized for increasing the solubility and bioavailability of isoflavones. Present investigation was aimed to study the effect of Bacillus coagulans (a probiotic) in potentiating the bioactivity of soy isoflavones in letrozole-induced PCOS. Initial consideration was focused on proving CGTase assay of B. coagulans. After that, animal study was performed to check the enhancement of bioactivity of SIF along with B. coagulans. A total of 36 rats, separated into six groups (6 rats in each), were used. Group I received vehicles, group II received letrozole (1mg/kg) for 21days, and group III animals were administered with soy isoflavones (SIF-100mg/kg). In the case of group IV, V, and VI, animals received SIF (100mg/kg) along with B. coagulans 0.65, 3.25, and 6.50mg/kg, respectively. Treatment was given for 2weeks after induction of disease. All the animals were sacrificed at the end of the study and endpoint parameters were performed. Present investigation revealed that combination of SIF with B. coagulans showed hormone restoration, reduce oxidative stress, recovery in the menstrual cycle, and improvement in ovarian physiology. SIF (genistein & daidzein) together withB. coagulansexhibits a beneficial role in the enhancement of the bioactivity of soy isoflavones.Further, it showed that a higher dose ofB. coagulans(6.50mg/kg) is more effective in ameliorating the PCOS symptoms.

Diacerein ameliorates letrozole-induced polycystic ovarian syndrome in rats

Polycystic ovary syndrome (PCOS) is the most common gynaecological endocrine disease that causes anovulatory infertility. The current study aimed to explore the possible role of diacerein (DIA), an IL-1β inhibitor, in treating letrozole-induced PCOS in rats that exhibit the metabolic and endocrinal criteria of PCOS patients. PCOS was induced in female Wistar rats by the oral administration of letrozole (1 mg/kg, per orally, p.o.) for 21 days. Rats were then treated with DIA (25 mg/kg/day, p.o.), DIA (50 mg/kg/day, p.o.), or metformin (2 mg/100 g/day, p.o.) for 14 days after the PCOS induction. PCOS resulted in a significantly higher body weight, ovarian weight, ovarian size, and cysts, as well as an elevation in serum testosterone, LH, insulin, glycemia, and lipid profile levels. All of these effects were significantly reduced by the DIA administration. Additionally, DIA remarkably inhibited the letrozole-induced oxidative stress in the ovaries, muscles, and liver by reducing the upraised levels of malondialdehyde and total nitrite and increasing the suppressed levels of superoxide dismutase and catalase. DIA enhanced the protective proteins Keap-1, Nrf2, and OH-1 levels. Finally, DIA inhibited the elevated mRNA levels of NLRP3 and caspase-1, the up-regulated inflammatory cytokines IL-6, TNF-α, and the IL-1β/NFκB signaling pathway. Our results proved that DIA ameliorates letrozole-induced PCOS through its antioxidant and anti-inflammatory properties.

Partially purified non-polar phytocomponents from Aloe barbadensis Mill. gel restores metabolic and reproductive comorbidities in letrozole-induced polycystic ovary syndrome rodent model- an “in-vivo” study

Ethnopharmacological relevanceIn India, Kumaryasava, a popular Aloe barbadensis Mill. gel preparation has therapeutic value in treatment of female reproductive disorders like menstrual disturbances and menopausal problems. Despite their widespread use, only a limited number of studies have probed into the scientific evidence for their varied bioactivities. In this regard, studies have demonstrated that Aloe vera gel has the potential to modulate steroidogenic activity in letrozole induced polycystic ovary syndrome (PCOS) rat. However, isolation and identification of the bioactive molecule/s from Aloe vera gel and studying their molecular targets will underpin the treatment regime for PCOS. Material and methodsThe Partially Purified Non-Polar Phytocomponents (PPNPP)- LP1 and LP3 were isolated from the petroleum ether extract of Aloe vera gel by column chromatography. Based upon the GC-MS analysis, LP1 and LP3 comprised of n-Hexadecanoic acid and Campesterol acetate with an abundance of 97.07%, and 96.07% respectively. For evaluation of their bioactivities, eighty 3–4 months female Balb/c mice were classified as 10 groups with 8 animals in each group. Groups were control (C), PCOS (0.5 mg/kg/day Letrozole orally for 21days), PCOS treated orally for 60 days with Aloe vera gel (AVG) (10 mg/kg/day) (PCOS + AVG), PCOS treated orally for 60 days with petroleum ether extract (PE) of Aloe vera gel (25 μg/kg/day) (PCOS + PE), PCOS treated orally for 60 days with LP1 (0.5 μg/kg/day) (PCOS + LP1), PCOS treated orally for 60 days with commercially available pure compound-n-Hexadecanoic acid (HA) (0.5 μg/kg/day) (PCOS + HA), PCOS treated orally for 60 days with LP3 (0.01 μg/kg/day) (PCOS + LP3), PCOS treated orally for 60 days with commercially available pure compound- Campesterol acetate (CA) (0.01 μg/kg/day) (PCOS + CA), PCOS treated orally for 60 days with Metformin (100 mg/kg/day) (PCOS + Metformin) and PCOS treated orally for 60 days with DMSO (Vehicle) (PCOS + DMSO). Body weight, Oral glucose tolerance test, lipid profile, fasting glucose, insulin, estrus cycle, hormonal profile, gene expression of gonadotropin receptors (Fshr and Lhr), steroid receptors (Ar, Esr1, Esr2 and Pgr) and steroidogenic markers (Star, Hsd3b1, Cyp19a1 and Amh) were analysed in the ovaries. Polycystic ovarian morphology was assessed through histopathological changes of ovary. Toxicity markers- SGOT, SGPT and creatinine were also measured at the end of the study. ResultsMice treated with letrozole demonstrated significant increase in body weight, glucose intolerance, fasting insulin levels, HOMA-IR, triglycerides levels as well as testosterone levels, and a significant decline in the progesterone levels as compared to the control animals. PCOS animals also exhibited arrested estrus cyclicity, disrupted ovarian histopathology with the presence of multiple peripheral cysts and abnormal gene expression of gonadotropin receptor, steroid receptor and steroid markers. Oral administration of AVG, PE extract of AVG, LP3 and metformin greatly alleviated these complications in PCOS animals. ConclusionThe above findings indicate the effectiveness of LP3, isolated from Aloe vera gel against letrozole induced PCOS in mice and may be used in the treatment of PCOS as an alternative to metformin.

