P2X4过表达对帕金森病模型大鼠脑内细胞凋亡的作用  

The effect of overxpression of P2X4 on cell apoptosis in a rat model of Parkinson Disease

作　　者：刘佳琳 花菲菲 张英英[1] 侯炳辉[1] 谢安木[1] LIU Jialin;HUA Feifei;ZHANG Yingying;HOU Binghui;XIE Anmu(Department of Neurology, The Affiliated Hospital of Qingdao University, No 16, Jiangsu Road, Qingdao 266003, China.)

机构地区：[1]青岛大学附属医院神经内科

出　　处：《中国神经精神疾病杂志》2018年第3期134-138,共5页Chinese Journal of Nervous and Mental Diseases

基　　金：国家自然基金资助项目(编号:81571225)

摘　　要：目的探讨过表达P2X4基因对6-羟基多巴(6-hydroxydopamine,6-OHDA)诱导的帕金森病(Parkinson disease,PD)大鼠模型中脑内细胞凋亡的影响。方法 120只Wistar雄性大鼠随机分为6组(n=20):(1)对照组,(2)6-OHDA组,(3)单纯目的基因(RNA-P2X4)组,(4)目的基因(RNA-P2X4)+6-OHDA组,(5)单纯阴性病毒(P2X4-NC)组,(6)阴性病毒(P2X4-NC)+6-OHDA组。根据分组情况分别于大鼠左侧黑质定向注射携带目的基因的慢病毒或阴性病毒和(或)6-OHDA或等量生理盐水。免疫荧光法检测各组黑质中多巴胺能神经元细胞数;Western blot法分别检测各组黑质中P2X4R、caspase-1及NLRP3蛋白表达水平。结果与P2X4-NC+6-OHDA组比较,RNA-P2X4+6-OHDA组P2X4R(1.099±0.05569 vs.0.7821±0.02008,P=0.0003)、NLRP3(0.9875±0.01932 vs.0.6645±0.01747,P<0.0001)、caspase-1(0.9948±0.01788 vs.0.8276±0.04543,P<0.0001)蛋白表达量均显著升高,差异有统计学意义。结论 P2X4在PD的发病中发挥重要作用,P2X4激活后可增加NLRP3炎性体和caspase-1的表达,促进炎症及凋亡反应,有潜在的神经损害作用。Objective To investigate the effect of overexpression of P2X4 gene on cell apoptosis in 6-OHDA（6-hydroxydopamine, 6-OHDA） induced PD rat model. Methods One hundred twenty male Wistar rats were randomly divided into 6 groups： 6-OHDA group, control group, objective RNA-P2X4 ＋ 6-OHDA group, target gene RNA-P2X4 group, P2X4-NC ＋ 6-OHDA group and negative virus P2X4-NC only group. Lentivirus or negative virus carrying the target gene and or 6-OHDA or the same amount of physiological saline were injected into the left substantia nigra of rats according to the groups. The number of dopaminergic neurons in substantia nigra was measured by immunofluorescence method. The expression levels of P2X4R, caspase-1 and NLRP3 in substantia nigra were detected by Western blot assay.Results Compared with P2X4-NC ＋ 6-OHDA group, the expression of protein P2X4R（1.099 ± 0.05569 vs. 0.7821 ±0.02008, P=0.0003）, NLRP3（0.9875 ± 0.01932 vs. 0.6645 ± 0.01747. P〈0.0001）, caspase-1（0.9948 ± 0.01788 vs.0.8276 ± 0.04543, P 〈0.0001） increased significantly in RNA-P2X4 ＋6-OHDA group. Conclusion overexpression of P2X4 gene can increase the expression of NLPR3 and caspase-1 protein and promote apoptosis in PD rats.

关 键 词：过表达P2X4 帕金森病 CASPASE-1 NLRP3 

分 类 号：R742.5[医药卫生—神经病学与精神病学]