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

Abstract

BackgroundAcute 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/MethodsThe 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)-1β, IL-6, tumor necrosis factor-α (TNF-α), apoptosis-related genes, and the downstream target genes of NF-κB signaling pathway, including NFKBIA, CXCL8, and cyclin D1.ResultsAristolochic acid inhibited HK-2 cell viability, induced cell apoptosis, increased the levels of ROS and inflammatory cytokines, including IL-1β, IL-6, TNF-α, and activated the NF-κB pathway. Overexpression the EZH1 gene inhibited HK-2 cell apoptosis, reduced ROS levels, and down-regulated the expressions of IL-1β, IL-6, TNF-α, 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-κB signaling in aristolochic acid-injured HK-2 cells.ConclusionsOverexpression of EZH1 reduced HK-2 cell injury induced by aristolochic acid in vitro by inhibition of NF-κB signaling.

Aristolochic Acid-Induced Nephrotoxicity: Molecular Mechanisms and Potential Protective Approaches

“…Inflammation in a rat model of AA was also reported by Kholia et al [75]. and in HK-2 cells by Wang et al [65]. AA has been reported to increase the activity of caspases, resulting in apoptosis generation [4,66].…”

“…Apoptosis was also observed in renal tubular epithelial cells (NRK52E) treated with AA [63], in cultured LLC-PK1 [64], in proximal tubular epithelial cells (PTECs) by Pozdzik and colleagues [13], and in HK-2 cells by Wang et al [65]. AA has been reported to increase the activity of caspases, resulting in apoptosis generation [4,66].…”

Macrophages-Derived, LRG1-Enriched Extracellular Vesicles Exacerbate Aristolochic Acid Nephropathy Via A TGFβR1-Dependent Manner

“…damage through the production of pro-in ammatory mediators to generate the reactive oxygen species in AAN (Allard et al 2013, Wang et al 2019, Pozdzik et al 2010). In the current study of experimental AAN the prominent macrophage accumulation seen is consistent with studies of human AAN and previous studies of experimental AAN (Novitskaya et al 2014, Lu et al 2016.…”

Sinigrin attenuates angiotensin II‑induced kidney injury by inactivating nuclear factor‑κB and extracellular signal‑regulated kinase signaling in vivo and in vitro

“…Wu et al (11) indicated that NF-κB signaling plays an important role in Ang II-induced hypertensive renal damage. Additionally, previous studies have confirmed that the inhibition of NF-κB or ERK signaling can attenuate the progression of acute renal damage (10,12). Therefore, the NF-κB and ERK signaling pathways may be considered as targets for the treatment of hypertension-induced renal injury.…”