LncRNA NEAT1 accelerates renal fibrosis progression via targeting miR-31 and modulating RhoA/ROCK signal pathway

• Published in: American Journal of Physiology: Cell Physiology 2023-02, Vol.324 (2), p.C292-C306
• Main Authors: Chen, Yan, Huang, Chong, Duan, Zhi-Bin, Chen, Yan-Xia, Xu, Cheng-Yun
• Format: Article
• Language: English
• Subjects: Actin; Apoptosis; Connective tissue growth factor; Connective tissues; End-stage renal disease; Enzyme-linked immunosorbent assay; Fibrosis; Fluorescence in situ hybridization; Humans; Immunohistochemistry; Immunoprecipitation; In Situ Hybridization, Fluorescence; Inflammation; Kidney Diseases; MicroRNAs - genetics; MicroRNAs - metabolism; Non-coding RNA; Oxidation; Renal failure; Rho-associated kinase; rho-Associated Kinases - genetics; rho-Associated Kinases - metabolism; rhoA GTP-Binding Protein - genetics; rhoA GTP-Binding Protein - metabolism; RhoA protein; RNA, Long Noncoding - genetics; RNA, Long Noncoding - metabolism; Signal Transduction; Smooth muscle
• ISSN: 0363-6143; 1522-1563
• DOI: 10.1152/ajpcell.00382.2021

Renal fibrosis is the final pathway for chronic kidney disease to end-stage renal failure. Noncoding RNAs have been reported to play a crucial role in renal fibrosis. Here, the effects of long noncoding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) and miR-31 on renal fibrosis and their regulatory mechanism were evaluated. RT-qPCR was used to assess NEAT1, miR-31, and RhoA levels. Western blot was performed to analyze the expression of fibrosis markers, RhoA, rho-related kinase (ROCK1), and connective tissue growth factor (CTGF). RNA immunoprecipitation (RIP), fluorescence in situ hybridization (FISH), and luciferase reporter assays verified the interaction between miR-31 and NEAT1 or RhoA. Renal fibrosis and injury were observed by Masson and hematoxylin and eosin (H&E) staining. The expression level of inflammatory cytokines was detected by ELISA. Immunohistochemistry (IHC) was performed to examine the expression levels of α-smooth muscle actin (α-SMA) and RhoA in renal tissues. We showed that NEAT1 was highly expressed, whereas miR-31 was decreased in renal fibrosis. NEAT1 was found to directly bind miR-31 to positively regulate RhoA expression. Furthermore, NEAT1 silencing inhibited renal fibrosis and inflammation and suppressed the RhoA/ROCK1 signaling pathway. However, knockdown of miR-31 could reverse these effects. NEAT1 silencing or overexpression of miR-31 alleviated renal fibrosis in vivo. In conclusion, NEAT1 accelerates renal fibrosis progression via negative regulation of miR-31 and the activation of RhoA/ROCK1 pathway, thereby upregulating the expression level of CTGF, providing a theoretical basis for treatment and prognostic evaluation of renal fibrosis.