龟羚帕安丸对帕金森病大鼠模型中脑黑质细胞Nestin蛋白,Nestin mRNA及DAT蛋白的影响

目的:观察龟羚帕安丸对帕金森病(PD)大鼠黑质细胞巢蛋白(Nestin),多巴胺转运体(DAT)及Nestin mRNA表达的影响。方法:大鼠黑质致密部注射6-羟基多巴胺(6-OHDA),后大鼠ip阿扑吗啡(APO)诱导旋转建立PD大鼠模型。PD大鼠随机分为模型组,多巴丝肼组,龟羚帕安丸高、中、低剂量组,同时设立假手术组,模型组。假手术组每日ig同等容积的生理盐水;多巴丝肼组ig多巴丝肼混悬液(10 mg·kg~(-1)·d~(-1));龟羚帕安丸高、中、低剂量组ig龟羚帕安丸混悬液(4,2,1 g·kg~(-1)·d~(-1)),连续30 d。免疫组化法检测各组大鼠中脑黑质Nestin,DAT蛋白表达,原位杂交法检测各组大鼠中脑黑质Nestin mRNA的表达。结果:模型组黑质细胞Nestin,DAT蛋白,Nestin mRNA表达均显著低于假手术组(P0.01);各治疗组Nestin蛋白,Nestin mRNA,DAT蛋白表达均显著高于模型组(P0.01)。结论:龟羚帕安丸治疗帕金森病的作用机制可能与上调Nestin,DAT表达,促进神经的修复和再生,增加多巴胺(DA)含量有关。

Effect of Guiling Pa'an Pill on Expression of Nestin, Nestin mRNA and DAT in Substantia Nigra Cells of Rat Models with Parkinson's Disease

Objective: To investigate the effect of Guiling Pa'an pill on the expression of Nestin, Nestin mRNA and dopamine transporter (DAT) in substantia nigra cells of rats with Parkinson's disease (PD). Method: SPF grade rats were stereotaxically injected with 6-hydrox-ydopamine (6-OHDA) and apomorphinum (APO) to induce PD rat models. Rats were randomly divided into model group, madopar group, Guiling Pa'an pill high dose group, middle dose group and low dose group, and at the same time, the sham operation group was also established. Rats in model group and sham operation group daily received the same volume of normal saline. Rats in madopar group received 10 mg · kg^-1 · d^-1 suspension by ig. Rats in Guiling Pa'an pill high dose group, middle dose group and low dose group received 4, 2, 1 g · kg^-1 · d^-1 suspension respectively by ig, all with a course of 30 days. The Nestin and DAT protein levels of the substantia nigra tissue were detected by immunohistochemistry for various groups;Nestin mRNA level in substantia nigra tissues was detected by hybridization in situ method for various groups. Result: Nestin and DAT protein expression and Nestin mRNA expression in substantia nigra tissues of the model group were significantly lower than those in sham operation group (P<0.01). Compared with model group, the Nestin protein, Nestin mRNA and DAT protein expressions in substantia nigra tissues of various treatment groups were significantly increased (P<0.01). Conclusion: Action mechanism of Guiling Pa'an pill for the treatment of Parkinson's disease might be associated with increasing Nestin, DAT expressions, which would promote the repair and regeneration of nerves and increase the content of dopamine.