The Effect Of Overexpression Of The Enhancer Of Zeste Homolog 1 (Ezh1) Gene On Aristolochic Acid-Induced Injury In Hk-2 Human Kidney Proximal Tubule Cells In Vitro

Background: Acute kidney injury (AKI) involves the renal tubular epithelium. The enhancer of zeste homolog 1 (EZH1) gene has a role in cell development and differentiation. This study aimed to investigate the effect of overexpression of the EZH1 gene on aristolochic acid-induced injury in HK-2 human kidney proximal tubule epithelial cells in vitro.Material/Methods: The HK-2 cells were cultured and treated with aristolochic acid and the effects of aristolochic acid-injury were evaluated using a cell counting kit-8 (CCK-8) assay. Overexpression of EZH1 used gene plasmid transfection into HK-2 cells. The cell apoptosis rate and levels of intracellular reactive oxygen species (ROS) were measured using flow cytometry. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were performed to determine the expressions of inflammatory cytokines including interleukin (IL)-1 beta, IL-6, tumor necrosis factor-alpha (TNF-alpha), apoptosis-related genes, and the downstream target genes of NF-kappa B signaling pathway, including NFKBIA, CXCL8, and cyclin D1.Results: Aristolochic acid inhibited HK-2 cell viability, induced cell apoptosis, increased the levels of ROS and inflammatory cytokines, including IL-1 beta, IL-6, TNF-alpha, and activated the NF-kappa B pathway. Overexpression the EZH1 gene inhibited HK-2 cell apoptosis, reduced ROS levels, and down-regulated the expressions of IL-1 beta, IL-6, TNF-alpha, Bax and Cyt C mRNA and protein, and increased the expressions of Bcl-2 and NFKBIA, CXCL8 and cyclin D1, indicating that overexpression of EZH1 suppressed NF-kappa B signaling in aristolochic acid-injured HK-2 cells.Conclusions: Overexpression of EZH1 reduced HK-2 cell injury induced by aristolochic acid in vitro by inhibition of NF-kappa B signaling.