[1]魏丽萍,赵俊杰,谷晓林.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[J].新乡医学院学报,2023,40(3):207-212.[doi:10.7683/xxyxyxb.2023.03.002]
　WEI Liping,ZHAO Junjie,GU Xiaolin.Effect of P2X4 receptor on the expression of brain-derived neurotrophic factor in substantia nigra of rats with Parkinson′s disease[J].Journal of Xinxiang Medical University,2023,40(3):207-212.[doi:10.7683/xxyxyxb.2023.03.002]
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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
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
40卷
期数:
2023年3期
页码:
207-212
栏目:
基础研究
出版日期:
2023-03-05

文章信息/Info

Title:
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
（济南市人民医院神经内科，山东　济南　271199）
Author(s):
WEI Liping; ZHAO Junjie; GU Xiaolin
(Department of Neurology,Jinan City People′s Hospital,Jinan 271199,Shandong Province,China )
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
Keywords:
Parkinson′s disease; P2X4 receptor; brain-derived neurotrophic factor; dopaminergic neuron
分类号:
R742.5
DOI:
10.7683/xxyxyxb.2023.03.002
文献标志码:
A
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2 μL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2 μL 6-OHDA构建PD模型；P2X4组大鼠于左侧黑质注射2 μL携带过表达P2X4基因的慢病毒；P2X4-NC组大鼠于左侧黑质注射2 μL空载阴性病毒；P2X4+6-OHDA 组大鼠于左侧黑质注射2 μL携带过表达P2X4基因的慢病毒，1周后注射2 μL 6-OHDA；P2X4-NC+6-OHDA组大鼠于左侧黑质注射2 μL空载阴性病毒，1周后注射2 μL 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，减弱BDNF的神经保护作用，加剧多巴胺能神经元的丢失，进而导致PD的进展。
Abstract:
ObjectiveTo 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.MethodsA total of 120 male Wistar rats were randomly divided into 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 2 μL saline containing 0.02% ascorbic acid by volume in the left substantia nigra;the rats in the 6-OHDA group were injected with 2 μL 6-OHDA to establish PD model in the left substantia nigra;the rats in the P2X4 group were injected with 2 μL lentivirus carrying overexpression of P2X4 gene in the left substantia nigra;the rats in the P2X4-NC group were injected with 2 μL negative virus in the left substantia nigra;the rats in the P2X4+6-OHDA group were injected with 2 μL lentivirus carrying overexpression of P2X4 genein the left substantia nigra,and injected with 2 μL 6-OHDA after one week;the rats in the P2X4-NC+6-OHDA group were injected with 2 μL empty negative virus in the left substantia nigra,and injected with 2 μL 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 BDNFprotein 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 protein of rats in the P2X4 group was significantly higher than that in the P2X4-NC group（P<0.05）.Compared with the P2X4 group,the number of TH positive dopaminergic neurons and the relative expression of BDNF protein of rats in the P2X4+6-OHDA group were significantly decreased,while the relative expression of P2X4R protein was significantly increased（P<0.05）.The number of TH positive dopaminergic neurons and the relative expression of BDNF protein of rats in the P2X4+6-OHDA group were significantly decreased,while the relative expression of P2X4R protein was significantly increased（P<0.05）.Compared with the P2X4-NC+6-OHDA group,the number of TH positive dopaminergic neurons and the relative expression of BDNF protein of rats in the P2X4+6-OHDA group were significantly decreased,while the relative expressionof P2X4R protein was significantly increased（P<0.05）.ConclusionOverexpression of P2X4R in PD rats can reduce the expression level of BDNF,weaken the neuroprotective effect of BDNF,and aggravate the loss of dopaminergic neurons,thus leading to the progress of PD.

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更新日期/Last Update: 2023-03-05