Histomorphological Changes in a Rat Model of Polycystic Ovary Syndrome and the Contribution of Stevia Leaf Extract in Modulating the Ovarian Fibrosis, VEGF, and TGF-β Immunoexpressions: Comparison with Metformin.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder of fertile females. It has been reported that stevia leaf extract (SLE) has antidiabetic and antihyperlipidemic properties. Therefore, the current study hypothesized and investigated the role and mechanistic aspects of a natural sweetener; SLE in treating a rat model of letrozole-induced PCOS and to compare it with metformin. Thirty-five female Wistar albino rats were divided into 5 groups: control, PCOS-induced group (letrozole, 1 mg/kg/d, for 21 days), SLE, metformin, and combination-treated groups (300 mg/kg/d, for the next 28 days in SLE and metformin-treated groups). Vaginal smears were done. The levels of glucose, lipid, and hormonal profiles were measured in the serum meanwhile, malonyl dialdehyde (MDA), superoxide dismutase (SOD), and tumour necrosis factor (TNF-α) were measured in the ovary. Ovarian sections were subjected to hematoxylin and eosin, Masson, and immunohistochemical identification of VEGF and TGF-β followed by morphometric analysis. PCOS rats showed altered hormonal and lipid profiles, in addition to hyperglycemia. Also, the ovarian tissue levels of MDA and TNF-α were elevated, and SOD was decreased. Numerous cystic follicles, decrease/absence of corpora lutea, interstitial fibrosis with positive VEGF and TGF-β immunoreactivity were evident. SLE improved all altered parameters. SLE showed potential therapeutic merits in letrozole-induced PCOS via anti-inflammatory, antioxidant, anti-fibrotic, and angiogenesis regulating mechanisms. Its effects were almost comparable to metformin, and the combination of both has no further synergistic effect.

Diacerein ameliorates induced polycystic ovary in female rats via modulation of inflammasome/caspase1/IL1β and Bax/Bcl2 pathways.

Polycystic ovary syndrome (PCOS) is a very common gynecological disease during childbearing period and markedly affects female fertility. Until now, there are no studies evaluating the possible curative effect of diacerein (DIA) in induced PCOS. For the first time, we aimed in current model to study the effect of DIA (50 mg/kg/day) orally for 3 weeks on experimentally induced PCOS by letrozole (1 mg/kg/day) for 3 weeks. We measured rats' body weight changes, levels of serum insulin, anti-Müllerian hormone (AMH), testosterone, inflammasome, caspase1, and total anti-oxidant capacity (TAC). Moreover, we measured ovarian tissue parameters as malondialdehyde (MDA), interleukin 1β (IL1β), real-time polymerase chain reaction (rt-PCR) of Bcl2-associated X protein (Bax), and interleukin 10 (IL10) gene expression changes. Furthermore, histopathological features and anti-apoptotic marker B cell lymphoma 2 (Bcl2) immunoexpression changes were evaluated. Our results showed that letrozole markedly induced PCOS as manifested by significant increase in serum testosterone, insulin, AMH, rats' body weights, ovarian tissue MDA, IL1β, inflammasome, and caspase1 but decrease of serum TAC. In addition, gene expression of Bax increased but IL10 gene expression decreased. Ovaries showed the typical histopathological changes of PCOS with no immunoexpression of Bcl2. DIA was greatly able to ameliorate letrozole-induced PCOS changes in rats mainly via prevention of IL1β, and improving metabolic disturbances, and its anti-apoptotic, anti-oxidant, and anti-inflammatory effects with further regulation of inflammasome/caspase1/IL1β and Bax/Bcl2 pathways.

