Circ_0114428 Regulates Sepsis-Induced Kidney Injury by Targeting the miR-495-3p/CRBN Axis.

Abstract

Septic acute kidney injury (AKI) is considered as a severe and common complication of sepsis, with complex pathogenesis. Recently, Circular RNA (circRNA) is considered to be implicated in this disease. This study was intended to elucidate the role of circ_0114428 and the potential mechanism of action in sepsis-induced kidney injury. Sepsis-induced kidney injury cell model was established in human kidney 2 (HK2) cells by the treatment of lipopolysaccharide (LPS). The expression of circ_0114428, CRBN mRNA, and miR-495-3p was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was assessed by cell counting kit-8 (CCK-8) assay. The inflammatory response was monitored according to the release of proinflammatory factors by enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was evaluated by flow cytometry assay. The activities of oxidative indicators were examined using the corresponding kits. Endoplasmic reticulum (ER) stress-related proteins and CRBN protein were quantified by western blot. RNA immunoprecipitation (RIP) assay was performed to ensure whether circ_0114428 could interact with Argonaute 2 (Ago2) protein. The potential miRNAs targeted by circ_0114428 were predicted by the bioinformatics tool and screened by RNA pull-down assay. The interaction between miR-495-3p and circ_0114428 or CRBN was validated by dual-luciferase reporter assay. The results showed that circ_0114428 and CRBN were upregulated in septic AKI serum specimens and LPS-induced HK2 cells. Circ_0114428 knockdown attenuated LPS-induced apoptosis, inflammation, oxidative stress, and ER stress, which were rescued by CRBN overexpression. Further analysis revealed that miR-495-3p was targeted by circ_0114428 and directly bound to CRBN, and circ_0114428 regulated CRBN expression by sponging miR-495-3p. Besides, miR-495-3p inhibition also reversed the effects of circ_0114428 knockdown. In conclusion, circ_00114428 knockdown attenuated LPS-induced HK2 cell injury by regulating CRBN expression via targeting miR-495-3p.