The protective effect of EGF-activated ROS in human corneal epithelial cells by inducing mitochondrial autophagy via activation TRPM2
The protective effect of EGF-activated ROS in human corneal epithelial cells by inducing mitochondrial autophagy via activation TRPM2
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egf激活的ROS通过激活TRPM2诱导线粒体自噬对人角膜上皮细胞的保护作用
DOI：
10.1002/jcp.29597
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发表时间：
2020-02-21
影响因子：
5.6
通讯作者：
Jin, Xiuming
中科院分区：
文献类型：
--
作者：
Huo, Yanan;Chen, Wei;Jin, Xiuming
氧化应激是某些眼表疾病的主要发病机制。我们的前期研究表明表皮生长因子（EGF）激活的活性氧（ROS）可以保护人角膜上皮细胞（HCE）免受损伤。本研究旨在探讨氧化应激和线粒体自噬在划痕损伤HCE细胞中的作用及其机制。分别进行CCK-8测定、EdU测定、蛋白质印迹分析、伤口愈合测定和流式细胞术以确定细胞活力、增殖、蛋白质表达、细胞凋亡和细胞内ROS水平。结果表明，EGF可通过上调ROS促进划痕损伤的HCE细胞的损伤修复并抑制细胞凋亡（**p < .01，*p < .001）。EGF还诱导线粒体自噬，减轻线粒体损伤。有趣的是，线粒体自噬抑制剂和线粒体分裂抑制剂1（MDIVI-1）与EGF的组合可以降低细胞增殖、活力和ROS水平（*p < .05，**p < .01，*p < .001）。使用ROS抑制剂N-乙酰基-L-半胱氨酸的治疗废除了EGF治疗后线粒体膜电位的增加。（*p < .05）。总之，这些发现表明EGF在HCE损伤修复中起重要作用，并且可以通过激活TRPM 2诱导线粒体自噬来激活ROS以保护免受HCE损伤。
Oxidative stress is a major pathogenesis of some ocular surface diseases. Our previous study demonstrated that epidermal growth factor (EGF)-activated reactive oxygen species (ROS) could protect against human corneal epithelial cell (HCE) injury. In the present study, we aimed to explore the role and mechanisms of oxidative stress and mitochondrial autophagy in HCE cells subjected to scratch injury. CCK-8 assays, EdU assays, Western blot analysis, wound-healing assays, and flow cytometry were conducted to determine cell viability, proliferation, protein expression, cell apoptosis, and intracellular ROS levels, respectively. The results showed that EGF could promote damage repair and inhibit cell apoptosis in scratch injured HCE cells by upregulating ROS (**p < .01, ***p < .001). EGF also induced mitochondrial autophagy and alleviated mitochondrial damage. Interestingly, the combination of the mitochondrial autophagy inhibitor and mitochondrial division inhibitor 1 (MDIVI-1) with EGF could reduce cell proliferation, viability, and the ROS level (*p < .05, **p < .01, ***p < .001). Treatment using the ROS inhibitor N-acetyl- l-cysteine abrogated the increase in mitochondrial membrane potential after EGF treatment. (*p < .05). Taken together, these findings indicated that EGF plays an important role in HCE damage repair and could activate ROS to protect against HCE injury by inducing mitochondrial autophagy via activation of TRPM2.