P2X4R过表达对帕金森病模型鼠脑黑质中白介素-1β、α突触核蛋白及多巴胺能神经元的影响  

The Effect of P2X4R Overexpression on the Number of IL-1β、α-synuclein and Dopaminergic Neurons in the Substantia Nigra of PD Rat Model

作　　者：魏丽萍[1,2] 薛莉 许文帅[1] 谢安木[1] WEI Li-ping;XUE Li;XU Wen-shuai;XIE Anmu(Department of Neurology,the Affiliated Hospital of Qingdao University,Qingdao 266003,China;Department of Neurology,Jinan People5 s Hospital,Jinan 271199,China;Department of Medicine,the Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院神经科,青岛266003 [2]济南市人民医院神经内科,济南271199 [3]青岛大学附属医院病案室,青岛266003

出　　处：《中国临床神经科学》2019年第6期617-623,共7页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金资助项目(编号:81571225)

摘　　要：目的探讨使用慢病毒过表达P2X4R嘌呤受体(P2X4R)对帕金森病(PD)模型大鼠脑黑质中白介素-1β(IL-1β)和α突触核蛋白的表达及多巴胺能神经元数量的影响。方法 Wistar雄性大鼠,体质量200~250 g,随机分为6组(均n=20)。经立体定向定位左侧黑质,对照组:注入含0. 02%维生素C的生理盐水;PD组:注入6-羟基多巴胺(6-OHDA);目的基因阴性对照组:注入携带过表达P2X4R的慢病毒;空载病毒阴性对照组:注入空载病毒;目的基因阴性对照+PD组:注入携带过表达P2X4R的慢病毒1周后再注射6-OHDA;空载病毒阴性对照+PD组:注入空载病毒1周后再注射6-OHDA。模型判定成功后处死大鼠取左侧黑质,Western blot法检测各组黑质P2X4R、IL-1β、α突触核蛋白表达水平;免疫荧光染色法检测各组黑质酪氨酸羟化酶(TH)阳性多巴胺能神经细胞数的变化。结果①与PD组和空载病毒阴性对照+PD组比较,目的基因阴性对照+PD组P2X4R(P <0. 001)、IL-1β(P <0. 001)和α突触核蛋白(P <0. 01)相对表达量增加;②与PD组和空载病毒阴性对照+PD组比较,目的基因阴性对照+PD组TH阳性多巴胺能神经元数量明显减少(P <0. 001)。结论慢病毒介导P2X4R过表达可上调PD模型大鼠黑质中IL-1β和α-突触核蛋白表达水平、减少TH阳性多巴胺能神经细胞数;小胶质细胞的炎症反应增强,α-突触核蛋白沉积增加,对多巴胺能神经元有一定的损伤作用。Aim To investigate the effect of lentivirus(LV-P2X4R)overexpressing P2X4R receptor(P2X4R)on the release of IL-1β and a-synuclein and the number of dopaminergic neurons in Parkinson's disease(PD)rat model.Methods The male Wistar rats weighing 200 to 250 grams were randomly divided into 6 groups,with 20 rats in each group.Stereotactic left midbrain substantia nigra.The control group was injected with saline containing 0.02%vitamin C.The 6-OHDA group was injected with 6-OHDA・The RNA-P2X4R group was injected with lentivirus carrying overexpressed P2X4R.The P2X4RNC group was injected with airborne virus.The RNA-P2X4R+6-OHDA group was injected with lentivirus carrying overexpressed P2X4R first,and then 6-OHDA a week later.The P2X4R-NC+6-OHDA group was injected with airborne virus first,and then 6-OHDA a week later.After the model was successfully determined,the rats were decapitated to collect the left substantia nigra tissues.The protein expression levels of P2X4R,IL-lβ and a-synuclein in each group were detected using Western blotting.Meanwhile,changes in the TH positive neuron number were detected by immunofluorescence staining.Results(J)Compared with the 6-OHDA group and P2X4R-NC+6-OHDA group,the expression of P2X4R、IL・1β and a-synuclein increased in RNA-P2X4R+6-OHDA group(PP2X4R<0.001,P1L_lβ<0.001,Pa_syn<0.01).②Compared with the 6-OHDA group and P2X4R-NC+6-OHDA group,the numbers of TH positive dopaminergic neurons in RNA-P2X4R+6-OHDA group were notably reduced(P<0.001).Conclusion Lentivirus-mediated P2X4R overexpression can upregulate the expression of IL-1β and a-synuclein and decrease the numbers TH positive dopaminergic neurons,which means enhancing the inflammatory response of microglia can increase the deposition of a・synuclein and damage dopaminergic neurons to a certain extent.

关 键 词：帕金森病 P2X4R瞟吟受体 过表达 白介素-IB a突触核蛋白 

分 类 号：R742.5[医药卫生—神经病学与精神病学]