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目的探讨重组慢病毒介导的Persephin基因治疗对6-OHDA制备的帕金森病(PD)模型大鼠多巴胺能神经元的保护作用。方法将64只雄性Wistar大鼠随机分为正常组、6-OHDA组、空病毒+6-OHDA组、Persephin慢病毒预处理+6-OHDA组,并给予相应处理。6-OHDA注射21d后检测大鼠的旋转行为,免疫荧光检测黑质多巴胺能神经元数目,Western blot检测Persephin及酪氨酸羟化酶(TH)在脑内黑质区的表达。结果Persephin慢病毒黑质注射4周后,多巴胺能神经元成功表达Persephin蛋白(F=5.451,q=3.228~4.736,P<0.05)。6-OHDA注射21d后,Persephin慢病毒预处理+6-OHDA组旋转次数明显低于6-OHDA组和空病毒+6-OHDA组(F=7.551,q=4.228、5.336,P<0.05),多巴胺能神经元数目和TH表达水平显著高于6-OHDA组和空病毒+6-OHDA组(F=6.251、5.251,q=3.228~5.736,P<0.05)。结论 Persephin对6-OHDA制备的PD模型大鼠多巴胺能神经元具有保护作用。
Objective To investigate the protective effect of recombinant lentivirus-mediated Persephin gene therapy on dopaminergic neurons in Parkinson’s disease (PD) model rats induced by 6-OHDA. Methods Sixty-four male Wistar rats were randomly divided into 6 groups: normal group, 6-OHDA group, empty virus + 6-OHDA group and Persephin lentivirus + 6-OHDA group. The rotational behavior of rat was detected after 6-OHDA injection for 21 days. The number of substantia nigra dopaminergic neurons was detected by immunofluorescence. The expression of Persephin and tyrosine hydroxylase (TH) in the substantia nigra of brain was detected by Western blot. Results Four weeks after injection of Persephin lentivirus, Persephin protein was successfully expressed in dopaminergic neurons (F = 5.451, q = 3.228-4.736, P <0.05). The number of rotations of Persephin lentivirus + 6-OHDA group was significantly lower than that of 6-OHDA group and empty virus + 6-OHDA group (F = 7.551, q = 4.228, 5.336, P <0.05) The number of dopaminergic neurons and the expression of TH were significantly higher than those of 6-OHDA group and empty virus + 6-OHDA group (F = 6.251,5.251, q = 3.228-5.736, P <0.05). Conclusion Persephin has a protective effect on dopaminergic neurons in PD model rats prepared by 6-OHDA.