侧脑室注射腺病毒介导的GDNF基因对帕金森病的保护作用

Neuroprotective effect of adenovirus-mediated GDNF gene in mice models of Parkinsons disease

目的探讨腺病毒介导的神经胶质细胞源性神经营养因子(GDNF)基因脑内转移对帕金森病(PD)的保护作用。方法采用C57Bl/6小鼠1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-henyl-l,2,3,6-tetrahydropyridine,M PTP)法建立PD模型,随机分为重组GDNF腺病毒(Ad-GDNF)实验组和对照组。Ad-GDNF实验组将Ad-GDNF定向注射至侧脑室,对照组将无GDNF基因的腺病毒定向注射至侧脑室,并测量体质量,进行行为学实验评分,行中脑黑质酪氨酸羟化酶(TH)免疫组化染色,高压液相色谱-电化学仪(HPLC-ECD)检测纹状体多巴胺(DA)、5-羟色胺(5-HT)及5-羟吲哚乙酸(5-HIAA)含量,采用ELISA、RT-PCR法检测Ad-GDNF在中脑的表达。结果侧脑室注射病毒后2周,Ad-GDNF实验组体质量、行为学实验得分、黑质酪氨酸羟化酶(TH)阳性细胞、纹状体DA含量均显著高于对照组(P<0.05);Ad-GDNF实验组在中脑内有过表达,中脑GDNF含量约是对照组2倍。结论侧脑室注射腺病毒介导的GDNF基因可减轻1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-henyl-l,2,3,6-tetrahydropyridine,M PTP)诱发的小鼠DA能神经元进行性变性,保护DA能神经元,提示这一手段在PD保护性治疗方面具有一定的应用价值。

Objective To investigate the neuroprotective effect of adenovirus-mediated glial cell line-derived neurotrophic factor（ GDNF） gene intracerebrally transferred on the treatment of Parkinsons disease（ PD）. Methods PD models w ere established by intraperitoneal injection of 1-methyl-4-henyl-l,2,3,6-tetrahydropyridine（ M PTP） [30 mg /（ kg · d） in C57 Bl /6 mice. The models w ere then divided into Ad-GDNF experimental group and control group. Ad-GDNF w as injected into the lateral ventricle of mice in the experimental group. Negative virus w as injected into the lateral ventricle of mice in the control group. The body w eight of all mice w as measured. The neuroprotective effect of Ad-GDNF w as evaluated w ith behavioral tests,immune histochemical assay of the tyrosine hydroxylase（ TH）-positive neurons in the substance nigra and measurement of monoamine level in the striatum. The expression of the exogenous GDNF gene in the brain w as detected w ith ELISA and RT-PCR. Results Compared w ith control group,the experimental group show ed improved motor functions（ P〈0. 01） and less loss of body w eight（ P〈0. 05）,better survival of TH-positive cells（ P〈0. 05）,higher DA levels（ P〈0. 05） after tw o w eeks of intracerebroventricular injection. The exogenousGDNF gene w as efficiently over expressed in the experimental group. GDNF protein level in midbrain of the experimental group w as tw ice of the control group. Conclusion Intracerebroventricular injection of adenovirus mediated GDNF gene significantly can protect the dopaminergic neurons of nigrostriatal system from M PTP-induced injury and is conducive in the treatment of Parkinsons disease.