Role of high-fat diet on letrozole-induced polycystic ovarian syndrome in rats

Polycystic ovary syndrome (PCOS) is one of the most prevalent hormonal disorders in women of reproductive age. Letrozole (LET)-induced PCOS is a good model but has drawbacks such as the absence of metabolic changes. Hence, in the present study, we aimed to develop a new animal model combining a high-fat diet (HFD) and LET.Female Wistar rats were divided into a control group, LET group, and LET + HFD (45% energy from fat) group.Compared with the control group, the LET and LET + HFD groups showed ovarian cysts and elevated testosterone levels, whereas oestradiol and progesterone levels were reduced. The LET + HFD group displayed significant changes in body weight, as well as in levels of triglycerides, fasting blood glucose, direct bilirubin, alanine aminotransferase, and uric acid; in terms of glucose intolerance and insulin resistance, the LET + HFD group showed better results than the LET group. Compared with the control group, elevated levels of tumour necrosis factor-α were detected in both the LET and LET + HFD groups.The addition of HFD to the LET model afforded good metabolic aberrations, along with ovarian cysts. In contrast, the LET-only model failed to demonstrate metabolic anomalies observed in the human PCOS condition.

Lactiplantibacillus plantarum CCFM1019 attenuate polycystic ovary syndrome through butyrate dependent gut-brain mechanism.

Polycystic ovary syndrome (PCOS) is an endocrine disorder that affects women of reproductive age. The gut microbiota has been shown to play a vital role in the pathogenesis of PCOS. Agents that target microbes in the gut may be promising therapeutic strategies for PCOS. Herein, a letrozole-induced PCOS model was used to test five Lactiplantibacillus plantarum strains for their ability to alleviate PCOS symptoms and their effect on the gut-brain axis. Lp. plantarum CCFM1019 attenuated the pathological changes in the ovaries and restored testosterone and luteinising hormone levels. However, metabolic disorders induced by letrozole treatment were not significantly reversed by these strains. Meanwhile, alteration of gut microbial diversity and enrichment of the short-chain fatty acid producers Lachnospira and Ruminococcus_2 were observed after Lp. plantarum CCFM1019 intervention. Compared with letrozole-treated rats, those treated with Lp. plantarum CCFM1019 exhibited higher butyrate and polypeptide YY levels, possibly due to the regulation of G protein-coupled receptor 41 expression. These results demonstrated that Lp. plantarum CCFM1019 attenuated letrozole-induced PCOS symptoms in rats. A butyrate-dependent gut-brain mechanism may be involved in this protective effect.

Gallic acid reverses ovarian disturbances in mice with letrozole-induced PCOS via modulating Adipo R1 expression

Research questionWomen are increasingly suffering from polycystic ovary syndrome (PCOS). Its pathophysiology is still unknown, though. The purpose of this study was to ascertain how gallic acid affected the pathophysiology of the ovary in an animal model of polycystic ovary syndrome. We also showed the potential mechanism of adiponectin involvement in endocrine metabolic changes in PCOS mice and the function of adiponectin, which appear to be frequent factors in PCOS. DesignEighteen adult female Parkes strain mice (Age: 4–5 weeks) having body weight of 16–21 g were separated into three groups at random with 6 animals in each group as follows: Group I serving the control, received water and normal diet for 81 days; group II received oral gavage administration of letrozole (LETZ)(6 mg/kg b.w.daily), which was dissolved in 0.9 % NaCl solution for 21 days for the induction of PCOS and left untreated for 60 days; Group III received oral gavage administration of LETZ (6 mg/kg) for first 21 days followed by the administration of gallic acid (GA) (75 mg/kg b.w. orally daily) for 60 days. ResultsWe found LETZ-treated mice experienced PCOS-like symptoms, including increased Serum testosterone, LH/FSH ratio, body and ovarian weight, blood glucose, serum insulin levels and inflammatory Cytokines. We also found decreased serum estrogen, oxidant capacity and enzyme activity and altered ovarian cytoarchitecture, with multiple cysts apart from irregular estrous cycle. Furthermore, mRNA expression levels CYP11a, CYP19a1, Kitl, PTGS2 and Adipo R1 were decreased. Furthermore, LETZ-induced PCOS mice when treated with GA we observed decrease in testosterone, LH, LH/FSH ratio, blood glucose, serum insulin and inflammatory cytokines. GA treatment in PCOS mice also increased estrogen levels, and oxidant capacity as well as enzyme activity. Furthermore mRNA expression levels of CYP11a1, CYP19a1, KITL, PTGS2 and Adipo R1 were also increased in LETZ+GA treated mice. These changes were linked to lower levels of circulating adiponectin and were altered when the mice were administered with gallic acid. ConclusionGallic acid might be a potential therapy in treating PCOS by regulating endocrine and metabolic abnormalities that are brought on by a drop in adiponectin levels along with hyperandrogenism. Additionally, adiponectin seems to be a frequent factor in PCOS. In addition to reducing inflammation-related comorbidities linked to LETZ-induced PCOS, GA enhances mRNA expression levels CYP11a, CYP19a1, Kitl and PTGS2 and hence reduces endocrine and metabolic abnormalities.