Abstract:

Objective To explore the effects of angiotension Ⅱ (Ang Ⅱ) on the necroptosis of rat renal tubular epithelial cells (NRK-52E). Methods The small interfering RNA (siRNA) targeted at the RIP3 mRNA was synthesized in vitro and was transfected by the transfection reagent siRNA-mate to NRK-52E. The protein expression of RIP3 was detected by the Western blotting. The cell viability was evaluated by the MTT assay. The cell apoptosis rate was determined by the flow cytometer. The intracellular reactive oxygen species (ROS) level was detected by the fluorescent microplate reader. The mRNA and protein expressions of RIP1 and RIP3 were detected by the qPCR and Western blotting. Results Compared with the control group, the protein expression of RIP3 significantly decreased after RIP3 siRNA was transfected by NRK-52E (P<0.01). Compared with the control group, the cell viability of the AngⅡ group decreased, while the apoptotic rate, ROS level, and expressions of RIP1 and RIP3 significantly increased (P<0.01). Compared with the AngⅡ group, Z-VAD increased the cell viability, decreased the apoptotic rate and ROS level (P<0.01), and up-regulated the expressions of RIP1 and RIP3 (P<0.01). Nec-1 and RIP3 siRNA decreased the necrosis rate (P<0.01), had no significant effect on the apoptotic rate (P>0.05), and decreased the ROS level and expressions of expressions of RIP1 and RIP3 (P<0.01). Conclusion AngⅡ can induce the necroptosis of rat renal tubular epithelial cells, which is relevant to the generation of RIP1, RIP3 and ROS.

Key words: necroptosis, angiotensionⅡ, renal tubular epithelial cell, RIP1, RIP3, reactive oxygen species