Qi Ling, Wang Yao, Li Ya-Nan, Guo Lei, Ma Jun
College of Acupuncture-moxibustion and Orthopaedics, Hubei University of Chinese Medicine, Wuhan 430065, China;Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-Hmoxibustion, Wuhan 430065, China.
College of Acupuncture-moxibustion and Orthopaedics, Hubei University of Chinese Medicine, Wuhan 430065, China; Hubei Provincial Collaborative Innovation Center of Preventive Treatment of Diseases by Acupuncture-Hmoxibustion, Wuhan 430065, China.
Zhen Ci Yan Jiu. 2021 Nov 25;46(11):929-34. doi: 10.13702/j.1000-0607.200874.
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.
SD male rats were randomly divided into normal control, PD model and EA groups (=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.
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 (<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 (<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 (<0.05, <0.01).
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.
观察电针“风府”（GV16）、“太冲”（LR3）对鱼藤酮诱导的帕金森病（PD）大鼠中脑黑质酪氨酸羟化酶（TH）、α-突触核蛋白（α-syn）表达及小胶质细胞相关的离子钙接头蛋白1（Iba-1）、Toll样受体4（TLR4）/核因子κB（NF-κB）信号通路的影响，以探讨电针改善PD的作用机制。
将雄性SD大鼠随机分为正常对照组、PD模型组和电针组（每组12只）。采用颈背部皮下注射鱼藤酮（1 mg/kg）制备PD模型。电针双侧GV16、LR3（2 Hz，1 mA），每日1次，连续2周。通过旷场试验对大鼠行为（毛色、反应能力、运动及步态状态）进行评分（0~10分），并检测自主运动状态（自主运动轨迹、总距离、平均速度及8 min内运动持续时间）。采用免疫组织化学染色法检测黑质组织中TH、α-syn的免疫活性，免疫荧光染色法检测Iba-1标记的小胶质细胞数量。采用蛋白质免疫印迹法检测黑质中TLR4、NF-κB P65蛋白表达水平，酶联免疫吸附测定法检测黑质中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）含量。
与正常组比较，模型组大鼠行为学评分、α-syn免疫活性、Iba-1标记的小胶质细胞数量、TLR4及NF-κB P65蛋白表达水平、TNF-α及IL-6含量均显著升高（P<0.01），自主运动总距离、平均速度、8 min内运动持续时间及TH免疫活性显著降低（P<0.01）。电针干预后，与模型组比较，电针组大鼠行为学评分、α-syn免疫活性、Iba-1标记的小胶质细胞数量、TLR4及NF-κB P65蛋白表达水平、TNF-α及IL-6含量升高幅度降低，自主运动总距离、平均速度、8 min内运动持续时间及TH免疫活性降低幅度减小（P<0.05，P<0.01）。
电针可改善PD大鼠行为学表现，其机制可能与下调病理性α-syn异常聚集、TLR4/NF-κB信号通路，抑制小胶质细胞活化及上调中脑黑质TH表达有关。
