Study on the Involvement of GSH/GPX4 in Bisphenol A-induced Ferroptosis of Granulosa Cells in Small-tailed Han Sheep (Ovis aries)
ZHANG Yue1,2, WANG Xiang-Yu2, HE Yu2, ZHAO Sheng-Guo1,*, CHU Ming-Xing2,*
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Abstract：Bisphenol A (BPA), a common environmental endocrine disruptor, can induce apoptosis of cells, affect the function of ovarian granulosa cells, and pose a threat to the development of animal husbandry. This study aimed to preliminarily investigate the mechanism by which BPA induced ferroptosis in granulosa cells of Small-tailed Han sheep (Ovis aries) via the glutathione (GSH)/glutathione peroxidase 4 (GPX4) pathway. The viability of ovarian granulosa cells was assessed using the CCK-8 assay. The cells were treated with varying concentrations of BPA (0, 50, 100, 200, and 400 μmol/L) for 24, 48, and 72 h. Based on the results, the optimal conditions were determined to be a BPA concentration of 200 μmol/L and a treatment duration of 24 h. To validate the effect of BPA on ferroptosis in these cells, the experiment was divided into 3 groups: Control group, BPA-treated group (200 μmol/L BPA), and ferroptosis inhibition group (200 μmol/L BPA + 2 μmol/L Ferrostatin-1). To evaluate the key markers of ferroptosis, the expression of ferroptosis related genes and proteins were quantified by qRT-PCR and Western blot. Intracellular reactive oxygen species (ROS) levels were detected using the 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) method, while Fe2+, malondialdehyde (MDA), and glutathione (GSH) levels were quantified using commercial assay kits. The results showed that treatment with 200 μmol/L BPA extremely significantly increased the contents of Fe2+, ROS, and MDA (P<0.01), and extremely significantly decreased the level of GSH (P<0.01). mRNA level of solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), and ferritin heavy chain 1 (FTH1) was significantly upregulated (P<0.05), whereas GPX4 and SLC7A11 protein abundance significantly decreased (P<0.05) and FTH1 protein extremely significantly increased (P<0.01). Treatment with 2 μmol/L Ferrostatin-1 extremely significantly decreased Fe2+, ROS, and MDA contents (P<0.01), extremely significantly elevated GSH levels (P<0.01). The protein levels of GPX4 and SLC7A11 in the ferroptosis inhibition group were significantly higher than those in the BPA treatment group (P<0.05), while the FTH1 protein level was extremely significantly decreased (P<0.01). These results indicated that Ferrostatin-1 alleviated BPA-induced ferroptosis. This research demonstrated that BPA exposure triggered oxidative stress and disrupted iron metabolism in granulosa cells of Small Tail Han sheep by impairing the GSH/GPX4 axis, ultimately leading to ferroptosis. The addition of Ferrostatin-1 in vitro exhibited a significant antagonistic effect on this process. This study adopted a targeted ferroptosis strategy to mitigate the damage of environmental pollutants to sheep reproductive performance, providing a critical theoretical foundation for high fecundity and stress resistance breeding of sheep.
张越, 王翔宇, 何雨, 赵生国, 储明星. GSH/GPX4参与双酚A诱导小尾寒羊颗粒细胞铁死亡的研究[J]. 农业生物技术学报, 2026, 34(6): 1221-1231.
ZHANG Yue, WANG Xiang-Yu, HE Yu, ZHAO Sheng-Guo, CHU Ming-Xing. Study on the Involvement of GSH/GPX4 in Bisphenol A-induced Ferroptosis of Granulosa Cells in Small-tailed Han Sheep (Ovis aries). 农业生物技术学报, 2026, 34(6): 1221-1231.
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