大鼠脑缺血再灌注损伤Cx43半通道的开放以及针刺干预的影响  

Opening of Cx43 Semi- Channel after Cerebral Ischemia Reperfusion Injury and Its Effect by Acupuncture Intervention

作　　者：刘坤[1] 张连城[1] 郭家奎[1] 杨宝旺[1] 肖震心[1] 潘艳颖[1] 马妍[2] 郭富斌[3] 王占奎[1] 

机构地区：[1]天津中医药大学第二附属医院,天津300150 [2]天津中医药大学,天津300193 [3]长春中医药大学附属医院,吉林长春130021

出　　处：《辽宁中医杂志》2015年第8期1535-1538,I0001,I0002,共6页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81102640);天津市应用基础与前沿技术研究计划重点项目(14JCZDJC36800)

摘　　要：目的:探讨局灶脑缺血再灌注大鼠缝隙连接蛋白Cx43半通道的开放在缺血性脑损害中的动态变化规律以及针刺干预的影响。方法:根据随机数字表将Wistar大鼠随机分为正常组、模型组、非穴位针刺组及针刺组。采用改良的线栓法制备大鼠左侧大脑中动脉局灶性脑缺血再灌注模型,模型组及两个针刺组内分设4个亚组:缺血再灌注30 min组、缺血再灌注60 min组、缺血再灌注180 min组、缺血再灌注360 min组,每组10只大鼠。穴位组电针刺激大鼠的"顶中线""顶旁线";非穴位组取患侧肋下髂嵴上10 mm的固定点作为针刺点;模型组不予任何处理。采用激光共聚焦显微技术和Ca2+特异性荧光标记方法,结合Cx43半通道阻断剂观察半通道在缺血再灌注过程中对神经元细胞内钙稳态的影响。结果:大鼠脑缺血再灌注30 min时,脑组织海马CA1区神经元细胞内[Ca2+]i开始升高;到再灌注360 min时达到高峰;调神通络针刺组对脑缺血再灌注大鼠各时程的[Ca2+]i都有降低作用,与模型组比较,差异具有统计学意义(P<0.05),非穴位针刺组与模型组同一时间点比较,[Ca2+]i无显著变化(P>0.05)。结论:阻断半通道的开放可以有效降低缺血再灌初期的钙超载程度;"调神通络"针法组对脑缺血再灌注大鼠各时程的[Ca2+]i都有降低作用,在缺血性脑损伤中发挥神经保护作用。Objective ：To investigate the dynamic changes of semi -channel in focal cerebral ischemi~ reperfusion rats and its effect by acupuncture intervention. Methods ： According to the random number table, Wistar rats were randomly divided into normal group, model group, the non - acupuncture group and the acupuncture group. Using a modified intraluminal left middle cerebral ar- tery in rat focal cerebral ischemia - reperfusion model, the model group and the two acupuncture groups were divided into four subgroups ;30 rain ischemia- reperfusion group, ischemia -reperfusion 60 min group, ischemia -reperfusion group 180 min and 360 min ischemia - reperfusion groups. Electrical stimulation of acupuncture group were middle line of Vertex and lateral line bf vertex;non -acupoint group chose ipsilateral iliac crest on the ribs 10 mm fixed point as acupuncture points ;model group did not receive any treatment. Confocal laser scanning microscopy method and Ca2＋ specific fluorescent labeling method were applied to observe the process of neuronal intraceUular calcium homeostasis in the process of ischemia ＇ reperfusion. Results： [ Ca2＋ ] i in hippocampal CA1 neurons began to rise 30 min after the cerebral ischemia and reperfusion and reached to peak 360 min after ischemia - reperfusion. Compared with the model group, Tiaoshen Tongluo acupuncture had a lowering effect of [ Ca2＋ ] i at the same time point and the difference was statistically significant （ P 〉 0.05 ） while the [ Ca2＋ ] i in the non - acupuncture group showed no significant changes （P 〉0. 05 ）. Conclusion：The semi - open channel blockers can reduce calcium overload level of isehemia - reperfusion early. Tiaoshen Tongluo acupuncture has a lowering＇ effect of [ Ca2＋] i in cerebral ischemia and reperfusion rats,playing a neuroprotective role in ischemic brain injury.

关 键 词：针刺 缺血再灌注 半通道 钙超载 

分 类 号：R246.6[医药卫生—针灸推拿学]