Hyperin Alleviates Triptolide-Induced Ovarian Granulosa Cell Injury by Regulating AKT/TSC1/mTORC1 Signaling

Abstract

Premature ovarian insufficiency (POI) is characterized by the loss of ovarian function before 40 years of age and affects approximately 1% of women worldwide. Caragana sinica is a traditional Miao (a Chinese ethnic minority) medicine that improves ovarian function and follicular development. In the present study, we aimed to investigate the effect of active ingredients of C. sinica on POI and determine underlying mechanisms. Herein, the chemical composition of the C. sinica compound was analyzed using ultra-high-performance liquid chromatography, which identified hyperin (HR) as one of the main ingredients in C. sinica. Then, interaction targets of HR and POI were predicted and analyzed using network pharmacology and bioinformatics. The effect of HR on triptolide (TP)-induced granulosa cell injury was evaluated, and the underlying mechanism was explored based on bioinformatic results. A total of 100 interaction targets for POI and HR were obtained. The protein-protein interaction network of identified interaction targets emphasized the topological importance of AKT1. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that HR might regulate POI by modulating the mechanistic target of rapamycin (mTOR) signaling pathway. In addition, the KEGG graph of the mTOR signaling pathway revealed that AKT phosphorylation inhibits the TSC1/2, while TSC1/2 activation inhibits the expression of mTORC1. The fundamental experiment revealed that HR increased proliferation, progesterone receptor levels, and estradiol levels decreased by TP in KGN cells. Additionally, HR alleviated TP-induced apoptosis and G1/G1 phase arrest in KGN cells. Western blotting demonstrated that HR increased the phosphorylation of AKT and mTORC1 and decreased TSC1 expression in TP-induced KGN cells. Collectively, our findings revealed that HR alleviates TP-induced granulosa cell injury by regulating AKT/TSC1/mTORC1 signaling, providing insight into the treatment of POI.

The effects of natural products and bioactive ingredients of traditional Chinese medicine on apoptosis of ovarian granulosa cells

“…The mechanisms underlying the protective effects of hyperoside, anthocyanin, isorhamnetin, quercetin, genistein, kurarinone, and baicalein against apoptosis of GCs are related to antioxidant pathways (Li et al, 2022; Li, Chen, et al, 2021; Luo et al, 2020; Rashidi et al, 2019; Tao et al, 2017; Wang, Fan, et al, 2019; Xiang et al, 2017). In addition, the flavonoids hyperoside, astragalin, quercetin, isorhamnetin, and genistein are phytoestrogens reported to improve hormone production and protect against apoptosis of GCs induced by triptolide, CdCl 2 , and H 2 O 2 (Li, Chen, et al, 2021; Luo et al, 2020; Nna et al, 2017; Wei et al, 2016; You et al, 2021). Scutellarin significantly alleviated zearalenone‐induced cell damage, restored cell cycle arrest, and reduced apoptosis and positive signal expression of cleaved caspase‐3 and Bax/Bcl‐2 in vitro and in vivo (Yi et al, 2021).…”

XinJiaCongRongTuSiZiWan protects triptolide-induced rats from oxidative stress injury via mitophagy mediated PINK1/Parkin signaling pathway

“…However, plenty of studies have exhibited severe reproductive system toxicity in TP-treated animals and patients 5 , 6 . TP can significantly induce ovarian damage, resulting in follicular atresia, a decrease in the number of corpus luteum and in the gonad index, and the destruction of ovarian microstructure 7 , 8 . Oxidative stress is one of the core molecular mechanisms involved in the toxicity of TP, and has been demonstrated to be tightly associated with follicular atresia and ovulation disorders, including premature ovarian failure (POF) and polycystic ovary syndrome (PCOS) 9 , 10 .…”