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黄慧源,易丽贞,黄麟荇,陈瑞雪,绽晟,岳增辉.电针调控小胶质细胞极化抑制脑卒中肢体痉挛大鼠脑皮质炎症反应的研究[J].湖南中医药大学学报英文版,2023,43(12):2256-2263.[Click to copy
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| 电针调控小胶质细胞极化抑制脑卒中肢体痉挛大鼠脑皮质炎症反应的研究 |
| 黄慧源,易丽贞,黄麟荇,陈瑞雪,绽晟,岳增辉 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察电针阳陵泉、曲池对脑卒中肢体痉挛（post-stroke spasticity，PSS）大鼠脑皮质中小胶质细胞极化及炎症因子的影响，探讨电针缓解PSS的可能机制。方法 采用改良大脑中动脉闭塞（middle cerebral artery occlusion，MCAO）线栓法+内囊注射N-甲基-D-天冬氨酸（N-Methyl-D-aspartic acid，NMDA）受体法制备PSS模型。随机抽取10只大鼠作为假手术组，剩余大鼠造模成功后选20只再随机分为模型组和电针组，每组10只。电针组干预患侧阳陵泉、曲池，每天治疗1次，每次30 min，连续治疗7 d；假手术组和模型组仅进行同条件固定，不作其他干预。通过神经功能缺损评分、电生理描记法评估电针对PSS大鼠的治疗效果；免疫荧光染色法观察大鼠缺血侧脑皮质小胶质细胞标志物离子钙结合衔接分子-1 （ionized calcium binding adapter molecule-1，Iba-1）、M1型标志物白细胞分化抗原16（cluster of differentiation 16，CD16）、M2型标志物白细胞分化抗原206（cluster of differentiation206，CD206）的阳性表达；ELISA法和qPCR法检测大鼠缺血侧脑皮质中白细胞介素-6（interleukin-6，IL-6）、白细胞介素-1β（interleukin-1β，IL-1β）、转化生长因子-β（transforming growth factor-β，TGF-β）的含量和mRNA表达情况。结果 与假手术组比较，模型组大鼠神经功能缺损评分明显升高（P<0.01），间接肌张力明显下降（P<0.01），Iba-1和CD16阳性荧光强度表达显著增加（P<0.01），CD206阳性荧光强度表达显著下降（P<0.01），IL-6、IL-1β含量及mRNA表达明显上升（P<0.01），TGF-β含量和mRNA表达明显下降（P<0.01）。治疗后与模型组比较，电针组大鼠神经功能缺损评分降低（P<0.05），间接肌张力明显上升（P<0.01），Iba-1阳性荧光强度表达减少（P<0.05），CD16阳性荧光强度表达明显下降（P<0.01），CD206阳性荧光强度表达显著上升（P<0.01），IL-6含量及mRNA的表达下降（P<0.05），IL-1β含量及mRNA表达明显下降（P<0.01），TGF-β含量和mRNA表达明显上升（P<0.01）。结论 电针阳陵泉、曲池能够有效改善PSS大鼠神经功能损伤、降低肌张力、缓解痉挛症状，其机制可能与抑制小胶质细胞激活、调控小胶质细胞由M1向M2极化、缓解神经炎症有关。 |
| 关键词: 脑卒中 肢体痉挛 电针 阳陵泉 曲池 小胶质细胞 炎症反应 |
| DOI：10.3969/j.issn.1674-070X.2023.12.016 |
| Received:September 11, 2023 |
| 基金项目:国家自然科学基金面上项目（81673886）；湖南省自然科学基金面上项目（2023JJ30462）；湖南省研究生科研创新项目（CX20220797）；湖南中医药大学研究生科研创新项目（2022CX19，2022CX110，2023CX78）。 |
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| Electroacupuncture regulates microglia polarization to inhibit inflammatory response in cerebral cortex of rats with post-stroke spasticity |
| HUANG Huiyuan,YI Lizhen,HUANG Linxing,CHEN Ruixue,ZHAN Sheng,YUE Zenghui |
| (School of Acupuncture-moxibustion, Tuina, and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effects of electroacupuncture at "Yanglingquan" (GB34) and "Quchi" (LI11) on microglia polarization and inflammatory factors in the cerebral cortex of rats with post-stroke spasticity(PSS), so as to explore the possible mechanism of electroacupuncture in alleviating PSS.Methods A PSS model was prepared by the modified middle cerebral artery occlusion(MCAO) suture method + internal capsule injection of N-Methyl-D-aspartic acid(NMDA) receptor. Ten rats were randomly selected as the sham-operated group. After successful modeling, 20 rats were selected and randomly subdivided into model group and electroacupuncture group, with 10 rats in each group. The electroacupuncture group was intervened with electroacupuncture at "Yanglingquan" (GB34) and "Quchi" (LI11) on the affected side, once a day, 30 min each time, for 7 consecutive days, while the sham-operated group and the model group were only fixed under the same conditions without any other intervention. The therapeutic effects of electroacupuncture on PSS rats were evaluated through neurological deficit score and electrophysiological tracing; immunofluorescence staining was used to observe the positive expressions of microglia marker ionized calcium binding adapter molecule-1(Iba-1), M1 phenotype marker cluster of differentiation 16(CD16), and M2 phenotype marker cluster of differentiation 206(CD206) in ischemic cerebral cortex of rats; ELISA and qPCR were used to determine the content and m RNA expressions of interleukin-6(IL-6), interleukin-1β(IL-1β), and transforming growth factor-β(TGF-β) in the ischemic cerebral cortex of rats.Results Compared with the sham-operated group, the neurological deficit score of the model group significantly increased(P<0.01), the indirect muscle tone markedly decreased(P<0.01), the expressions of Iba-1 and CD16 positive fluorescence intensity significantly elevated(P<0.01), the expression of CD206 positive fluorescence intensity significantly declined(P<0.01), the content and mRNA expressions of IL-6 and IL-1βwere significantly higher(P<0.01), and the content and mRNA expression of TGF-βwere obviously lower(P<0.01). After treatment, compared with the model group, the neurological deficit score of electroacupuncture group decreased(P<0.05), the indirect muscle tone significantly increased(P<0.01), Iba-1 positive fluorescence intensity expression declined(P<0.05), CD16 positive fluorescence intensity expression was obviously lower(P<0.01), CD206 positive fluorescence intensity expression markedly elevated(P<0.01), the content and mRNA expressions of IL-6 and IL-1β decreased(P<0.05), TGF-β content and mRNA expression increased obviously(P<0.01).Conclusion Electroacupuncture at "Yanglingquan" (GB34) and "Quchi" (LI11) can effectively reduce neurological impairment, decrease muscle tone, and alleviate spastic symptoms in PSS rats. The mechanism may be related to inhibiting the activation of microglia, regulating microglia polarization from M1 to M2, and alleviating neuroinflammation. |
| Key words: stroke limb spasticity electroacupuncture "Yanglingquan" (GB34) "Quchi" (LI11) microglia inflammatory response |
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