Study Of Emodin On TGF-?1-induced Epithelial-mesenchymal Transition And Mitofusin2 In Human Renal Tubular Epithelial Cells

Background:Emodin is one of the effective active ingredients in Chinese medicine rhubarb.It has antibacterial,anti-inflammatory,immune regulation,renal function improvement,and anti-fibrotic effects.TGF-?1 is a renal fibrotic factor,which can induce renal tubular epithelial cells EMT and promote the development of renal fibrosis.Recent studies have pointed out that Mitofusin2(Mfn2)and mitochondrial function are closely related to the occurrence of chronic kidney disease.So,the effect of emodin on EMT and mitochondrial function of human renal tubular epithelial cells induced by TGF-?1 is worthy of further study.Objectives:To investigate the effects of emodin on TGF-?1-induced human renal tubular epithelial cells(HK-2)epithelial-mesenchymal transition and Mfn2.Methods:1.Using the CCK8 method to detect the proliferation-toxic effects of different concentrations of EM on HK-2 cells;2.The experiment cells were divided into blank control group,TGF-?1 group and TGF-?1+EM group.The emodin intervention group was treated with different concentrations of emodin(10,20 and 40?mol/L)for 30 min.After that,the TGF-?1 group and the emodin intervention group were simultaneously stimulated with TGF-?1 at a concentration of 10 ng/mL for 48 h.The cells were then harvested and the expressions of E-cadherin,?-smooth muscle actin(a-SMA),connective tissue growth factor(CTGF),Mfn2 mRNA and protein were detected by RT-qPCR and western blot,respectively.Results:1.Effects of emodin at different concentrations on the proliferation-toxic effects of HK-2 cells10,20,40,80,and 160?mol/L concentrations of EM all had inhibitory effects on the proliferation of HK-2 cells,and the inhibitory effects were more pronounced at 80 and 160?mol/L concentrations.2.Effect of TGF-?1 on E-cadherin,a-SMA,CTGF and Mfn2 mRNA and proteins in HK-2 cellsCompared with the blank control group,the expression of a-SMA,CTGF,Mfn2 mRNA and protein increased and the expression of E-cadherin mRNA and protein decreased in HK-2 cells after TGF-?1 stimulation.3.Effect of emodin on E-cadherin,a-SMA,CTGF and Mfn2 mRNA and proteins in HK-2 cells after TGF-?1 stimulationCompared with the TGF-?1 group,the expression of a-SMA,CTGF,and Mfn2 mRNA and protein decreased,and the expression of E-cadherin mRNA and protein increased in dose-dependent manner.Conclusions:Emodin attenuates TGF-?1-induced epithelial-mesenchymal transition in human renal tubular epithelial cells,which may ameliorate renal fibrosis by reducing the expression of TGF-?1-induced Mfn2.