应用蛋白芯片技术筛选针刺对脑缺血再灌注损伤大鼠的凋亡相关蛋白

摘要

目的应用蛋白芯片技术筛选针刺对缺血再灌注损伤( CIRI)大鼠脑组织凋亡相关蛋白。方法SD健康大鼠40只随机分为4组：假手术组、模型组、针刺对照点组(均取穴左侧旁开0.3 cm的非经非穴点)、针刺穴位组(针刺大椎、百会、人中)，每组10只，各组大鼠6次处理后取左侧海马脑组织，采用720磷酸化抗体蛋白芯片检测处理后的脑组织细胞信号蛋白磷酸化的变化，然后筛选各组上调、下调显著(磷酸化水平变化上调≥1.5倍，下调≤0.67倍)且与凋亡相关的蛋白。结果与假手术组比较，模型组上调≥1.5倍与凋亡相关的蛋白8种，下调≤0.67倍的蛋白4种；与模型组比较，对照点组上调≥1.5倍与凋亡相关的蛋白5个，下调≤0.67倍的蛋白8个；穴位组上调≥1.5倍与凋亡相关的蛋白2个，下调≤0.67倍的蛋白11个，其中下调模型组中上调的蛋白3种。结论针刺大椎、百会、人中(穴)对CIRI大鼠通过调整多个凋亡相关的蛋白表达变化抑制凋亡实现脑保护作用。%Objective To screen the apoptosis-related protein affected by acupuncture in cerebral ischemia reperfu-sion injury ( CIRI) rats using the technology of protein chip. Methods 40 healthy SD rats were randomly divided into four groups,sham operation group,model group,acupuncture points control group( located 0.3 cm from the left side of acu-points,which are not meridian and acupoints),acupuncture group (Dazhui,Baihui,Renzhong are acupunctured),with 10 rats in each group. We extracted ischemic brain tissue after 6 times treatments,and detected the phosphorylations of insulin signaling proteins after acupuncture treatment by using protein chip technology with 720 phosphorylation antibody,and then screened the apoptosis-related proteins which were up-regulated or down-regulated significantly ( The increasing of the phos-phorylation level ≥1. 5 times,decreasing ≤0. 67 times ). Results (1) Compared with the sham group,there were 8 kinds of apoptosis-related proteins in model group which were increased for ≥1. 5 times,and 4 kinds of apoptosis-related proteins which were decreased for≤0. 67 times;(2) Compared with the model group,there were 5 kinds of apoptosis-relat-ed proteins in the control point group which were increased for≥1. 5 times,and 8 kinds of apoptosis-related proteins which were decreased for ≤ 0. 67 times;there were 2 kinds of apoptosis-related proteins in acupuncture group which were in-creased for ≥1. 5 times,and 11 kinds of apoptosis-related proteins which were decreased for ≤ 0. 67 times,there were 3 kinds which were increased in the model group were decreased. Conclusion Brain protection can be realized by inhibi-ting apoptosis with the regulation of several apoptosis-related proteins after acupuncture treatment on Dazhui, Baihui, Re-nzhong ( acupoints) in CIRI rats.