Atilano-Roque Amandla, Aleksunes Lauren M, Joy Melanie S
Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado, Aurora, CO, United States.
Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, NJ, United States.
Toxicol Lett. 2016 Sep 30;259:52-59. doi: 10.1016/j.toxlet.2016.07.021. Epub 2016 Jul 29.
Cisplatin is prescribed for the treatment of solid tumors and elicits toxicity to kidney tubules, which limits its clinical use. Nuclear factor erythroid 2-related factor 2 (Nrf2, NFE2L2) is a critical transcription factor that has been shown to protect against kidney injury through activation of antioxidant mechanisms. We aimed to evaluate the ability of short-term treatment with the Nrf2 activator bardoxolone methyl (CDDO-Me) to protect against cisplatin-induced kidney cell toxicity. Cell viability was assessed in human kidney proximal tubule epithelial cells (hPTCs) exposed to low, intermediate, and high cisplatin concentrations in the presence and absence of CDDO-Me, administered either prior to or after cisplatin. Treatment with cisplatin alone resulted in reductions in hPTC viability, while CDDO-Me administered prior to or after cisplatin exposure yielded significantly higher cell viability (17%-71%). Gene regulation (mRNA expression) studies revealed the ability of CDDO-Me to modify protective pathways including Nrf2 induced detoxifying genes [GCLC (increased 1.9-fold), NQO1 (increased 9.3-fold)], and an efflux transporter [SLC47A1 (increased 4.5-fold)] at 12h. Protein assessments were in agreement with gene expression. Immunofluorescence revealed localization of GCLC and NQO1 to the nucleus and cytosol, respectively, with CDDO-Me administered prior to or after cisplatin exposure. The findings of enhanced cell viability and increased expression of detoxifying enzymes (GCLC and NQO1) and the multidrug and toxin extrusion protein 1 (MATE1) efflux transporter (SLC47A1) in hPTCs exposed to CDDO-Me, suggest that intermittent treatment with CDDO-Me prior to or after cisplatin exposure may be a promising approach to mitigate acute kidney injury.
顺铂被用于治疗实体瘤，但会引发肾小管毒性，这限制了其临床应用。核因子红细胞2相关因子2（Nrf2，NFE2L2）是一种关键的转录因子，已被证明可通过激活抗氧化机制来预防肾损伤。我们旨在评估短期使用Nrf2激活剂巴多昔芬甲酯（CDDO-Me）预防顺铂诱导的肾细胞毒性的能力。在存在和不存在CDDO-Me的情况下，对暴露于低、中、高顺铂浓度的人肾近端小管上皮细胞（hPTCs）的细胞活力进行评估，CDDO-Me在顺铂之前或之后给药。单独使用顺铂治疗会导致hPTC活力降低，而在顺铂暴露之前或之后给予CDDO-Me可使细胞活力显著提高（17%-71%）。基因调控（mRNA表达）研究表明，CDDO-Me能够在12小时时改变包括Nrf2诱导的解毒基因[GCLC（增加1.9倍）、NQO1（增加9.3倍）]和一种外排转运蛋白[SLC47A1（增加4.5倍）]在内的保护途径。蛋白质评估与基因表达一致。免疫荧光显示，在顺铂暴露之前或之后给予CDDO-Me时，GCLC和NQO1分别定位于细胞核和细胞质。在暴露于CDDO-Me的hPTCs中，细胞活力增强、解毒酶（GCLC和NQO1）和多药及毒素外排蛋白1（MATE1）外排转运蛋白（SLC47A1）表达增加的结果表明，在顺铂暴露之前或之后间歇性使用CDDO-Me可能是减轻急性肾损伤的一种有前景的方法。
