Reactive oxygen species-initiated autophagy opposes aldosterone-induced podocyte injury
Reactive oxygen species-initiated autophagy opposes aldosterone-induced podocyte injury
复制标题
活性氧引发的自噬对抗醛固酮诱导的足细胞损伤
DOI：
10.1152/ajprenal.00409.2015
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发表时间：
2016-04-01
影响因子：
4.2
通讯作者：
Zhang, Aihua
中科院分区：
文献类型：
--
作者：
Bai, Mi;Che, Ruochen;Zhang, Aihua
有证据表明，醛固酮（Aldo）参与慢性肾脏疾病的发生和发展。本研究的目的是探讨自噬在醛类药物诱导足细胞损伤中的作用及其机制。在存在或不存在3-甲基腺嘌呤和N-乙酰半胱氨酸的情况下，用Aldo处理小鼠足细胞。通过检测膜联蛋白V结合物、凋亡小体、caspase-3活性和足细胞蛋白nephrin的改变来研究细胞凋亡。通过测量轻链3、p62、beclin-1和自噬相关基因5的表达来评估自噬。Aldo（10（-7）mol/l）诱导足细胞凋亡、自噬和nephrin蛋白表达下调，呈时间依赖性。3-甲基腺嘌呤和自噬相关基因5小干扰RNA预处理可抑制自噬，进一步促进乙醛诱导的细胞凋亡。此外，Aldo时间依赖性地增加了活性氧的产生，并且H2 O2（10（-4）mol/l）的应用显着增加了足细胞自噬。用N-乙酰半胱氨酸处理后，由Aldo或H2 O2诱导的自噬明显减弱，表明ROS在介导足细胞自噬形成中起关键作用。抑制ROS也可减轻Aldo诱导的足细胞损伤。总之，我们的研究结果表明，ROS触发的自噬对Aldo诱导的足细胞损伤起保护作用，靶向足细胞自噬可能代表治疗足细胞病的新治疗策略。
Evidence has demonstrated that aldosterone (Aldo) is involved in the development and progression of chronic kidney diseases. The purpose of the present study was to investigate the role of autophagy in Aldo-induced podocyte damage and the underlying mechanism. Mouse podocytes were treated with Aldo in the presence or absence of 3-methyladenine and N-acetylcysteine. Cell apoptosis was investigated by detecting annexin V conjugates, apoptotic bodies, caspase-3 activity, and alterations of the podocyte protein nephrin. Autophagy was evaluated by measuring the expressions of light chain 3, p62, beclin-1, and autophagy-related gene 5. Aldo (10(-7) mol/l) induced podocyte apoptosis, autophagy, and downregulation of nephrin protein in a time-dependent manner. Aldo-induced apoptosis was further promoted by the inhibition of autophagy via 3-methyladenine and autophagy-related gene 5 small interfering RNA pretreatment. Moreover, Aldo time dependently increased ROS generation, and H2O2 (10(-4) mol/l) application remarkably elevated podocyte autophagy. After treatment with N-acetylcysteine, the autophagy induced by Aldo or H2O2 was markedly attenuated, suggesting a key role of ROS in mediating autophagy formation in podocytes. Inhibition of ROS could also lessen Aldo-induced podocyte injury. Taken together, our findings suggest that ROS-triggered autophagy played a protective role against Aldo-induced podocyte injury, and targeting autophagy in podocytes may represent a new therapeutic strategy for the treatment of podocytopathy.