摘要
目的:通过建立百草枯(Paraquat,PQ)诱导的帕金森病(Parkinson’s disease,PD)小鼠模型,观察绿茶多酚及灸百会穴对百草枯诱导PD的保护作用,探讨其机制。方法:选用8周龄雄性C57BL/6小鼠65只随机分为阴性对照组、百草枯组、绿茶多酚组、灸百会穴组、及绿茶多酚+灸百会穴组,除阴性对照组外,每组均按10mg/kg给予PQ,用滴管经口腔染毒,生理盐水作为阴性对照,绿茶多酚组、灸百会穴组、及绿茶多酚+灸百会穴组给予保护措施。采用爬杆实验观察小鼠行为学改变,是否出现类帕金森病症状;8周后断头取脑,采用HE染色观察中脑黑质细胞变化,应用高效液相紫外荧光法测定脑多巴胺(DA)含量。结果:与阴性对照组相比,百草枯组出现震颤、竖毛、前腿抬高、竖尾、动作缓慢和减少等类帕金森症状明显,各实验组小鼠出现上述症状时间无明显差别,阴性对照组和给予保护措施组的小鼠行为改变与百草枯组相比均有统计学意义(P <0.01),但爬竿试验时间变化无统计学意义(P >0.05),病理显示:百草枯组与阴性对照及保护剂实验组比较黑质区多巴胺能神经元细胞数量减少明显,密度降低,但各保护剂组之间未见明显差别。高效液相结果示:与对照组相比,脑内DA的含量有所减少,且百草枯组减少明显,但差别无统计学意义(P >0.05)。结论:百草枯可诱导C57小鼠出现类帕金森病症状,使中脑黑质部多巴胺能神经元数目减少,其发病呈慢性进行性过程,绿茶多酚及灸百会穴对百草枯诱导的PD小鼠有一定的保护作用,其机制有待于进一步探讨。
Objective: To investigate the effect of green tea polyphenols and acupuncture on the behavior and the dopamine level of paraquat induced Parkinson’s disease mice model, and to explore the protection and its mechamism in those PD mice model. Methods: Eight-week-old male C57 mice were divided into 5 groups randomly: negative control, paraquat, green tea polyphenols, acupuncture,green tea polyphenols and acupuncture, dose of 10mg/kg paraquat exposed to all the groups except negative control. And with the exception of the negative control and paraquat group, the others treated with protective medium. The changes of behavior were observed at different periods with pole-test, to determine whether those mice have PD symptom. After 8 weeks, the mice were killed with decapitation, the changes of neurons of the substantia nigra in midbrain were observed with HE staining, And the level of dopamine in brain was measured with HPLC-UV. Results: The symptoms like tremor, piloerection, raising,ante-limb,slow-moving,and aberrant behavior in paraquat group were more obviously than others. Compared with control, the score of praxiology in paraquat group had statistical significance(P<0.05); but the appearance time of such symptoms and pole-test time had no statistical significance(P>0.05). Pathological changes showed that, dopaminergic neurons decreased and density losed obviously in paraquat group compared with control, but obvious differences were not found among protective additive groups. Compared with control group, the level of DA decreased and obviously in paraquat with HPLC-UV, but the difference had no statistical significance (P>0.05). Conclusion: The tests prompt that PQ can make C57 mice appear Parkinsonism symptom and decrease dopaminergic neurons in substantial nigra, and the process of the disease is chronic. Green tea polyphenols and acupuncture really improve the behavior of paraquat injury C57 mice, but the mechanism needs further investigation.
引文
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