抑制磷酸腺苷活化蛋白激酶对小鼠脑缺血再灌注后大脑皮质细胞色素C和胱天蛋白酶-3表达及神经细胞凋亡的影响  

Effects of inhibition of adenosine monophosphate-activated protein kinase on expressions of cytochrome c and caspase-3 and neuronal apoptosis in the cerebral cortex after cerebral ischemia-reperfusion injury in mice

作　　者：王展波 李花 郑长亮 陶黎 牟尚东 杨正奇 

机构地区：[1]陕西省汉中市中航工业3201医院神经内科,723000

出　　处：《国际脑血管病杂志》2015年第12期899-902,共4页International Journal of Cerebrovascular Diseases

摘　　要：目的探讨抑制AMP活化的蛋白激酶（AMP-activatedproteinkinase，AMPK）对小鼠脑缺血再灌注后细胞色素C（cytochrome C，CytC）和胱天蛋白酶-3（caspase-3）表达及细胞凋亡的影响。方法36只雄性C57BL／6小鼠随机分为假手术组、缺血再灌注组和AMPK抑制剂组，每组12只。线栓法建立大脑中动脉闭塞模型，AMPK抑制剂组在插入线栓时腹腔注射AMPK抑制剂CompoundC（20mg／kg），假手术组和缺血再灌注组在相同时间点腹腔注射等体积生理盐水。采用免疫组化染色法检测脑缺血再灌注后24h时CytC和胱天蛋白酶-3表达水平，TUNEL法检测细胞凋亡。结果与缺血再灌注组比较，AMPK抑制剂组大脑皮质CytC[（28．86±9．65）个／HP对（58．86±9．65）个／HP；t=7．615，P〈0．001]、胱天蛋白酶-3[（7．16±5．85）个／HP对（14．36±7．85）个／HP；t=2．548，P=0．018]以及TUNEL[（67．14±8．55）个／HP对（95．00±13．51）个／HP；t=6．891，P=0．031]阳性细胞数均显著减少。结论在脑缺血再灌注后抑制AMPK活性可通过下调CytC和胱天蛋白酶表达减少细胞凋亡，发挥神经保护作用。Objective To investigate the effects of inhibition of adenosine monophosphate-activated protein kinase （AMPK） on expressions of cytochrome c （CytC） and caspase-3 and apoptosis in the cerebral cortex after cerebral ischemia-reperfusion injury in mice. Methods Thirty-six male C57BL/6 mice were randomly divided into three groups, a sham operation group, a ischemia-reperfusion group, and a AMPK inhibitor group, 12 in each group. A model of middle cerebral artery occlusion was induced by suture method. The AMPK inhibitor compound C （20 mg/kg） was injected intraperitoneally in the AMPK inhibitor group, the equal volume normal saline was injected intraperitoneally in the sham operation group and the ischemia-reperfusion group when a thread was inserted. Immunohistochemical staining was used to detect the expression levels of CytC and caspase-3 and TUNEL method was used to detect apoptosis at 24 h after ischemia-reperfusion. Results Compared with the ischemia-reperfusion group, the numbers of CytC （28. 86± 9.65/HPvs. 58.86± 9.65/HP; t= 7. 615, P= 0.030） and caspase-3 （7.16± 5.85/HPvs. 14. 36 ±7.85/HP; t =2. 548, P=0. 035）, and TUNEL （67.14 ±8.55/HP vs. 95.00 ± 13.51/HP; t =6. 891, P =0. 030） positive cells in the cerebral cortex were reduced significantly in the AMPK inhibitor group. Conclusion Inhibition of AMPK activity after cerebral ischemia-reperfusion may decrease apoptosis by downregulating the expressions of CytC and caspase-3, and play a neuroprotective effect.

关 键 词：脑缺血 再灌注损伤 AMP-活化蛋白激酶 细胞凋亡 胱天蛋白酶3 细胞色素C类 神经保护药 小鼠 

分 类 号：R743[医药卫生—神经病学与精神病学]