Effect of long non-coding RNA00963 on lipopolysaccharide-induced apoptosis of rat renal tubular epithelial cells via microRNA-150-5p/interleukin-1 receptor-associated kinase 1 axis
10.3969/j.issn.1006-6187.2024.12.012
- VernacularTitle:长链非编码RNA00963通过微小RNA-150-5p/白介素1受体关联激酶1轴对脂多糖诱导的大鼠肾小管上皮细胞凋亡影响的研究
- Author:
Wenxuan CUI
1
;
Yan WANG
;
Yang YU
;
Juan JIA
;
Haige ZHAO
Author Information
1. 071000保定,河北大学附属医院
- Publication Type:Journal Article
- Keywords:
Long-chain non-coding RNA00963;
MicroRNA-150-5p/interleukin-lreceptor-associated kinase 1 axis;
Lipopolysaccharide;
Renal tubular epithelial cells;
Apoptosis
- From:
Chinese Journal of Diabetes
2024;32(12):941-950
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of long-chain non-coding RNA00963 (LINC00963) on lipopolysaccharide (LPS) induced apoptosis in rat renal tubular epithelial cells via microRNA-150-5p (miR-150-5p)/interleukin-1 receptor-associated kinase 1 (IRAK1) axis. Methods Rat renal tubular epithelial cells NRK-52E were cultured in vitro. NRK-52E cells were divided into control group (Con),LPS group,LPS+si-NC group,LPS+si-LINC00963 group,LPS+si-LINC00963+inhibitor NC group,LPS+si-LINC00963+miR-150-5p inhibitor group,LPS+si-LINC00963+oe-NC group,and LPS+si-LINC00963+oe-IRAK1 group. MTT and EdU methods were used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis rate. ELISA kits were used to detect TNF-α,IL-6,and IL-10 levels,and commercial reagent kits were used to analyze malondialdehyde (MDA),superoxide dismutase (SOD),glutathione peroxide dismutase (GSH-Px) levels. The qRT-PCR was used to detect the expression levels of LINC00963,miR-150-5p,and IRAK1 genes in each group. Western blot was used to detect the expression of cysteine aspartic acid specific protease-3(Caspase-3),Bax,Bcl-2,and IRAK1 proteins in cells. Dual luciferase reporter gene and RNA pull down experiment were used to verify the relationship between miR-150-5p and LINC00963 and IRAK1. Results Compare with the Con group,the expression of LINC00963 and IRAK1 mRNA,the levels of TNF-α,IL-6,the content of MDA,apoptosis rate,the expression of Caspase-3,Bax,and IRAK1 proteins in NRK-52E cells were obviously increased (P<0.05),the expression of miR-150-5p,A490 (24 hours,48 hours) values,proliferation rate,the level of IL-10,the activities of SOD and GSH-Px,and the expression of Bcl-2 protein were obviously reduced (P<0.05) in the LPS group and LPS+si-NC group. Compare with the LPS+si-NC group,the expression of LINC00963 and IRAK1 mRNA,the levels of TNF-α,IL-6,the content of MDA,apoptosis rate,the expression of Caspase-3,Bax,and IRAK1 proteins in NRK-52E cells were obviously reduced (P<0.05),the expression of miR-150-5p,A490 (24 hours,48 hours) values,proliferation rate,the level of IL-10,the activities of SOD and GSH-Px,and the expression of Bcl-2 protein were obviously increased (P<0.05) in the LPS+si-LINC00963 group. Down-regulation of miR-150-5p expression or overexpression of IRAK1 could reduce the improvement effect of interference LINC00963 on LPS induced NRK-52E cell damage (P<0.05). Conclusions Interference with LINC00963 can regulate the miR-150-5p/IRAK1 axis to alleviate LPS-induced oxidative stress,inflammatory response,and apoptosis in NRK-52E cells.
