Experimental Study Of Centella Asiatica Particles On TGF-β1-induced In Vitro Cultured Tubular Epithelial Cells Of E-cadherin、FSP-1、FN Expression

Background: The incidence of chronic kidney disease is about 10.8%[1].The patients who eventually suffer from the end-stage renal failure,will have to rely on renal replacement therapy(including renal transplantation,hemodialysis and peritoneal dialysis)to live on.Renal tubule interstitial fibrosis is a common pathway and the main pathological basis of all kinds of kidney disease which would be progressed to end-stage renal failure.Characteristics of tubule interstitial fibrosis are the deposition of extracellular matrix(ECM),infiltration of inflammatory cells and fibroblast accumulation,but its mechanism is not clear[2].In recent years,studies have shown that,the epithelial mesenchymal transdifferentiation(EMT)is the key pathogenesis of tubule interstitial fibrosis.The study have shown that renal tubular epithelial cells transform into myofibroblasts after EMT and then led to excessive deposition of ECM,resulting in the occurrence of tubular interstitial fibrosis[3].Objective: In this study,we investigated the effect of Centella asiatica particles on TGF-β1-induced in vitro cultured tubular pithelial cells of the E cadherin、FN、FSP-1m RNA and protein of FN、FSP-1 expression,and explore the possible mechanism of Centella asiatica in the prevention and treatment of tubule interstitial fibrosis,in order to provide a theoretical basis for the clinical application of Centella asiatica.Methods: We chose Fosinopril(Monopril)group as the control group,rat proximal tubular epithelial cells were randomly divided into 6 groups: 1.Normal control group;2.TGF-β1-induced group;3.Centella asiatica low-dose group;4.Centella asiatica middle-dose group;5.Centella asiatica high-dose group;6.Monopril group.The application of realtime fluorescence quantitative PCR(RT-PCR)to detect the mRNA of E-cadherin、FN、FSP-1,application of western-blot detect FN、FSP-1 would be taken after these cells have grown for 48 hours.The experiment was repeated three times in each group.Results:1.Compared with the control group,TGF-β1-induced group arranged disorderly,which transfrom cobblestone epithelial cells into the spindle myofibroblast.The Centella asiatica high-dose group cell morphology did not change significantly and the change of monopril group in morphology cell was similar with the high-dose group.2.Compared with the control group,TGF-β1-induced group down-regulate the expression of E-cadherin m RNA,and increase FN,FSP-1m RNA expression(P<0.05);Compared with the TGF-β1-induced group,the Centella asiatica group and the monopril group was reduced in the expression of FN 、 FSP-1m RNA and increased in the expression of E-cadherin m RNA(P<0.05);Compared with monopril group,the expression of E-cadherin、FN、FSP-1m RNA in Centella asiatica high-dose group was similar(P>0.05);3.Compared with the control group,the protein expression of FN.FSP-1 significantly increased in TGF-β1-induced group(P<0.05);Compared with the TGF-β1-induced group,the protein expression of FN 、 FSP-1 in centella asiatica high-dose group and monopril group significantly decreased(P<0.05).The change of monopril group is similar with centella asiatica high-dose group(P>0.05).Conclusion: 1.Centella asiatica suppresse the expression of FN、FSP-1 and promote the expression of E-cadherin in TGF-β1-induced in vitro cultured tubular epithelial cells;2.There was no difference between high dose centella asiatica group and fosinopril groupin in the effect of suppressing the expression of FN、FSP-1 and promoting the expression of E-cadherin;3.The anti-TIF effect of the centella asiatica was correlated with the dosage;4.Centella asiatica may has the effect of anti-TIF by suppressing the epithelial mesenchymal transdifferentiation.