TGF-β1 Changes The Phenotype Of Glomerular Mesangial Cells And Increases The Synthesis Of Extracellular Matrix Through LncRNA Uc.412

Objective:Transforming growth factor β1(TGF-β1),as an important member of the TGF-βsuperfamily,has multi-functional biological activities in regulating cell growth,differentiation,apoptosis and migration,and is an important mediator of renal fibrosis.However,the mechanism of renal fibrosis induced by TGF-β1 remains unclear.Our research group has screened out lnc RNA uc.412 in the previous experiment.This study aims to explore whether TGF-β1 can up-regulate the expression of lnc RNA uc.412 through the Smads pathway,changes the phenotype of glomerular mesangial cells and increases the synthesis of extracellular matrix,thereby inducing the renal fibrosis,and providing a new idea for the clinical prevention and treatment of chronic kidney disease(CKD).Methods:Smad3 phosphorylation specific inhibitors SIS3(1 umol/L)and TGF-β1(10ng/m L)were selected for experiments based on pre-experimental studies.The rat glomerular mesangial cells were divided into 4 groups and intervented for 24 h: the control group was treated with culture medium;the TGF-β1 group was treated with TGF-β1;the TGF-β1+SIS3 group was treated with TGF-β1 and SIS3;the SIS3 group was treated with SIS3.The expression levels of Collagen 1,α-SMA and p-Smad3 protein were detected by western blotting.Real-Time PCR was used to detect the expression level of lnc RNA uc.412.HBZY-1 cells were transfected by lentivirus lnc RNA uc.412.The Collagen 1 and α-SMA protein levels were detected by western blotting and the Collagen 1 and α-SMA m RNA levels were detected by Real-Time PCR.Results:The western blotting results showed that compared with the control group,p-Smad3,Collagen 1,α-SMA protein levels were increased in TGF-β1 group(P<0.05),compared with TGF-β1 group,the expression of p-Smad3,Collagen 1,and α-SMA protein in TGF-β1+ SIS3 group were decreased(P<0.05).Lnc RNA uc.412 overexpressed mesangial cell model was successfully constructed.Compared with the no-load control group,the overexpression of lnc RNA uc.412 upregulated the expression of α-SMA and Collagen 1 protein significantly(P<0.05).The real-time PCR results showed that compared with the control group,the expression level of lnc RNA uc.412 was significantly up-regulated(P<0.0001),compared with TGF-β1 group,the expression level of lnc RNA uc.412 in TGF-β1+ SIS3 group was significantly down-regulated(P<0.0001).Lnc RNA uc.412 overexpressed mesangial cell model was successfully constructed.Compared with the no-load control group,the overexpression of lnc RNA uc.412 upregulated the expression of α-SMA and Collagen 1 m RNA significantly(P<0.05).Conclusions:1.TGF-β1 can up-regulate the expression of lnc RNA uc.412 through the Smads signaling pathway,and inhibiting the Smads signaling pathway can down-regulate lnc RNA uc.412.2.The increased expression of lnc RNA uc.412 can change the phenotype of glomerular mesangial cells and increase the synthesis of extracellular matrix,thereby inducing renal fibrosis,which provides a new target for the effective treatment of chronic kidney disease.