Protective effects of isorhamnetin on N2a cell against endoplasmic reticulum stress-induced injury is mediated by PKC epsilon
Protective effects of isorhamnetin on N2a cell against endoplasmic reticulum stress-induced injury is mediated by PKC epsilon
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PKC epsilon 介导异鼠李素对 N2a 细胞内质网应激损伤的保护作用
DOI：
10.1016/j.biopha.2017.06.062
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发表时间：
2017
影响因子：
7.5
通讯作者：
Lu Huiqiang
中科院分区：
文献类型：
--
作者：
Qiu Lingyu;Ma Yurong;Luo Yougen;Cao Zigang;Lu Huiqiang
内质网应激(ERS)诱导的细胞内钙超载和ROS爆发在细胞凋亡中起关键作用。蛋白激酶C epsilon(PKCε)参与调节钙离子和ROS产生的动态平衡。异鼠李素(Iso)作为一种ROS清除剂，能有效地抑制细胞凋亡，但其作用机制尚不清楚。本研究旨在探讨Iso是否能抑制内质网诱导的N2a细胞的凋亡，其保护作用与PKCε介导的钙稳态和抑制ROS有关。通过细胞存活率、细胞内钙离子水平、细胞凋亡率和活性氧(ROS)水平检测Iso对N2细胞ERS损伤的保护作用。免疫印迹法检测GRP78蛋白表达水平。结果表明，Iso通过抑制钙超载，减少ROS的产生，减少细胞凋亡，从而减轻ERS所致的损伤。此外，Iso还能促进PKCε的磷酸化，εV1-2(一种PKCε抑制剂)可显著减弱Iso对N2acellERS损伤的保护作用。综上所述，我们首次证明了Iso对N2a细胞的内质网损伤具有保护作用，这种保护作用至少部分是通过PKCε途径实现的。
Endoplasmic reticulum stress (ERS)-induced intracellular calcium (Ca2+) overload and ROS burst plays a critical role in apoptosis. Protein kinase C epsilon (PKCε) is involved in regulating the homeostasis of Ca2+and ROS production. isorhamnetin (Iso), as an ROS scavenger, effectively inhibit apoptosis, but the mechanism is still unclear. This study was to investigate whether Iso can inhibit ERS-induced apoptosis in N2a cells, and the protective effects are involved in PKCε-mediated Ca2+homeostasis and inhibition of ROS. The effects of Iso against ERS injury inN2acells were detected by cell viability, the levels of Ca2+, apoptosis and reactive oxygen species (ROS). The protein GRP78 expression levels were measured by western blot assay. The results showed that Iso can reduce ERS-induced injury by inhibiting Ca2+overload, reducing the generation of ROS and decreasing apoptosis. In addition, Iso can promote PKCε phosphorylation, and εV1-2 (a PKCε inhibitor) drastically attenuated the protective effects of Iso against ERS injury inN2acells. In conclusion, we firstly demonstrated that Iso can elicit protective effects against ERS injury in N2a cells and these effects are mediated at least in part via PKCε pathway.