P2X4受体对帕金森病大鼠脑黑质中脑源性神经营养因子表达的影响  

Effect of P2X4 receptor on the expression of brain-derived neurotrophic factor in substantia nigra of rats with Parkinson′s disease

作　　者：魏丽萍 赵俊杰 谷晓林 WEI Liping;ZHAO Junjie;GU Xiaolin(Department of Neurology,Jinan City People′s Hospital,Jinan 271199,Shandong Province,China)

机构地区：[1]济南市人民医院神经内科,山东济南271199

出　　处：《新乡医学院学报》2023年第3期207-212,共6页Journal of Xinxiang Medical University

摘　　要：目的探讨P2X4受体(P2X4R)对帕金森病(PD)大鼠脑黑质中脑源性神经营养因子(BDNF)表达的影响。方法采用随机数字表法将120只Wistar雄性大鼠随机分为对照组、6-羟基多巴胺(6-OHDA)组、P2X4组、P2X4-阴性对照(NC)组、P2X4+6-OHDA组、P2X4-NC+6-OHDA组,每组20只。对照组大鼠于左侧黑质注射2μL含体积分数0.02%抗坏血酸的生理盐水;6-OHDA组大鼠于左侧黑质注射2μL 6-OHDA构建PD模型;P2X4组大鼠于左侧黑质注射2μL携带过表达P2X4基因的慢病毒;P2X4-NC组大鼠于左侧黑质注射2μL空载阴性病毒;P2X4+6-OHDA组大鼠于左侧黑质注射2μL携带过表达P2X4基因的慢病毒,1周后注射2μL 6-OHDA;P2X4-NC+6-OHDA组大鼠于左侧黑质注射2μL空载阴性病毒,1周后注射2μL 6-OHDA。采用免疫荧光染色法检测各组大鼠左侧黑质中酪氨酸羟化酶(TH)阳性多巴胺能神经元数量,Western blot法检测各组大鼠左侧黑质中P2X4R、BDNF蛋白表达。结果与对照组比较,6-OHDA组大鼠左侧黑质中TH阳性多巴胺能神经元数量和BDNF蛋白相对表达量显著降低,P2X4R蛋白相对表达量显著升高(P<0.05)。与P2X4-NC组比较,P2X4-NC+6-OHDA组大鼠左侧黑质中TH阳性多巴胺能神经元数量及BDNF蛋白相对表达量显著降低,P2X4R蛋白相对表达量显著升高(P<0.05)。P2X4组大鼠左侧黑质中P2X4R蛋白相对表达量显著高于P2X4-NC组(P<0.05)。与P2X4组比较,P2X4+6-OHDA组大鼠左侧黑质中TH阳性多巴胺能神经元数量和BDNF蛋白相对表达量显著降低,P2X4R蛋白相对表达量显著升高(P<0.05)。与6-OHDA组比较,P2X4+6-OHDA组大鼠左侧黑质中TH阳性多巴胺能神经元数量和BDNF蛋白相对表达量显著降低,P2X4R蛋白相对表达量显著升高(P<0.05)。与P2X4-NC+6-OHDA组比较,P2X4+6-OHDA组大鼠左侧黑质中TH阳性多巴胺能神经元数量和BDNF蛋白相对表达量显著降低,P2X4R蛋白相对表达量显著升高(P<0.05)。结论PD大鼠过表达P2X4R可降低BDNF的表达水平,减�Objective To investigate the effect of P2X4 receptor(P2X4R)on the expression of brain-derived neurotrophic factor(BDNF)in substantia nigra of Parkinson′s disease(PD)rats.Methods A total of 120 male Wistar rats were randomly divided i nto the control group,6-hydroxydopamine(6-OHDA)group,P2X4 group,P2X4-negative control(NC)group,P2X4+6-OHDA group and P2X4-NC+6-OHDA group,with 20 rats in each group.The rats in the control group were injected with 2μL saline containing 0.02%ascorbic acid by volume in the left substantia nigra;the rats in the 6-OHDA group were injected with 2μL 6-OHDA to establish PD model in the left substantia nigra;the rats in the P2X4 group were injected with 2μL lentivirus carrying overexpression of P2X4 gene in the left substantia nigra;the rats in the P2X4-NC group were injected with 2μL negative virus in the left substantia nigra;the rats in the P2X4+6-OHDA group were injected with 2μL lentivirus carrying overexpression of P2X4 gene in the left substantia nigra,and injected with 2μL 6-OHDA after one week;the rats in the P2X4-NC+6-OHDA group were injected with 2μL empty negative virus in the left substantia nigra,and injected with 2μL 6-OHDA after one week.The number of tyrosine hydroxylase(TH)positive dopaminergic neurons in left substantia nigra of rats in each group was detected by immunofluorescence staining,and the expressions of P2X4R and BDNF were detected by Western blot method.Results Compared with the control group,the number of TH positive dopaminergic neurons and the relative expression of BDNF protein of rats in the 6-OHDA group were significantly decreased,while the relative expression of P2X4R protein was significantly increased(P<0.05).Compared with the P2X4-NC group,the number of TH positive dopaminergic neurons and the relative expression of BDNF protein of rats in the P2X4-NC+6-OHDA group were significantly decreased,while the relative expression of P2X4R protein was significantly increased(P<0.05).Compared with the 6-OHDA group,the relative expression of P2X4R prot

关 键 词：帕金森病 P2X4受体 脑源性神经营养因子 多巴胺能神经元 

分 类 号：R742.5[医药卫生—神经病学与精神病学]