沉默P2X4受体减少帕金森病细胞模型的凋亡  

Silencing P2X4 receptor reduces apoptosis in Parkinson’s disease cell model

作　　者：云强 董雪佳 刘雅虹 王智光[3] 张春雨[3] 江名芳[3] YUN Qiang;DONG Xue-jia;LIU Ya-hong;WANG Zhi-guang;ZHANG Chun-yu;JIANG Ming-fang(Department of Neurosurgery,Inner Mongolia Autonomous Region People’s Hospital,Hohhot 010020,Inner Mongolia Autonomous Region,China;Graduate School,Inner Mongolia Medical University,Hohhot 010058,Inner Mongolia Autonomous Region,China;Department of Neurology,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010059,Inner Mongolia Autonomous Region,China)

机构地区：[1]内蒙古自治区人民医院神经外科,呼和浩特010020 [2]内蒙古医科大学研究生院,呼和浩特010058 [3]内蒙古医科大学附属医院神经内科,呼和浩特010059

出　　处：《海军军医大学学报》2022年第12期1423-1428,共6页Academic Journal of Naval Medical University

基　　金：内蒙古医科大学附属医院博士启动金资助项目(NYFY BS 2018);内蒙古自治区科技计划项目(2021GG0233)。

摘　　要：目的 研究P2X4受体(P2X4R)在帕金森病(PD)细胞模型中的作用。方法 用6-羟基多巴胺(6-OHDA)处理人神经母细胞瘤细胞SH-SY5Y,建立PD细胞模型并检测P2X4R的表达。在SH-SY5Y细胞中沉默P2X4R,采用CCK-8法检测细胞活性,流式细胞术检测细胞凋亡情况,蛋白质印迹法检测凋亡因子和泛连接蛋白1(PANX1)的表达。在PD模型中沉默P2X4R、抑制P2X4R、过表达PANX1,用蛋白质印迹法检测细胞中Toll样受体2(TLR2)蛋白的表达。在PD模型细胞中过表达TLR2,检测细胞活性、细胞凋亡情况和凋亡因子表达。结果 100、150μmol/L 6-OHDA处理后PD模型细胞中P2X4R蛋白和mRNA表达水平均高于对照组(P均<0.05)。P2X4R沉默后PD模型细胞凋亡减少、细胞存活率提高,caspase 3、cleaved caspase 3及PANX1蛋白表达降低(P均<0.05)。沉默P2X4R和抑制P2X4R后PD模型细胞中TLR2蛋白表达均降低(P均<0.05),过表达PANX1后TLR2蛋白表达降低(P<0.05),过表达TLR2后caspase 3和cleaved caspase 3蛋白表达降低(P均<0.05)、细胞存活率提高。结论 P2X4R可能通过调控PANX1及TLR2表达减少PD模型细胞凋亡。Objective To study the role of P2X4 receptor(P2X4R) in a cell model of Parkinson’s disease(PD).Methods Treat human neuroblastoma cells SH-SY5Y with 6-hydroxydopamine(6-OHDA) to establish a Parkinson’s disease cell model and detect the expression of P2X4R. P2X4R was silenced in SH-SY5Y cells, cell viability was detected by cell counting kit 8(CCK-8), cell apoptosis was detected by flow cytometry, and the expression of apoptosis factors and pannexin 1(PANX1) was detected by Western blotting. Western blotting was used to detect the expression of Toll-like receptor 2(TLR2)protein when P2X4R was silenced, P2X4R was inhibited and PANX1 was overexpressed. TLR2 was overexpressed in PD model cells, and cell viability, apoptosis, and apoptotic factor expression were examined. Results P2X4R protein and mRNA expression levels in PD model cells treated with 100 and 150 μmol/L 6-OHDA were higher than those in control group(all P<0.05). After P2X4R silence, the apoptosis of PD model cells was significantly decreased and cell survival rate was significantly increased;and the protein expression of cysteine aspartic acid specific protease(caspase) 3, cleaved caspase 3 and PANX1was significantly decreased(all P<0.05). The expression of TLR2 protein in PD model cells was decreased after silencing P2X4R and inhibiting P2X4R(both P<0.05). The expression of TLR2 protein was significantly decreased after PANX1overexpression(P<0.05). After TLR2 overexpression, the protein expression of caspase 3 and cleaved caspase 3 was significantly decreased(both P<0.05), and the cell survival rate was increased. Conclusion P2X4R may reduce apoptosis of PD model cells by regulating PANX1 and TLR2 expression.

关 键 词：帕金森病 P2X4受体 泛连接蛋白1 TOLL样受体2 细胞凋亡 

分 类 号：R742.5[医药卫生—神经病学与精神病学]