Cadmium exposure on renal cell apoptosis,autophagy,oxidative damage and TGF-β1/Smad3/miRNAs signaling pathways
Objective To investigate the effects of Cd on apoptosis,autophagy,oxidative damage and TGF-β1/Smad3/miRNAs signaling pathway in HEK293 cells.Methods HEK293 cells were treated with CdCl2 for 6,12 and 24 h(model),and the cells without any treatment were set as the blank group(control).Cell proliferation was detected by MTT,apoptosis was detected by Annexin V-FITC/PI double staining,SOD activity was detected by WST-1 method,autophagy was detected by staining kit,the protein expression levels of TGF-β1 and Smad3 were detected by WB,and the expression of miR-21 was detected by qRT-PCR.On this basis,HEK293 cells with TGF-beta blockers SIS3 1/Smad3 pathway(CdCl2+SIS3 group),miR-NC group(CdCl2+miR-NC),and miR-21 inhibitor(CdCI2+miR-21 inhibi-tor,group)with 48 h after different methods for treatment of cadmium exposure HEK293 cells apoptosis,autophagy,oxidative damage,change,and TGF-beta 1/Smad3/miRNAs signal path expression level of relevant factors.Results At the same dose of CdCl2,the growth rate of HEK293 cells(model group)showed a time-dependent change,and the growth rate was significantly slower.Model group after cadmium exposure cell apoptosis rate(83.09+5.42)％,auto-phagy rate(77.16+8.91)％and TGF-beta 1,the amount of S mad3 protein expression were significantly higher than that of control group,model group cells SOD activity was significantly higher than that of control group(P＜0.05).Af-ter blocking the TGF-β1/Smad3 signaling pathway,the apoptosis rate of SIS3 group was(51.32±5.32)％,the autoph-agy rate was(43.52±2.46)％,the expression of TGF-β1 and Smad3 protein was lower than that of the model group,and the SOD activity was higher than that of the model group(P＜0.05).Cadmium exposure by miR-21 in HEK293 cells express,after compared with NC group,miR group-21 inhibitor apoptosis rate(46.18+4.95)％,autophagy rate(40.56+6.42)％,TGF-beta 1,the amount of Smad3 protein expression were significantly decline,SOD activity was significantly increased(P＜0.05).Conclusion Cadmium exposure induces oxidative damage,autophagy and apoptosis in renal cells in a time-dependent manner.Environmental cadmium exposure can increase the expression of TGF-β1/Smad3 related miRNA-21 and lead to renal function damage,and miRNA-21 may play a role in the process of autoph-agy,apoptosis and oxidative damage of renal cells induced by exposure through the TGF-β1/Smad3 pathway.
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