Wu Yu, Zhang Yang, Wang Ling, Diao Zongli, Liu Wenhu
1. Department of Nephrology, Beijing Friendship Hospital, Capital Medical University, No. 95 Yong An Road, Xi Cheng District, Beijing 100050, China ; 2. Department of Nephrology, The First People's Hospital of Xuzhou, No. 19 Zhongshan North Road, Xuzhou 221002, Jiangsu, China.
3. Department of Anesthesiology, Xuzhou Medical College, Xuzhou 221004, Jiangsu, China.
Int J Med Sci. 2015 Aug 1;12(8):655-67. doi: 10.7150/ijms.12460. eCollection 2015.
Acute kidney injury (AKI) is a systemic inflammatory response syndrome associated with poor clinical outcomes. No treatments effective for AKI are currently available. Thus, there is an urgent need of development of treatments effective for AKI. Autophagy, an intracellular proteolytic system, is induced in renal cells during AKI. However, whether autophagy is protective or injurious for AKI needs to be clearly clarified. We addressed this question by pharmacological inhibition of autophagy using a mouse model of lipopolysaccharide (LPS) induced-AKI. We found that autophagy was induced in renal cortex of mice during LPS-induced AKI as reflected by a dose-and time-dependent increased accumulation of light chain 3-II (LC3-II), the common marker of autophagy, compared to that of control group; 2) the occurrence of intensive, punctate and increased immunohistochemical staining image of LC3-II in renal cortex; 3) the significant increase in the expression levels of Beclin-1, another key marker of autophagy; 4) the significantly increased levels of plasma urea and serum creatinine and 5) the significant increase in autophagagosome area ratio. We observed that 3-methyladenine (3-MA), a pharmacological inhibitor of autophagy, blocked autophagy flux, alleviated AKI and protected against LPS-induced AKI. LPS triggered kidney inflammation by activation of the canonical NF-κB pathway. This route can be modulated by autophagy. Activation of the canonical NF-κB pathway was reduced in 3-MA+LPS as compared to that in LPS-treated group of mice. Mice pretreated with 3-MA before exposure to LPS showed a reduction in p65 phosphorylation, resulting in the accumulation of ubiquitinated IκB. In conclusion, impairment of autophagy ameliorates LPS-induced inflammation and decreases kidney injury. The accumulation of ubiquitinated IκB may be responsible for this effect.
急性肾损伤(AKI)是一种与不良临床结局相关的全身炎症反应综合征。目前尚无对AKI有效的治疗方法。因此,迫切需要开发对AKI有效的治疗方法。自噬是一种细胞内蛋白水解系统,在AKI期间在肾细胞中被诱导。然而,自噬对AKI是保护性的还是有害的,需要明确阐明。我们通过使用脂多糖(LPS)诱导的AKI小鼠模型对自噬进行药理学抑制来解决这个问题。我们发现,与对照组相比,在LPS诱导的AKI期间,小鼠肾皮质中自噬被诱导,这表现为自噬的常见标志物轻链3-II(LC3-II)的积累呈剂量和时间依赖性增加;2)肾皮质中LC3-II出现密集、点状且增加的免疫组化染色图像;3)自噬的另一个关键标志物Beclin-1的表达水平显著增加;4)血浆尿素和血清肌酐水平显著升高;5)自噬体面积比显著增加。我们观察到,自噬的药理学抑制剂3-甲基腺嘌呤(3-MA)阻断了自噬通量,减轻了AKI,并对LPS诱导的AKI起到保护作用。LPS通过激活经典的NF-κB途径引发肾脏炎症。这条途径可被自噬调节。与LPS处理的小鼠组相比,3-MA+LPS组中经典NF-κB途径的激活减少。在暴露于LPS之前用3-MA预处理的小鼠显示p65磷酸化减少,导致泛素化的IκB积累。总之,自噬受损可改善LPS诱导的炎症并减轻肾损伤。泛素化的IκB的积累可能是这种作用的原因。