电针对帕金森病大鼠中脑黑质多巴胺能神经元氧化应激损伤的影响

目的:观察电针对帕金森病(PD)大鼠的中脑黑质超氧化物歧化酶(SOD)活力和谷胱甘肽(GSH)、谷胱甘肽过氧化酶(GSH-Px)、丙二醛(MDA)含量及对中脑黑质酪氨酸羟化酶(TH)表达及多巴胺(DA)能神经细胞凋亡的影响,探讨电针治疗PD的可能机制。方法:雄性Wistar大鼠随机分为正常组(n=10)、模型组(n=11)、电针组(n=11)和西药组(n=11)。皮下注射鱼藤酮溶液制备PD大鼠模型。电针组选"百会""三阴交""太冲"穴电针10min,每天1次,7d为1个疗程,治疗2个疗程;西药组给予美多巴混悬液灌胃(1.67mg/kg)。治疗后行酶法测定右脑中脑黑质SOD、GSH-Px活力,比色定量分析法测定GSH、MDA含量;免疫组化(SP法)染色检测左脑中脑黑质TH,TUNEL法检测DA能神经细胞凋亡。结果:模型组SOD、GSH、GSH-Px均低于正常组(P0.05),模型组MDA高于正常组(P0.05);电针组SOD、GSH、GSH-Px均高于模型组(P0.05),电针组、西药组MDA均低于模型组(P0.05);电针组SOD、GSH-Px均高于西药组(P0.05)。电针组、西药组与模型组相比中脑黑质DA能神经元细胞形态较规则,体积较正常,核仁较明显,神经纤维排列较整齐,结构较密集;西药组改善差于电针组,优于模型组。电针组、西药组与模型组相比TH改善明显,阳性神经元数目增多(P0.05),神经元胞体轮廓及突起清晰;西药组改善差于电针组(P0.05)。电针组、西药组与模型组相比,DA能神经细胞凋亡减少(P0.05)。结论:电针可通过提高PD大鼠中脑黑质SOD、GSH-Px活力和GSH含量,降低MDA含量,增加TH免疫反应阳性神经元数目,抑制DA能神经细胞凋亡来治疗PD。

Effect of Electroacupuncture Intervention on Levels of SOD,GSH,GSH-Px,MDA,and Apoptosis of Dopaminergic Neurons in Substantia Nigra in Rats with Parkinson's Disease

Objective To observe the effect of electroacupuncture(EA)on superoxide(SOD),glutathione peroxidase(GSH-Px)activity,contents of glutathione(GSH)and malondiadehyde(MDA),and expression of tyrosine hydroxylase(TH)and apoptosis of Dopaminergic(DA)neurons in Substantia Nigra of rats with Parkinson's disease(PD).Methods Adult male Wistar rats were randomly divided into normal(10rats),model(11rats),EA(11rats)and medication(11rats)groups.The PD model was established by i.h.of Rotenone(0.8mg/kg)for 28days.EA stimulation(2Hz/80Hz,2mA)was applied at"Baihui"(GV 20),"Sanyinjiao"(SP 6)and"Taichong"(LR 3)acupoints for 10min,once per day for 14times.For rats in the medication group,Madopar suspension fluid(1.67mg/kg)was given by gavage for 14days.Xanthine oxidase method and colorimetric analysis method were used to examine the SOD,GSH-Px activity and contents of GSH and MDA in the Substantia Nigra tissue of the right brain,respectively.Immunohistochemical technique was used to detect the TH positive neurons and TUNEL method was used to examine the apoptosis of DA neurons of the Substantia Nigra in the left brain.Results Following the intervention,the decreased SOD and GSH-Px activity,GSH contents,and the increased MDA content of the Substantia Nigra in PD rats were obviously reversed by EA intervention(P<0.05)but not by medication except MDA content(P>0.05).In comparison with the model group,the decreased TH immunoactivity,and the increased numbers of apoptotic cells of DA neurons were apparently suppressed in both EA and medication groups(P<0.05),but without significant differences between the EA and the medication groups(P>0.05).In addition,HE stain showed that EA intervention could improve PD-induced impairment of Substantia Nigra neurons(mild swelling of neurons with large nucleus and deranged fibers).Conclusion EA intervention can reduce pathological changes of Substantial Nigra in PD rats,which is probably associated with its effects in up-regulating the SOD and GSH-Px activity,GSH contents,and down-regulating MDA level,and reducing the apoptosis of DA neurons of the Substantia Nigra,suggesting an anti-oxidative stress effect of EA therapy.