Abstract: Necrostatin-1 (Nec-1) inhibits necroptosis and is usually regarded as having no effect on other cell deaths. Here, this study explored whether the addition of Nec-1 has an effect on cell death induced by simulated ischemia injury in rat tubular cell line NRK-52E. In addition, we also investigated the mechanism of Nec-1 attenuates cell death in this renal ischemia model. The NRK-52E cells were incubated with TNF-α + antimycinA (TA) for 24 h with or without Nec-1. Cell death was observed under fluorescent microscope and quantified by flow cytometry. Cell viabilities were detected by MTT assay. The protein expression of dynamin-related protein 1 (Drp1) was detected by Western blotting and immunofluorescence assay. Increased cell death in simulated ischemia injury of NRK-52E cells were markedly attenuated in the Nec-1 pretreated ischemia injury group. Meanwhile, cell viability was significantly improved after using Nec-1. In addition, we also observed that the protein expression of Drp1, a mediator of mitochondrial fission, was significantly increased in simulated ischemia injury group. Increased Drp1 expression in the ischemia injury group can be abolished by Nec-1 or Drp1-knock down, accompanied with decreased cell death and improved cell viabilities. These results suggest that Nec-1 may inhibit cell death induced by simulated ischemia injury in the rat tubular cell line NRK-52E through decreased Drp1 expression.
Keywords: Necrostatin-1; cell death; renal ischemia injury; dynamin-related protein 1
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Zhang, L.; Jiang, F.; Chen, Y.; Luo, J.; Liu, S.; Zhang, B.; Ye, Z.; Wang, W.; Liang, X.; Shi, W. Necrostatin-1 Attenuates Ischemia Injury Induced Cell Death in Rat Tubular Cell Line NRK-52E through Decreased Drp1 Expression. Int. J. Mol. Sci. 2013, 14, 24742-24754.
Zhang L, Jiang F, Chen Y, Luo J, Liu S, Zhang B, Ye Z, Wang W, Liang X, Shi W. Necrostatin-1 Attenuates Ischemia Injury Induced Cell Death in Rat Tubular Cell Line NRK-52E through Decreased Drp1 Expression. International Journal of Molecular Sciences. 2013; 14(12):24742-24754.
Zhang, Li; Jiang, Fen; Chen, Yuanhan; Luo, Jialun; Liu, Shuangxin; Zhang, Bin; Ye, Zhiming; Wang, Wenjian; Liang, Xinling; Shi, Wei. 2013. "Necrostatin-1 Attenuates Ischemia Injury Induced Cell Death in Rat Tubular Cell Line NRK-52E through Decreased Drp1 Expression." Int. J. Mol. Sci. 14, no. 12: 24742-24754.