Effects Of Angiotensinii On Necroptosis In Rat Renal Tubular Epithelial Cells

Objective:To study the effects of Angiotension II(Ang II) on necroptosis in rat renal tubular epithelial cells (NRK-52E).Methods:The small interfering RNA (siRNA)targeted to the RIP3mRNA was synthesized in vitro and was transfected by the siRNA-mate transfection reagents into NRK-52E. Knockdown of the target molecule, RIP3, was monitored by Western Blot analysis. The cell viability was evaluated by MTT assay. The cell apoptosis rate was determined by flow cytometer(FCM). The intracellular ROS levels was assessed by a fluorescent microplate reader. The mRNA and protein expression of RIP1 RIP3 was detected by qPCR and Western Blotting.Results:Sequence specific siRNAs of RIP3 effectively down regulated the protein levels of RIP3 compared with the control group(P<0.01). Compared with the control group, Ang? group significantly decreased the cell viability and increased the cell apoptotic rate, the ROS levels, the expression of RIP1 and RIP3 (P< 0.01).Compared with Ang? group, Z-VAD increased the cell viability and decreased the number of apoptotic cells, the ROS levels(P<0.01). Z-VAD could up-regulate the expression of RIP1 and RIP3 of Ang? group(P<0.01). Nec-1 or RIP3siRNA significantly increased the cell viability and suppressed the number of necrotic cells, the ROS levels, the expression of RIP 1 and RIP3 (P< 0.01). No significant difference on the apoptosis(P>0.05).Conclusion:AngII can induce the necroptosis of NRK-52E through the production of RIP 1, RIP3 and ROS.