P2X4对帕金森病模型大鼠黑质中α突触核蛋白表达的影响  

EFFECT OF P2X4 ON EXPRESSION OFα-SYNUCLEIN IN THE SUBSTANTIA NIGRA OF A RAT MODEL OF PARKINSON'S DISEASE

作　　者：韩婧洋 张晓娜 房金妮 花菲菲 张晨[1] 谢安木[1] HAN Jingyang;ZHANG Xiaona;FANG Jinni;HUA Feifei;ZHANG Chen;XIE Anmu(Departmient of Neurology,The Affiliated Hospital of Qingdao Univcriy,Qingdao 266100,China)

机构地区：[1]青岛大学附属医院神经内科,山东青岛266100

出　　处：《精准医学杂志》2018年第1期62-66,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81571225)

摘　　要：目的探讨P2X4对6-羟基多巴胺(6-OHDA)诱导的帕金森病(PD)模型大鼠黑质中α突触核蛋白(α-synuclein)表达的影响。方法将120只雄性Wistar大鼠随机分为对照组、6-OHDA组、空载病毒阴性对照(P2X4-NC)组、目的基因阴性对照(siRNA-P2X4)组、P2X4-NC+6-OHDA组、siRNA-P2X4+6-OHDA组6组,每组20只大鼠。6-OHDA组、对照组只进行6-OHDA或等量生理盐水黑质定位注射,P2X4-NC组、siRNA-P2X4组分别用脑立体定位仪将空载阴性病毒或携带抑制P2X4表达的目的慢病毒定向注射于大鼠左侧黑质,P2X4-NC+6-OHDA组、siRNA-P2X4+6-OHDA组经黑质定位注射慢病毒2周后,于左侧黑质定位注射6-OHDA。4周后大鼠断头取脑,采用Western blot方法检测大鼠左侧黑质中P2X4、α-synuclein、半胱氨酸蛋白水解酶1(caspase-1)蛋白的表达,采用免疫组织荧光染色法检测各组大鼠左侧黑质中酪氨酸羟化酶(TH)阳性神经元数量的变化。结果与P2X4-NC+6-OHDA组比较,siRNA-P2X4+6-ODHA组P2X4、α-synuclein、caspase-1蛋白的表达量均明显减少(P<0.01),大鼠左侧黑质中TH阳性多巴胺能神经元数量明显增多(P<0.01)。结论抑制P2X4表达可下调黑质中α-synuclein的表达,对多巴胺能神经元有一定保护作用。Objective To investigate the effect of P2X4 on the expression ofα-synuclein in the substantia nigra of a rat model of Parkinson's disease(PD)induced by 6-hydroxydopamine(6-OHDA).Methods A total of 120 male Wiar ratswere equally and randomly divided into six groups:control group.6-OHDA group.P2X4-NC group,siRNA-P2X4 group.P2X4-NC+6-OHDA group,and siRNA-P2X4|6-OHDA group.The 6-OHDA group and the control group were given 6-OHDA and an equal volumie of normial saline,respectively,inthe left substantia nigra by stereotactic injection.The P2X4-NCgroup and the siRNA-P2X4 group were given negative control lentivirus and lentivirus of smiall interference RNA targeting P2X4,respectively,in the lett substantia nigra by stereotactic injection.The P2X4-NC|6-OHDA group and the siRNA-P2X4+6-OHDA group were given 6-OHDA in the lett substantia nigra by stereotactic injection at twoweeks after the treatments for the P2X4-NC group and the siR-NA-P2X4 group,respectively.Fourweeks later,the rats were sacrificed and the brain tissues were collected.The protein levels of P2X4,a-synuclein,and caspase-1 weremeasured by Western blot.The number of tyrosine hydroxylase(TH)-positive neurons in the left substantia nigra was measured by immunofluorescence staining.Compared with the P2X4-NC+6-OHDA group,the siRNA-P2X4+6-ODHA group had significantly lower protein levels of P2X4,α-synuclein,and caspase-1 and signifi-cantly more TH-positive neurons in the left substantia nigra(P<0.01).Conclusion The inhibition of P2X4 expression can down-regulate the expression ofα-synuclein in the substantia nigra and has a certain protective effect on dopaminergic neurons in PD rats.

关 键 词：受体 嘌呤能P2 帕金森病 突触核蛋白 半胱氨酸天冬氨酸蛋白酶1 大鼠 WISTAR 

分 类 号：R742.5[医药卫生—神经病学与精神病学]