Effect of LncRNA CRNDE on LPS-induced alveolar epithelial cell injury by regulating the miR-338-3p/KLF6 axis
Objective To investigate the effect of LncRNA CRNDE on LPS-induced alveolar epithelial cell injury by regulating the miR-338-3p/KLF6 axis.Methods Human alveolar epithelial cells A549 were separated into CK group,LPS group,LPS+si-NC group,LPS+si-CRNDE group,LPS+si-CRNDE+inhibitor NC group,LPS+si-CRNDE+miR-338-3p inhibitor group,LPS+si-CRNDE+oe-NC group,and LPS+si-CRNDE+oe-KLF6 group;qRT-PCR was applied to detect the expression levels of CRNDE,miR-338-3p,and KLF6 of cells in each group;MTT method was applied to detect cell proliferation;flow cytometry was applied to detect cell apoptosis rate;ELISA kit was applied to detect TNF-α,IL-10,and IL-1β levels;commercialized reagent kit was applied to analyze MDA,GSH-Px,and SOD levels;Western blot was applied to detect the expression levels of KLF6,cleaved-caspase-3,Bax,and Bel-2 proteins in cells.Dual luciferase reporter gene experiment was applied to verify the relationship between miR-338-3p and RNDE and KLF6.Results Compared with the CK group,the expression of CRNDE and KLF6 mRNA,the levels of TNF-α,IL-1β,content of MDA,apoptosis rate,the expression of caspase-3,Bax,and KLF6 proteins in A549 cells in the LPS group and LPS+si-NC group were obviously increased,the expression of miR-338-3p,cell viability,the level of IL-10,activities of SOD and GSH-Px,and expression of Bcl-2 protein were obviously reduced(P＜0.05);compared with the LPS+si-NC group,the expression of CRNDE and KLF6 mRNA,the levels of TNF-α,IL-1β,content of MDA,apoptosis rate,the expression of caspase-3,Bax,and KLF6 proteins in A549 cells in the LPS+si-CRNDE group were obviously reduced,the expression of miR-338-3p,cell viability,the level of IL-10,activities of SOD and GSH-Px,and expression of Bcl-2 protein were obviously increased(P＜0.05);interference with miR-338-3p expression or overexpression of KLF6 was able to reduce the down-regulation of CRNDE expression and improve LPS-induced A549 cell damage(P＜0.05).Conclusion Down-regulation of CRNDE can regulate the miR-338-3p/KLF6 axis to alleviate LPS-induced damage to A549 cells.
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