Colistin-Induced Nephrotoxicity in Mice Involves the Mitochondrial, Death Receptor, and Endoplasmic Reticulum Pathways
Colistin-Induced Nephrotoxicity in Mice Involves the Mitochondrial, Death Receptor, and Endoplasmic Reticulum Pathways
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粘菌素诱导的小鼠肾毒性涉及线粒体、死亡受体和内质网途径
DOI：
10.1128/aac.00070-14
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发表时间：
2014-07-01
影响因子：
4.9
通讯作者：
Xiao, Xilong
中科院分区：
文献类型：
--
作者：
Dai, Chongshan;Li, Jichang;Xiao, Xilong
肾毒性是粘菌素的剂量限制因素，但确切机制尚不清楚。本研究旨在探讨线粒体、死亡受体和内质网途径在粘菌素诱导的肾毒性中的作用。小鼠静脉内给予7.5或15 mg粘菌素/kg体重/天（通过3分钟输注，分为两次给药），持续7天。测定肾功能、氧化应激和细胞凋亡。研究了涉及线粒体、死亡受体和内质网途径的代表性生物标志物，并检查了涉及凋亡和自噬的关键标志物。治疗7 d后，血尿素氮、血肌酐和丙二醛显著升高，肾脏超氧化物歧化酶（SOD）和过氧化氢酶活性降低。检测到急性肾小管坏死和线粒体功能障碍，并通过DNA片段化、聚（ADP-核糖）聚合酶（PARP-1）裂解、8-羟基脱氧鸟苷（8-OHdG）增加和半胱天冬酶（caspase-8、-9和-3）激活来表征粘菌素诱导的细胞凋亡。很明显，粘菌素诱导的细胞凋亡涉及线粒体途径（Bcl-2下调和细胞色素C [cytC]和Bax上调）、死亡受体途径（Fas、FasL和Fas相关死亡结构域[FADD]上调）和内质网途径（Grp 78/Bip、ATF 6、GADD 153/CHOP和caspase-12上调）。在15-mg/kg/d粘菌素组中，细胞周期蛋白依赖性激酶2（CDK 2）和磷酸化JNK（p-JNK）的表达显著增加（P < 0.05），而在7.5-mg/kg/d粘菌素组中，观察到大量的自噬溶酶体和经典的自噬。Beclin-1和LC 3B蛋白的Western blot结果表明，自噬可能在粘菌素诱导的肾毒性中起保护作用。总之，这是第一个研究表明，所有三个主要的细胞凋亡途径和自噬参与粘菌素诱导的肾毒性。
Nephrotoxicity is the dose-limiting factor for colistin, but the exact mechanism is unknown. This study aimed to investigate the roles of the mitochondrial, death receptor, and endoplasmic reticulum pathways in colistin-induced nephrotoxicity. Mice were intravenously administered 7.5 or 15 mg of colistin/kg of body weight/day (via a 3-min infusion and divided into two doses) for 7 days. Renal function, oxidative stress, and apoptosis were measured. Representative biomarkers involved in the mitochondrial, death receptor, and endoplasmic reticulum pathways were investigated, and the key markers involved in apoptosis and autophagy were examined. After 7-day colistin treatment, significant increase was observed with blood urea nitrogen, serum creatinine, and malondialdehyde, while activities of superoxide dismutase (SOD) and catalase decreased in the kidneys. Acute tubular necrosis and mitochondrial dysfunction were detected, and colistin-induced apoptosis was characterized by DNA fragmentation, cleavage of poly(ADP-ribose) polymerase (PARP-1), increase of 8-hydroxydeoxyguanosine (8-OHdG), and activation of caspases (caspase-8, -9, and -3). It was evident that colistin-induced apoptosis involved the mitochondrial pathway (downregulation of Bcl-2 and upregulation of cytochrome C [cytC] and Bax), death receptor pathway (upregulation of Fas, FasL, and Fas-associated death domain [FADD]), and endoplasmic reticulum pathway (upregulation of Grp78/Bip, ATF6, GADD153/CHOP, and caspase-12). In the 15-mg/kg/day colistin group, expression of the cyclin-dependent kinase 2 (CDK2) and phosphorylated JNK (p-JNK) significantly increased (P < 0.05), while in the 7.5-mg/kg/day colistin group, a large number of autophagolysosomes and classic autophagy were observed. Western blot results of Beclin-1 and LC3B indicated that autophagy may play a protective role in colistin-induced nephrotoxicity. In conclusion, this is the first study to demonstrate that all three major apoptosis pathways and autophagy are involved in colistin-induced nephrotoxicity.