Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation.
Knocking down TRPM2 expression reduces cell injury and NLRP3 inflammasome activation in PC12 cells subjected to oxygen-glucose deprivation.
复制标题
敲低 TRPM2 表达可减少氧糖剥夺的 PC12 细胞中的细胞损伤和 NLRP3 炎性体激活。
DOI：
10.4103/1673-5374.282271
复制
发表时间：
2020-11
影响因子：
6.1
通讯作者：
Feng X
中科院分区：
文献类型：
--
作者：
Pan T;Zhu QJ;Xu LX;Ding X;Li JQ;Sun B;Hua J;Feng X
瞬时受体电位Melastatin 2(TRPM2)是一种重要的离子通道，是治疗脑缺血损伤的潜在靶点。然而，目前尚不清楚减少TRPM2的表达是否有助于修复脑损伤，如果是的话，这一过程背后的机制是什么。本研究探讨了降低TRPM2表达对缺氧缺糖致嗜铬细胞瘤(PC12)细胞损伤的保护作用。将编码TRPM2 shRNAs的质粒导入PC12细胞，在无糖的低氧条件下孵育8h，复氧24 h。用流式细胞仪检测细胞凋亡、线粒体膜电位、活性氧水平和细胞内钙离子水平。用荧光定量逆转录聚合酶链式反应和Western blotting检测C-X-C基序趋化因子配体2(CXCL2)、NACHT、LRR、PYD结构域蛋白3(NALP3)和caspase-1的相对表达。TRPM2-shRNA+OGD组细胞凋亡率、线粒体膜电位、活性氧水平和细胞内钙水平均低于OGD组。综上所述，这些结果表明，TRPM2基因敲除可能通过抑制细胞凋亡和降低氧化应激水平、线粒体膜电位、细胞内钙浓度和NLRP3炎性小体激活来减轻OGD诱导的神经元损伤。
Transient receptor potential melastatin 2 (TRPM2) is an important ion channel that represents a potential target for treating injury caused by cerebral ischemia. However, it is unclear whether reducing TRPM2 expression can help repair cerebral injury, and if so what the mechanism underlying this process involves. This study investigated the protective effect of reducing TRPM2 expression on pheochromocytoma (PC12) cells injured by oxygen-glucose deprivation (OGD). PC12 cells were transfected with plasmid encoding TRPM2 shRNAS, then subjected to OGD by incubation in glucose-free medium under hypoxic conditions for 8 hours, after which the cells were allowed to reoxygenate for 24 hours. Apoptotic cells, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels were detected using flow cytometry. The relative expression of C-X-C motif chemokine ligand 2 (CXCL2), NACHT, LRR, and PYD domain–containing protein 3 (NALP3), and caspase-1 were detected using fluorescence-based quantitative reverse transcription-polymerase chain reaction and western blotting. The rates of apoptosis, mitochondrial membrane potentials, reactive oxygen species levels, and cellular calcium levels in the TRPM2-shRNA + OGD group were lower than those observed in the OGD group. Taken together, these results suggest that TRPM2 knockdown reduces OGD-induced neuronal injury, potentially by inhibiting apoptosis and reducing oxidative stress levels, mitochondrial membrane potentials, intracellular calcium concentrations, and NLRP3 inflammasome activation.