Hyperosmotic Stress-Induced TRPM2 Channel Activation Stimulates NLRP3 Inflammasome Activity in Primary Human Corneal Epithelial Cells
Hyperosmotic Stress-Induced TRPM2 Channel Activation Stimulates NLRP3 Inflammasome Activity in Primary Human Corneal Epithelial Cells
复制标题
高渗应激诱导的 TRPM2 通道激活刺激原代人角膜上皮细胞中的 NLRP3 炎症小体活性
DOI：
10.1167/iovs.18-23965
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发表时间：
2018-07-01
影响因子：
4.4
通讯作者：
Chen, Wei
中科院分区：
文献类型：
--
作者：
Zheng, Qinxiang;Tan, Qiufan;Chen, Wei
目的.本研究的目的是确定高渗或氧化应激是否诱导NLRP 3炎性小体激活并通过瞬时受体电位melastatin 2（TRPM 2）激活原代人角膜上皮细胞（PHECs）增加生物活性IL-1 β分泌。实时PCR、Western印迹和免疫荧光染色分别用于评估TRPM 2和NLRP 3、ASC、caspase-1和IL-1 β的mRNA和蛋白表达水平。CCK-8测定评价细胞活力。施加高渗500 mOsm和氧化0.5 mM H2 O2应激。TRPM 2表达用TRPM 2抑制剂、20 μ M N-（对戊基肉桂酰基）邻氨基苯甲酸（ACA）或TRPM 2 siRNA敲低来抑制。在高渗培养基中，TRPM 2、NLRP 3、ASC、caspase-1和IL-1 β基因和蛋白表达水平在4小时后升高（P
PURPOSE. The purpose of this study was to determine whether either a hyperosmotic or oxidative stress induces NLRP3 inflammasome activation and increases in bioactive IL-1 beta secretion through transient receptor potential melastatin 2 (TRPM2) activation in primary human corneal epithelial cells (PHCECs).METHODS. Real-time PCR, Western blots, and immunofluorescent staining were used to evaluate TRPM2 and NLRP3, ASC, caspase-1, and IL-1 beta mRNA and protein expression levels, respectively. A CCK-8 assay evaluated cell viability. Hyperosmotic 500 mOsm and oxidative 0.5 mM H2O2 stresses were imposed. TRPM2 expression was inhibited with a TRPM2 inhibitor, 20 lM N-(p-amylcinnamoyl) anthranilic acid (ACA), or TRPM2 siRNA knockdown.RESULTS. In the hypertonic medium, TRPM2, NLRP3, ASC, caspase-1, and IL-1 beta gene and protein expression levels rose after 4 hours (P