Wang Xinhui, Parrish Alan R
Medical Pharmacology and Physiology, School of Medicine, University of Missouri, MA 415 Medical Sciences Building, One Hospital Drive, Columbia, MO, 65212, USA.
Apoptosis. 2015 Jul;20(7):921-9. doi: 10.1007/s10495-015-1129-x.
The aging kidney undergoes structural and functional alterations which make it more susceptible to drug-induced acute kidney injury (AKI). Previous studies in our lab have shown that the expression of α(E)-catenin is decreased in aged kidney and loss of α(E)-catenin potentiates AKI-induced apoptosis, but not necrosis, in renal tubular epithelial cells (NRK-52E cells). However, the specific apoptotic pathway underlying the increased AKI-induced cell death is not yet understood. In this study, cells were challenged with nephrotoxicant cisplatin to induce AKI. A ~5.5-fold increase in Fas expression in C2 (stable α(E)-catenin knockdown) relative to NT3 (non-targeted control) cells was seen. Increased caspase-8 and -9 activation was induced by cisplatin in C2 as compared to NT3 cells. In addition, decreased Bcl-2 expression and increased BID cleavage and cytochrome C release were detected in C2 cells after cisplatin challenge. Treating the cells with cisplatin, in combination with a Bcl-2 inhibitor, decreased the viability of NT3 cells to the same level as C2 cells after cisplatin. Furthermore, caspase-3/-7 activation is blocked by Fas, caspase-8, caspase-9 and pan-caspase inhibitors. These inhibitors also completely abolished the difference in viability between NT3 and C2 cells in response to cisplatin. These results demonstrate a Fas-mediated apoptotic signaling pathway that is enhanced by the age-dependent loss of α(E)-catenin in renal tubule epithelial cells.
衰老的肾脏会发生结构和功能改变,使其更容易受到药物诱导的急性肾损伤(AKI)。我们实验室之前的研究表明,老年肾脏中α(E)-连环蛋白的表达降低,α(E)-连环蛋白的缺失会增强肾小管上皮细胞(NRK-52E细胞)中AKI诱导的凋亡,但不会增强坏死。然而,AKI诱导的细胞死亡增加背后的具体凋亡途径尚不清楚。在本研究中,用肾毒性顺铂刺激细胞以诱导AKI。相对于NT3(非靶向对照)细胞,C2(稳定的α(E)-连环蛋白敲低)细胞中Fas表达增加了约5.5倍。与NT3细胞相比,顺铂在C2细胞中诱导了caspase-8和-9的激活增加。此外,在顺铂刺激后,C2细胞中检测到Bcl-2表达降低、BID切割增加和细胞色素C释放增加。用顺铂联合Bcl-2抑制剂处理细胞后,NT3细胞的活力降低至与顺铂处理后的C2细胞相同水平。此外,Fas、caspase-8、caspase-9和泛caspase抑制剂可阻断caspase-3/-7的激活。这些抑制剂还完全消除了NT3和C2细胞在对顺铂反应中活力的差异。这些结果表明,在肾小管上皮细胞中,Fas介导的凋亡信号通路因α(E)-连环蛋白的年龄依赖性缺失而增强。