针刺调控大鼠脑缺血再灌注损伤Cx43半通道的机制研究

Mechanisms of Cx43 semi-channel in cerebral ischemia-reperfusion injury rats by acupuncture regulation

关键词:  针刺  缺血再灌注  蛋白激酶C  酪蛋白激酶1  蛋白磷酸酶

Key words:  acupuncture  ischemia reperfusion  PKC  CK1  PP2B

摘要:

[目的] 基于连接蛋白43(Cx43)半通道探讨蛋白激酶基因表达在大鼠脑缺血再灌注过程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。采用荧光定量聚合酶链反应(PCR)技术从基因方面检测海马区蛋白激酶C(PKC)、酪蛋白激酶1(CK1)、蛋白磷酸酶(PP2B)的表达。[结果] 与正常组相比,模型组PKC mRNA、CK1 mRNA及PP2B mRNA时间的推移表达量呈增高趋势;与模型组同一时间点比较,非穴位针刺组PKCmRNA表达无显着变化(P>0.05);调神通络针刺组可显着降低其在各时间点表达,与模型组及非穴位针刺组比较,差异具有统计学意义(P<0.05).[结论] 调神通络针刺组可抑制大鼠脑缺血再灌注过程中PKC mRNA、CK1 mRNA及PP2B mRNA表达,通过调节Cx43磷酸化或去磷酸化途径中的关键因子,来调控半通道的开放,在缺血性脑损伤中发挥神经保护作用。

Abstract:

[Objective] To investigate the dynamic changes of protein kinase gene expression in cerebral ischemia and reperfusion rats and the effect of acupuncture intervention based on Cx43 semi-channel. [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 artery in rat focal cerebral ischemia-reperfusion model, the model group and the two acupuncture groups divided into four subgroups: 30 min 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 "top middle" and "top next line"; non-acupoint group were ipsilateral iliac crest on the ribs 10 mm fixed point as acupuncture points; model group did not receive any treatment. Quantitative PCR technology was applied to measure the expression of PKC, CK1and PP2B in the hippocampus. [Results] Compared with the normal group and sham operation group, the expression of PKC mRNA, CK1 mRNA and PP2B mRNA in model group showing a rising trend; compared with the model group at the same time point, and there was no significant change of PKC mRNA expression in the non-acupuncture group (P>0.05). Tiaoshen Tongluo acupuncture can significantly reduce its expression at each time point, compared with the model group and the non-acupuncture group, and the difference was statistically significant (P<0.05). [Conclusion] The Tiaoshen Tongluo acupuncture method may inhibit the expression of PKC mRNA, CK1 mRNA and PP2B mRNA in cerebral ischemia reperfusion rat. Through regulating of key factor phosphorylation of Cx43 phosphate, to control the opening of the semi-channel, playing a neuroprotective role in ischemic brain injury.