Genistein Improves Polycystic Ovary Syndrome by Regulating Oxidative Stress and Inflammation via Targeting Serum Amyloid A1

Abstract

Background: Polycystic ovary syndrome (PCOS) is a widespread complex endocrine and metabolic disease. Genistein has been found to play a vital role in multiple human diseases. This study aims to investigate the effects of genistein on the development of PCOS and further explore the underlying molecular mechanisms. Methods: A PCOS mouse model was established through treating C57BL/6 mice with dehydroepiandrosterone (DHEA). The mice were divided into Control group (n = 6), PCOS group (n = 6), PCOS+Genistein (5, 10, or 20 mg/kg; n = 6/each group) groups to evaluate the effects of genistein on PCOS development. Later, PCOS+Genistein+Ad-NC group (n = 6) and PCOS+Genistein+Adenovirus-serum amyloid A1 (Ad-SAA1) group (n = 6) were designed to explore the potential mechanism of genistein in PCOS. Tumor necrosis factor-α (TNF-α)-induced human granulosa-like tumor (KGN) cells were utilized to establish an in vitro PCOS cell model. In in vitro experiments, there were multiple groups, including Control group (n = 3), TNF-α group (n = 3), Genistein group (n = 3), TNF-α+si-NC (n = 3), TNF-α+si-SAA1 (n = 3), TNF-α+Genistein (n = 3), TNF-α+Genistein+pcDNA (n = 3), and TNF-α+Genistein+SAA1 (n = 3). The in vitro experiments were performed to investigate the effects of genistein on KGN cells and the underlying mechanisms. Hematoxylin and Eosin (HE) staining was applied to ovarian tissue for observing ovarian morphology in mice. The levels of luteinizing hormone (LH), estradiol (E2), follicle stimulating hormone (FSH), DHEA, testosterone, interleukin-1β (IL-1β) and interleukin-6 (IL-6), and glutathione-peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) were evaluated using the matched enzyme-linked immunosorbent assay (ELISA) kits, respectively. The relative level of (SAA1) in KGN cells and mouse ovarian tissues was measured by real-time quantitative polymerase chain reaction (RT-qPCR). Western blot was conducted to detect the protein expression of SAA1 and nuclear factor kappa B (NF-κB) signaling-related proteins. Results: DHEA reduced corpus luteum and elevated cystic follicles in PCOS ovarian tissues (p < 0.05). Genistein dramatically attenuated the pathological changes in ovarian tissues and influenced the serum levels of LH, FSH and E2 (p < 0.05). Moreover, SAA1 was upregulated in TNF-α-treated KGN cells and PCOS mouse tissues (p < 0.05). SAA1 knockdown alleviated TNF-α-induced oxidative stress and inflammation in KGN cells (p < 0.05). Genistein downregulated SAA1 expression in TNF-α-treated KGN cells (p < 0.05), and the improvement effects of genistein on oxidative stress and inflammation were blocked by SAA1 overexpression in TNF-α-treated KGN cells and DHEA-induced PCOS mice (p < 0.05). Conclusions: Our findings indicated that genistein targeted SAA1 to alleviate the development of PCOS by repressing the activation of the NF-κB signaling pathway.