针刺研究

2021, v.46(11) 921-928

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电针不同组穴对缺血性脑卒中大鼠神经-血管单元的影响
Effect of electroacupuncture of different acupoint groups on cerebral cortical neurovascular unit and PI3K/Akt signaling in rats with ischemic stroke

李梦醒;付周婷;刘箐;宋宗胜;张慧;唐巍;
LI Meng-xing;FU Zhou-ting;LIU Qing;SONG Zong-sheng;ZHANG Hui;TANG Wei;College of Acupuncture-moxibustion, Anhui University of Chinese Medicine;Department of Rehabilitation, Pizhou Hospital of Traditional Chinese Medicine;Graduate School, Anhui University of Chinese Medicine;

摘要(Abstract):

目的:观察电针对缺血性脑卒中大鼠神经-血管单元的保护作用,探讨脑缺血后电针疗法的作用机制及不同穴位组合的疗效差异。方法:SD大鼠分为假手术组、模型组、电针Ⅰ组和电针Ⅱ组,每组27只。参考Zea Longa线栓法建立右侧大脑中动脉缺血模型。电针Ⅰ组取左侧"曲池""合谷""足三里"及"水沟",电针Ⅱ组取"百会""风府"及左侧"内关""心俞",两电针组均于术后4 h进行电针治疗,每次20 min, 1次/d,共治疗14 d。采用改良神经功能损害评分(mNSS)评价大鼠神经功能改变情况,透射电镜观察缺血区神经元结构,免疫组织化学法检测缺血区CD34阳性细胞表达,实时荧光定量PCR法检测缺血侧大脑运动皮层磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)mRNA表达。结果:与同时点假手术组比较,模型组大鼠mNSS评分升高(P<0.01),神经元结构发生破坏,CD34阳性表达及PI3K、Akt mRNA表达均升高(P<0.01,P<0.05)。与同时点模型组比较,两电针组mNSS评分显著降低(P<0.01),缺血区病理改变均有所好转,CD34阳性细胞表达及PI3K、Akt mRNA表达均明显升高(P<0.01,P<0.05)。与同时点电针Ⅰ组比较,电针Ⅱ组大鼠14 d时mNSS评分降低(P<0.05),7、14 d时CD34阳性表达升高(P<0.01,P<0.05),3、7、14 d时PI3K mRNA表达升高(P<0.05,P<0.01),3、7 d时Akt mRNA表达显著升高(P<0.01)。组内各时间点比较,模型组、电针Ⅰ组、电针Ⅱ组均在14 d时mNSS评分最低(P<0.01);在14 d治疗时程内,模型组、两电针组CD34阳性表达均持续升高(P<0.05,P<0.01),7 d时PI3K、Akt mRNA表达最高(P<0.05,P<0.01)。结论:在电针效应、时间效应及两者的交互作用影响下,脑缺血后电针疗法可改善缺血性脑卒中大鼠的神经功能,其作用机制可能是发挥了对神经-血管单元的保护作用,上调了PI3K/Akt信号通路相关因子表达水平;不同穴位组合对脑缺血神经-血管单元保护作用可能存在差异性,以心与脑的关系指导下的配穴效果更优。
Objective To observe the protective effect of electroacupuncture(EA) on neurovascular unit, neurological function in rats with cerebral ischemia(CI), so as to explore its mechanisms underlying improvement of ischemic cerebral tissue. Methods Male SD rats, SPF grade, were randomly and equally divided into sham operation group, model group, EA group Ⅰ and EA group Ⅱ,27 rats in each group. The CI model was established by occlusion of the middle cerebral artery(MCAO). EA(2 Hz/20 Hz, 0.5 mA) was applied to “Quchi”(LI11), “Hegu”(LI4), “Zusanli”(ST36) and “Shuigou”(GV26) for rats of the EA group Ⅰ, and to “Baihui”(GV26), “Fengfu”(GV16), “Neiguan”(PC26) and “Xinshu”(BL15) for rats of the EA group Ⅱ for 20 min, once a day for 14 days. The modified neurologic severity score(mNSS) was calculated according to the state of locomotor, sensory, and reflex parameters. Transmission electron microscope(TEM) was used to observe the neuronal structure of the ischemic cerebral area. The CD34 positive cells(for microvessels) of the ischemic brain tissue were detected by using immunohistochemistry, and the expression levels of cerebral phosphatidylinositol-3 kinase(PI3 K) and protein kinase B(Akt) mRNAs were detected by quantitative real-time-PCR, respectively. Results Along with the extension of time, the mNSS at 4 h, and 3, 7 and 14 d after CI were apparently decreased, and the number of CD34 positive cells from 3 d to 14 d after CI, and the expression of PI3 K mRNA and Akt mRNA from 3 d to 7 d were significantly increased in the model, EAⅠand EA Ⅱ group(P<0.01, P<0.05). Compared with the sham operation group, the mNSS at 4 h, and 3, 7 and 14 d, and CD34-positive number and PI3 K mRNA and Akt mRNA expression levels at 3, 7 and 14 d were significantly increased in the model group(P<0.01, P<0.05). In comparison with the model group, the mNSS at 3, 7 and 14 d were obviously decreased(P<0.01), and the CD34-positive number and PI3 K and Akt mRNA expression levels at 3, 7 and 14 d considerably increased in both EA group Ⅰ and Ⅱ(P<0.01, P<0.05). The therapeutic effect of EA group Ⅱ was significantly superior to that of EA group Ⅰ in lowering mNSS at 14 d, up-regulating the CD34-positive number at 7 and 14 d, and PI3 K mRNA at 3, 7 and 14 d and Akt mRNA at 3 and 7 d(P<0.05, P<0.01). Results of TEM showed an irregular shape of neurons with nuclear pyknosis, non-uniform chromatin, more organelle loss, swollen mitochondrial Golgi complex and expansion of rough endoplasmic reticulum, being relatively milder in the EA group Ⅰ, particularly in the EA group Ⅱ. Conclusion EA therapy can improve the neurological function in cerebral ischemia rats, which may be related to its effects in protecting the neurovascular unit and up-regulating PI3 K/Akt signal pathway. The effects of EA at GV26, GV16, PC26 and BL15 are better than those of EA at LI11, LI4, ST36 and GV26.

关键词(KeyWords): 缺血性脑卒中;电针;神经功能;神经-血管单元;PI3K/Akt信号通路;穴位组合
Cerebral ischemia;Electroacupuncture;Neurological function;Neurovascular unit;PI3K/Akt signaling pathway;Acupoint combination

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81373711、81874500);; 安徽省自然科学研究青年基金项目(No.1808085QH268);; 安徽省教育厅自然科学基金重点项目(No.KJ2019A0430);; 安徽中医药大学校级探索性科研项目(No.2016ts027)

作者(Author): 李梦醒;付周婷;刘箐;宋宗胜;张慧;唐巍;
LI Meng-xing;FU Zhou-ting;LIU Qing;SONG Zong-sheng;ZHANG Hui;TANG Wei;College of Acupuncture-moxibustion, Anhui University of Chinese Medicine;Department of Rehabilitation, Pizhou Hospital of Traditional Chinese Medicine;Graduate School, Anhui University of Chinese Medicine;

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DOI: 10.13702/j.1000-0607.201051

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