针刺研究

2021, v.46(11) 929-934+972

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电针对鱼藤酮诱导的帕金森病大鼠神经炎性反应及Toll样受体4/核因子-κB信号通路的影响
Electroacupuncture improves behavioral activities by suppressing neuroinflammation and TLR4/NF-κB signaling in substantia nigra of midbrain in Parkinson’s disease rats

祁羚;汪瑶;李亚楠;郭磊;马骏;
QI Ling;WANG Yao;LI Ya-nan;GUO Lei;MA Jun;College of Acupuncture-moxibustion and Orthopaedics,Hubei University of Chinese Medicine;Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-Hmoxibustion;

摘要(Abstract):

目的:观察电针"风府""太冲"穴对鱼藤酮诱导的帕金森病(Parkinson’s disease, PD)大鼠中脑黑质酪氨酸羟化酶(TH)、α-突触核蛋白(α-syn),小胶质细胞(MG)以及Toll样受体4(TLR4)/核因子(NF)-κB信号通路相关蛋白的影响,探讨电针治疗PD的可能作用机制。方法:SD大鼠随机分为正常组、模型组、电针组,每组12只。采取颈背部皮下注射鱼藤酮(1 mg/kg)制备PD大鼠模型。电针组电针"风府"和"太冲",30 min/次,每日1次,连续治疗14 d。采用行为学评分和敞箱实验检测各组大鼠行为学改变,免疫组织化学法检测大鼠中脑黑质TH、α-syn的表达水平,免疫荧光标记法检测中脑黑质激活的MG数量,蛋白免疫印迹法检测大鼠中脑黑质TLR4、NF-κB P65,酶联免疫吸附法检测中脑黑质肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)等炎性因子水平。结果:与正常组比较,模型组大鼠行为学评分升高(P<0.01),自主运动总路程、平均速度、运动持续时间减少(P<0.01);中脑黑质TH表达减少(P<0.01),α-syn表达、激活的MG数量、TLR4及NF-κB P65蛋白表达、TNF-α及IL-6含量增加(P<0.01)。与模型组比较,电针组行为学评分降低,自主运动总路程、平均速度、运动持续时间增加(P<0.01);中脑黑质TH表达增多(P<0.01),α-syn表达、激活的MG数量、TLR4及NF-κB P65蛋白表达、TNF-α及IL-6含量减少(P<0.01,P<0.05)。结论:电针能有效改善PD大鼠运动迟缓等行为学表现,减少α-syn的异常聚集,提高中脑黑质TH表达水平,其相关机制可能与抑制神经炎性反应及TLR4/NF-κB信号通路相关蛋白有关。
Objective To observe the effect of electroacupuncture(EA) at “Fengfu”(GV16) and “Taichong”(LR3) on the expression of tyrosine hydroxylase(TH), α-synuclein(α-syn) and microglial-related microglial(MG), Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) signaling pathway in the substantia nigra(SN) of midbrain in Rotenone-induced Parkinson's disease(PD) rats, so as to explore its mechanisms underlying improvement of PD. Methods SD male rats were randomly divided into normal control, PD model and EA groups(n=12 in each group). The PD model was established by subcutaneous injection of rotenone(1 mg/kg) at the back of neck. EA(2 Hz, 1 mA) was applied to bilateral GV16 and LR3, once daily for 2 weeks. The rats' behavior(hair color, reaction capacity, locomotion and gait state)scores(0-10 points) were given and the autonomic movement state(trajectory of autonomous motion, total distance, average speed and duration of motion in 8 min) was detected by using open field tests. The immunoactivity of TH and α-syn in the SN tissue were determined by using immunohistochemistry staining, and the number of Iba-1-labelled microglia(MG) was detected by using immunofluorescence staining. The expression levels of TLR4 and NF-κB P65 proteins in the SN were detected by Western blot, and the contents of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in the SN were determined by using enzyme-linked immunosorbent assay. Results In comparison with the normal group, the behavioral score, α-syn immunoactivity, number of Iba-1 labelled microglia, expression of TLR4 and NF-κB P65 proteins and the contents of TNF-α and IL-6 in the SN were significantly increased(P<0.01), whereas the total distance, average speed, duration of motion of the autonomic movement in 8 min, and the TH immunoactivity were remarkably decreased in the model group(P<0.01). After EA intervention, compared with the model group, the increase of the behavioral score, α-syn immunoactivity, number of Iba-1-labelled microglia, expression of TLR4 and NF-κB P65 proteins, the contents of TNF-α and IL-6 in the SN, and the decrease of the total distance, average speed, duration of motion of the autonomic movement in 8 min, and the TH immunoactivity were reversed(P<0.05, P<0.01). Conclusion EA can improve the behavioral manifestations of PD rats, which may be associated with its functions in down-regulating the abnormal accumulation of pathological α-syn, TLR4/NF-κB signaling, inhibiting activities of microglia and in up-regulating the expression of TH in the SN of midbrain.

关键词(KeyWords): 帕金森病;电针;Toll样受体4;核因子κB;小胶质细胞;炎性因子
Parkinson's Disease;Electroacupuncture;Toll-like receptor 4;Nuclear factor kappa B;Microglia;Inflammatory factor

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81473788);; 国家重点研发计划项目(No.2020YFC0845800)

作者(Author): 祁羚;汪瑶;李亚楠;郭磊;马骏;
QI Ling;WANG Yao;LI Ya-nan;GUO Lei;MA Jun;College of Acupuncture-moxibustion and Orthopaedics,Hubei University of Chinese Medicine;Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-Hmoxibustion;

Email:

DOI: 10.13702/j.1000-0607.200874

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