Inhibitory effects of special siRNA targeting TLR4 gene on the apoptosis of renal proximal tubule epithelial cells induced by hypoxia-reoxygenation

Objective To study the inhibition of apoptosis of proximal tubule epithelial cells(PTECs) induced by hypoxia-reoxygenation injury by siRNA targeting TLR4 gene via the RNAi mechanisms.MethodsPTECs of Wistar rats were cultured in six-well plates and randomly divided into group A(normal group),group B(hypoxia-reoxygenation+TLR4-siRNA transfected group),group C(hypoxia-reoxygenation),group D(hypoxia-reoxygenation+con-siRNA transfected group).Group B,C,D were cultured in hypoxia condition for 3 h followed by reoxygenation for 24 h.RT-PCR and western blot were used to detect the expression of mRNA and protein of TLR4.Flow cytometry(FCM)was used to detect apoptosis of PTECs.ELISA was used to test the level of TNF-α in the supernatants.ResultsAfter hypoxia-reoxygenation,the expression of TLR4 mRNA and protein increased significantly in B,C and D group compared with A group(P0.01),the levels of TNF-α in the three groups increased significantly compared with A group(P0.01).The apoptosis cells in B,C and D increased significantly compared with A(P0.05).While the expression of TLR4 mRNA,TLR4 protein,the level of TNF-α in the supernatants and the apoptosis cells in B group decreased significantly compared with C and D group(P0.01);every measurement indicator was similar in D and C group(P0.05).ConclusionsSpecial siRNA targeting TLR4 gene could effectively prevent the apoptosis of PTECs and inflammatory reaction induced by hypoxia-reoxygenation.