The role of hepatitis B virus surface protein in inducing Sertoli cell ferroptosis

Abstract

AbstractBackgroundsHepatitis B virus infection could result in male infertility with sperm defects and dysfunction. Sertoli cells are essential for testis function and play a crucial role in spermatogenesis. Sertoli cell death contributes to spermatogenesis impairment, leading to poor sperm quality. Ferroptosis has been implicated as a mechanism of Sertoli cell death. The issue in studying the relationship between hepatitis B virus and Sertoli cell ferroptosis has not yet been addressed.ObjectivesTo explore the mechanisms underlying ferroptosis in hepatitis B virus‐exposed Sertoli cells.Materials and methodsHuman Sertoli cells were treated in vitro with levels of 25, 50, and 100 μg/mL of hepatitis B virus surface protein (HBs). Cell viability and levels of glutathione, malondialdehyde, cellular ferrous ion (Fe2+), lipid peroxidation, and N6‐methyladenosine in Sertoli cells were detected. The level of glutathione peroxidase 4, transferrin receptor 1, ferritin heavy chain, tripartite motif (TRIM) 37, methyltransferase like 3, and insulin‐like growth factor 2 mRNA binding protein 2 was examined. Cell transfection was carried out to alter expression of ferroptosis‐related proteins. qPCR and immunoblotting were performed to measure protein expression level. Immunoprecipitation was applied to determine the protein and protein‐RNA interaction. Luminescence analysis was performed to identify the target of methyltransferase like 3.ResultsHBs exposure triggered ferroptosis featured with increased intracellular Fe2+ ion, reduced cell viability and expression of glutathione peroxidase 4 in Sertoli cells. HBs treatment significantly increased TRIM37 expression, which suppressed glutathione peroxidase 4 expression through ubiquitination. TRIM37 silencing attenuated the effect of HBs exposure‐regulated cell viability and ferroptosis. HBs upregulated N6‐methyladenosine modification in TRIM37 3′‐UTR by increasing methyltransferase like 3 expression. The binding of N6‐methyladenosine reader insulin‐like growth factor 2 mRNA binding protein 2 and TRIM37 3′‐UTR enhanced the stability of TRIM37 mRNA.ConclusionHBs can decrease human Sertoli cell viability by promoting ferroptosis induced by the loss of glutathione peroxidase 4 activity through TRIM37‐mediated ubiquitination of glutathione peroxidase 4. The findings highlight the role of TRIM37/glutathione peroxidase 4 signaling responsible for ferroptosis regulation in hepatitis B virus‐infected Sertoli cells.