早期电针干预对帕金森病小鼠离子钙接头蛋白-1及肿瘤坏死因子-α表达的影响  

Effects of early electroacupuncture on the expression of Iba-1 and TNF-αin Parkinson’s disease mice

作　　者：马雪 王强 王渊[1,2,3] 袁伟 刘智斌[1,2,3] 乔海法 MA Xue;WANG Qiang;WANG Yuan;YUAN Wei;LIU Zhi-bin;QIAO Hai-fa(College of Acupuncture and Moxibustion,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Shaanxi Key Laboratory of Acupuncture and Herbal Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Shaanxi Provincial Key Laboratory of Traditional Chinese Medicine Encephalopathy,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China)

机构地区：[1]陕西中医药大学针灸推拿学院,陕西咸阳712046 [2]陕西中医药大学陕西省针药结合重点实验室,陕西咸阳712046 [3]陕西中医药大学陕西省中医脑病重点实验室,陕西咸阳712046

出　　处：《针刺研究》2022年第11期993-998,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81873388、81674086);陕西省自然科学基础研究计划(No.2020JM-594);陕西中医药大学经脉-脏腑相关研究创新团队(No.YL-09)。

摘　　要：目的:观察早期电针干预对帕金森病(PD)小鼠离子钙接头蛋白-1(Iba-1)、络氨酸羟化酶(TH)以及肿瘤坏死因子-α(TNF-α)的影响,从神经炎性反应角度探讨电针治疗PD的作用机制。方法:9周龄雄性C57BL/6J小鼠随机分为空白组、模型组、电针组,每组8只。采用长期低剂量皮下注射鱼藤酮制备PD小鼠模型。电针组造模同时予“神庭”“天枢”“曲池”“上巨虚”电针干预,15 min/次,1次/d,连续8周。采用转棒实验、步长实验评估小鼠运动障碍程度;Western blot法和免疫组织化学法检测小鼠中脑黑质区Iba-1、TH蛋白表达水平;Western blot法和免疫荧光染色法检测小鼠结肠组织TNF-α蛋白表达水平。结果:与空白组比较,模型组造模4、6、8周掉落潜伏期缩短(P<0.01),造模5、7周后肢步长减小(P<0.01);与模型组比较,电针组造模6、8周掉落潜伏期延长(P<0.01),造模5、7周后肢步长增大(P<0.01)。与空白组比较,模型组中脑黑质区Iba-1、结肠组织中TNF-α蛋白表达水平升高(P<0.01),中脑黑质区TH蛋白表达水平降低(P<0.01);与模型组比较,电针组中脑黑质区Iba-1、结肠组织中TNF-α蛋白表达水平降低(P<0.01,P<0.05),中脑黑质区TH蛋白表达水平升高(P<0.05,P<0.01)。结论:早期电针干预延缓了鱼藤酮诱发PD小鼠运动障碍发生的时间,减轻了黑质多巴胺能神经元丢失,对PD的退行性改变发挥了良好的神经保护作用,其机制可能与针刺改善肠道炎性反应,抑制小胶质细胞活化,进而减轻神经炎性反应的发生有关。Objective To observe the effect of early electroacupuncture(EA)intervention on ionized calcium binding adapter molecule 1(Iba-1),tyrosine hydroxylase(TH)and tumor necrosis factor-α(TNF-α)in Parkinson’s disease(PD)mice,so as to explore its neuroinflammation mechanism in treating PD.Methods A total of 24 male C57 BL/6 J mice(9 weeks old)were randomly divided into control,model and EA groups,with 8 mice in each group.The PD model was established by long-term low dose subcutaneous injection of rotenone.Started at the same time with modeling,EA(2 Hz/100 Hz,1 mA)was applied to“Shenting”(GV24),bilateral“Tianshu”(LI11),“Quchi”(ST25),and“Shangjuxu”(ST37)for 15 min,once a day for 8 weeks.The motor function was assessed by rotorod test and step length test.The expression levels of Iba-1 and TH proteins in substantia nigra pars compacta(SNpc)was detected by Western blot and immunohistochemistry.The expression level of TNF-αprotein in colon tissue was examined by Western blot and immunofluorescence staining.Results Compared with the control group,the fall latency shortened at 4,6,and 8 weeks after modeling(P<0.01)and the step length of hind limbs shortened at 5 and 7 weeks after modeling(P<0.01),the expression levels of Iba-1 in SNpc and TNF-αin colon tissue were increased(P<0.01),and the expression level of TH in SNpc was decreased(P<0.01)in the model group.Compared with the model group,the fall latency prolonged at 6 and 8 weeks after modeling(P<0.01)and the step length of hind limbs prolonged at 5 and 7 weeks after modeling(P<0.01),the expression levels of Iba-1 in SNpc and TNF-αin colon tissue were decreased(P<0.01,P<0.05),and the expression level of TH in SNpc was increased(P<0.05,P<0.01)in the EA group.Conclusion Early EA intervention can delay the occurring time of motor disfunction,alleviated the loss of substantia nigra dopaminergic neurons,and exerted a good neuroprotective effect on the degenerative changes in rotenone-induced PD mice,which may be related to its effects in alleviating the int

关 键 词：帕金森病 神经炎性反应 外周炎性反应 脑肠轴 电针 

分 类 号：R245.97[医药卫生—针灸推拿学]