Key Genes Of Renal Interstitial Fibrosis And Tubular Atrophy

Objective By establishing aristolochic acid A-induced epithelialmesen-chymal transformation model of human proximal tubular epithelial cells,i-t was verified that the molecular RAC2 screened by bioinformatics is rel-ated to the epithelial-mesenchymal transformation of renal tubular epitheli-al cells.Method(1)Download the microarray data set from the GEO database.Then,select and identify differentially expressed genes(DEGs),accomplish functional beneficiation analysis,construct a PPI network,and carry out modular analysis of DEGs.Comprehensively considering the current literature research related to candidate genes,public database data related to kidney disease and KEGG analysis,we selected the gene RAC2 as the molecule for further verification research.(2)Human proximal tubular epithelial cell lines were cultured in vitro.According to the relevant literature,the experimental groups were set up:(0,5,10,20,40 ug/ml)with different concentrations of AA,and the human proximal tubular epithelial cell model was established with different concentrations of aristolochic acid A.CCK-8 method was used to detect the damage of human proximal tubular epithelial cells treated with different concentrations of AA(0,5,10,20,40 ug/ ml)for 48 hours.It was Real time PCR which was used to detect the mRNA comparative expression of E-cadherin,?-smooth muscle actin,vimentin and Rac2.(3)Add RAC2 inhibitor EHT 1864 to the human proximal tubular epithelial cell model induced by aristolochic acid A,and then detect Ecadherin and ? by real-time quantitative PCR method(Real-time PCR method)-Relative expression of smooth muscle actin and RAC2 mRNA,CCK8 method was used to detect cell proliferation activity.Results(1)After 48 hours of intervention with different concentrations of aristolochic acid A,the activity of human proximal renal tubular epithelial cells was inhibited in varying degrees,displaying a dose-dependent survival inhibition phenomenon,the difference was statistically significant.(2)The results of realtime fluorescent quantitative PCR displayed that contrast to the normal control group,the expression of E-cadherin mRNA was significantly reduced,?-smooth muscle actin,vimentin and Rac2 mRNA were significantly increased,and the difference was statistically significant.(3)After adding EHT 1864,the expression of E-cadherin mRNA was higher than that of the experimental group,the expression of ?-SMA mRNA was lower than that of the experimental group,there was no significant change in the expression of RAC2 mRNA,and the cell survival rate was higher than that of the experimental group.Conclusion Aristolochic acid A can cause proliferation inhibition of human proximal tubular epithelial cells and induce cell damage;human proximal tubular epithelial cells suffer from aristolochic acid A damage,forming an epithelial mesenchymal transformation effect;After adding RAC2 inhibitor EHT1864 to inhibit RAC2,the effect of aristolochic acid A-induced epithelial mesenchymal transformation was improved,and RAC2 may be related to epithelial mesenchymal transformation.