Effect Of Bailing Extract On Cyclosporine-induced Epithelial-mesenchymal Transition And TGF-?1/Smads Signaling Pathway In Renal Tubular Epithelial Cells

Objective: To investigate the effect of Bailing extract on CsA-induced renal tubular epithelial cell transdifferentiation(EMT)and the effect of Bailing extract on renal tubular EMT-related TGF-?1/Smads signaling protein.Methods: 1.Normal rat renal tubular epithelial cell line(NRK-52E)was divided into normal control group and different drugs(cyclosporine serum TGF-?1 inhibitor group,Bailing extract original solution),CCK8 detects cell viability.2.NRK-52 E cell cultures were divided into 6 groups: group A: blank control group;group B: Cyclosporine group;group C: Bailing extract group;group D:Cyclosporine +TGF-?1 inhibitor group;Group E: Cyclosporine + Bailing extract group;Group F:Cyclosporine + Bailing extract + TGF-?1 inhibitor group,The expression of ?-smooth muscle actin(?-SMA),TGF-?1,Smad2,and Smad7 mRNA and protein were detected by RT-PCR,western-blot and immunohistochemistry.Results: 1.The survival rate of cyclosporine A-induced renal tubular epithelial cells was significantly decreased.After 24 hours of treatment with TGF-?1 receptor inhibitor or Bailing extract liquid,the cell viability was higher in cyclosporine A group than in cyclosporine A group(P<0.05),the survival rate of the Bailing extract intervention group was higher than that of the TGF-?1 receptor inhibitor group(P<0.05),and the optimal concentrations of the TGF-?1 receptor inhibitor and the Bailing extract solution were 10?mol/L and 0.5 mg/L,respectively.2.?-SMA,Smad2 mRNA and protein:Compared with A and C groups,the mRNA and protein expressions of B,D,E and F groups were significantly up-regulated,and the difference was statistically significant(P<0.05),The upregulation of B group wassignificantly higher than that of D,E,F groups,but there was no significant difference between C and A groups(P>0.05).Comparison between D,E,and F groups,there was no significant difference in protein expression between D and E groups(P>0.05).The expression of protein in E group was up-regulated compared with F group(P<0.05).3.TGF-?1 mRNA and protein:Compared with A and C groups,the expression of mRNA and protein in B,D,E and F groups increased significantly(P<0.05),and the expression of mRNA and protein in B group was significantly higher than that in D,E and F groups.There was no significant difference between C and A group(P>0.05).There was no significant difference between D,E and F groups(P>0.05),while the difference between E and F groups was statistically significant(P<0.05).4.Smad7 mRNA and protein:The mRNA and protein expressions of B,D,E and F groups were lower than those of A and C groups(P<0.05),especially B group.The down-regulation of B group was especially significant,but there was no statistical difference between A and C groups(P>0.05).There was no significant difference between D and E groups(P>0.05),but there was significant difference between D and F,E and F groups(P<0.05).Conclusions: Bailing extract can improve cyclosporine induced renal tubular epithelial mesenchymal transition,and some me chanisms can be achieved by regulating the expression of related molecular proteins in the TGF-?1/Smads signaling pathway.