电针激活Nrf2/HO-1通路抗帕金森病模型小鼠氧化应激的机制研究

Mechanism of Electroacupuncture Against Oxidative Stress in a Mouse Model of Parkinson’s Disease by Activating the Nuclear Factor Erythroid 2-related Factor 2/Heme Oxygenase-1 Pathway

目的观察电针“风府”“太冲”“足三里”穴对帕金森病(Parkinson’s disease,PD)模型小鼠大脑纹状体中酪氨酸羟化酶(tyrosine hydroxylase,TH)、核因子E2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)、血红素加氧酶-1(heme oxygenase-1,HO-1)水平的影响,探讨电针治疗PD的可能机制。方法将30只小鼠按随机数字表法分为对照组、模型组和电针组,每组10只。模型组、电针组小鼠按照30 mg/kg腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)制备PD小鼠模型。电针组小鼠给予“风府”“太冲”“足三里”穴电针干预,每日1次,每次30 min,连续干预12 d。其余两组小鼠不予任何干预。采用爬杆实验和悬挂实验观察3组小鼠行为学改变,旷场实验观察小鼠自主运动总路程,免疫组织化学法检测小鼠大脑纹状体中TH表达水平,实时荧光定量PCR法检测小鼠大脑纹状体中Nrf2、超氧化物歧化酶(super oxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、谷胱甘肽过氧化酶(glutathione peroxidase,GSH-Px)mRNA表达水平;蛋白免疫印迹法检测小鼠大脑纹状体中TH、Nrf2、HO-1蛋白表达水平。结果与模型组比较,电针组小鼠爬杆时间和悬挂时间显著缩短(P<0.05),自主运动总路程显著增加(P<0.05);小鼠大脑纹状体中TH阳性神经纤维较稠密;TH阳性纤维表达水平,Nrf2、SOD、CAT、GSH-Px mRNA表达水平,TH、Nrf2、HO-1蛋白表达水平均显著升高(P<0.05)。结论电针小鼠“风府”“太冲”“足三里”穴能上调大脑纹状体中Nrf2、HO-1表达水平,减轻PD小鼠氧化应激,从而提高纹状体中TH阳性纤维表达,这可能是电针治疗MPTP诱导的PD小鼠行为学障碍的作用机制。

Objective To investigate the effect of electroacupuncture at Fengfu,Taichong,and Zusanli points on the levels of tyrosine hydroxylase(TH),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in the corpus striatum of mice with Parkinson’s disease(PD),as well as the possible mechanism of electroacupuncture in the treatment of PD.Methods A total of 30 mice were divided into control group,model group,and electroacupuncture group using a random number table,with 10 mice in each group.The mice in the model group and the electroacupuncture group were given intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)at a dose of 30 mg/kg to establish a mouse model of PD,and then the mice in the electroacupuncture group were given electroacupuncture at Fengfu,Taichong,and Zusanli points once a day for 30 minutes each time,for 12 consecutive days,while those in the other two groups were not given any treatment.The pole test and the wire hanging test were used to observe behavioral changes;immunohistochemistry was used to measure the expression of TH in the corpus striatum;quantitative real-time PCR was used to measure the mRNA expression levels of Nrf2,superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)in the corpus striatum;Western blotting was used to measure the protein expression levels of TH,Nrf2,and HO-1 in the corpus striatum.Results Compared with the model group,the electroacupuncture group had significant reductions in the time to descend and hanging time(P<0.05),a significant increase in the total distance traveled(P<0.05),dense TH-positive nerve fibers in the corpus striatum of mice,and significant increases in the expression level of TH-positive fibers,the mRNA expression levels of Nrf2,SOD,CAT,and GSH-Px,and the protein expression levels of TH,Nrf2,and HO-1(P<0.05).Conclusion Electroacupuncture at Fengfu,Taichong,and Zusanli points can improve behavioral disturbance induced by MPTP in PD mice by upregulating the expression levels of Nrf2 and HO-1 in the corpus striatum,alleviating oxidative stress in PD mice,and increasing the expression of TH-positive fibers in the corpus striatum.