Naringenin and morin reduces insulin resistance and endometrial hyperplasia in the rat model of polycystic ovarian syndrome through enhancement of inflammation and autophagic apoptosis

Abstract

Polycystic Ovary Syndrome (PCOS) is a gynecologic disorder with unsatisfactory treatment options. Hyperandrogenism and insulin resistance (IR) are two symptoms of PCOS. The majority of PCOS patients (approximately 50% to 70%) have IR and moderate diffuse inflammation of varying degrees. We investigated in-vitro and in-vivo effects of naringenin, morin and their combination on PCOS induced endometrial hyperplasia by interfering with the mTORC1 and mTORC2 signaling pathways. The vaginal smear test ensured the regular oestrous cycles in female rats. Serum cytokines (TNF-α & IL-6) were assessed using the ELISA test, followed by in-vivo and in-vitro determination of prominent gene expressions (mTORC1& C2, p62, LC3-II, and Caspase-3 involved in the inflammatory signaling mechanisms through RT-PCR, western bloting, or immunohistochemical analysis. In addition, the viability of naringenin or morin treated cells was determined using flow cytometry analysis. The abnormal oestrous cycle and vaginal keratosis indicated that PCOS was induced successfully. The recovery rate of the oestrous cycle with treatments was increased significantly (P<0.01) when compared to the PCOS model. Narigenin, morin, or a combination of the two drugs substantially decreased serum insulin, TNF-α, IL-6 levels with improved total anti-oxidant capacity and SOD levels (P<0.01). Treatments showed suppression of HEC-1-A cells proliferation with increased apoptosis (P<0.01) by the upregulation of Caspase-3 expression, followed by downregulation of mTORC, mTORC1, and p62 (P<0.01) expressions with improved LC3-II expressions (P<0.05) respectively. The histological findings showed a substantial increase in the thickness of granulose layers with improved corpora lutea and declined the number of cysts. Our findings noticed improved inflammatory and oxidative microenvironment of ovarian tissues in PCOS treated rats involving the autophagic and apoptotic mechanisms demonstrating synergistic in-vitro and in-vivo therapeutic effects of treatments on PCOS-induced endometrial hyperplasia.

Therapeutic Effects of Silibinin Against Polycystic Ovary Syndrome Induced by Letrozole in Rats via Its Potential Anti-Inflammatory and Anti-Oxidant Activities

“…The enhancement of ovarian histomorphological characteristics might be attributed to an anti-androgenic mechanism 54 of the silibinin as evident by the decrease in testosterone hormone level demonstrated in this study and the alleviation of endometrial hyperplasia might be attributed to the improved structural and functional microenvironment of the ovarian and uterine tissues. 55 , 56 The incomplete restoration of ovarian and uterine tissues in rats treated by silibinin alone is probably attributed to insufficient time-duration (19 days) of the treatment. Further investigations are needed to investigate the long-term treatment of silibinin with different doses and route of administration.…”

Naringenin improves ovarian health by reducing the serum androgen and eliminating follicular cysts in letrozole‐induced polycystic ovary syndrome in the Sprague Dawley rats

“…Similar results were seen, possibly due to the low duration of naringenin treatment and without any other combination, Daine-35, as used earlier (Wu, Yang, Han, et al 2022). However, naringenin treatment did not show normalization of the estrous cycle within the given duration of treatment as reported in the earlier studies (Hong et al, 2019;Yang et al, 2022). Add-on therapy of naringenin with metformin has been F I G U R E 1 1 Analysis of Estrous cycle/ovarian cycle analysis, body and organ weights, and oral glucose tolerance test in response to combined therapy of naringenin and metformin.…”

“…The dose of naringenin has been used earlier as 20 mg/kg body weight (b.wt.) for the determination of gut microbiota (Wu, Yang, Han, et al 2022), steroidogenic enzymes and antioxidants (Hong et al, 2019), and endometrial hyperplasia in the PCOS ovary (Yang et al, 2022). Further, in our research based on 4‐hydroxyisolucine (4‐HIL) PCOS management, we found 4‐HIL MED 50 mg/kg body weight in a similar PCOS model system (Indian patent reference no.…”

“…Twenty-five (42-day-old) female rats were randomly assigned into five groups of 5-6 rats each: the control (sham), letrozole (PCOS) (PHR-1540-1G, Sigma-Aldrich Chemicals Private Limited, Bangalore, India), metformin + letrozole wt.) for the determination of gut microbiota (Wu, Yang, Han, et al 2022), steroidogenic enzymes and antioxidants (Hong et al, 2019), and endometrial hyperplasia in the PCOS ovary (Yang et al, 2022).…”

Advancements in lead therapeutic phytochemicals polycystic ovary syndrome: A review

“…Furthermore, Yang et al disclosed that naringin combined with morin could inhibit the proliferation of human endometrial adenocarcinoma cells. In their study, naringin inhibited PCOS-induced endometrial hyperplasia and regulated autophagy by blocking the mammalian target of rapamycin complex 1 (mTORC1)/mammalian target of rapamycin complex 2 (mTORC2) signaling, and morin increased the levels of Caspase-3 and LC3-phosphatidylethanolamine conjugate (LC3-II) ( Yang et al, 2022 ). In addition, naringin could change the elevated serum insulin level and maintain the typical morphology of the ovary in rats with PCOS ( Wu et al, 2022 ).…”