Gandou Bushen Decoction Alleviates Ovarian Tissue Damage in TX Mice with Hepatolenticular Degener-ation by Inhibiting Ferroptosis and Endoplasmic Reticulum Stress
Objective To investigate the protective effect of Gandou Bushen Decoction(GDBSD)against ovarian damage in mice with hepatolenticular degeneration(HLD)and its molecular mechanism.Methods TX mice were used to establish a model of HLD,and then they were divided into HLD group,penicillamine group,and GDBSD group,while DL mice were established as normal control group.Body weight,ovarian weight,and ovarian coefficient were measured.The gonadotropin ovulation induction method was used to observe ovulation.HE staining was used to observe the histomorphology of mouse ovaries,and transmission electron microscopy was used to observe the ultrastructure of ovarian tissue.Colorimetry was used to measure serum iron content,the TBA method was used to measure the content of malondialdehyde(MDA)in ovarian tissue,and ELISA was used to measure the levels of reduced glutathione(GSH)and oxidized glutathione(GSSG)in ovarian tissue.Western blotting was used to measure the levels of the ferroptosis markers prostaglandin-endoperoxide synthase 2(PTGS2)and glutathione peroxidase 4(GPX4)in ovarian tissue,as well as the expression levels of endoplasmic reticulum stress pathway-related proteins,including glucose-regulated protein 78(GRP78),protein kinase RNA-like en-doplasmic reticulum kinase(PERK),phosphorylated PERK(p-PERK),eukaryotic initiation factor 2α(eIF2 α),phosphorylated eIF2α(p-eIF2α),activating transcription factor 4(ATF4),and C/EBP homologous protein(CHOP).Results HE staining showed that the HLD group had severe damage of oocyte morphology and structure and a significant increase in atretic follicles,and transmission electron microscopy showed obvious mitochondrial wrinkling and the manifestations of endoplasmic reticulum stress such as swelling and degran-ulation.Compared with the normal control group,the HLD group had significant reductions in ovarian weight(P＜0.05)and the num-ber of ovulations(P＜0.05),significant increases in serum iron and the levels of MDA and GSSG in ovarian tissue(P＜0.05),and sig-nificant reductions in GSH and GSH/GSSG(P＜0.05),as well as significant increases in the expression levels of PTGS2,GRP78,p-PERK,p-eIF2α,ATF4,and CHOP and a significant reduction in the expression level of GPX4 in ovarian tissue(P＜0.05).Compared with the HLD group,the GDBSD group had significant improvements in follicular morphology and the structure of mitochondria and en-doplasmic reticulum,a significant increase in the number of ovulations(P＜0.05),significant reductions in serum iron and the levels of MDA and GSSG in ovarian tissue(P＜0.05),and significant increases in GSH and GSH/GSSG in ovarian tissue(P＜0.05),as well as significant reductions in the expression levels of PTGS2,GRP78,p-PERK,p-eIF2α,and CHOP and a significant increase in the expres-sion level of GPX4 in ovarian tissue(P＜0.05).Conclusion GDBSD can alleviate ovarian damage induced by copper deposition in TX mice,possibly by inhibiting ferroptosis and endoplasmic reticulum stress mediated by the PERK pathway.
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