抑制AMPK活性对小鼠脑缺血/再灌注损伤后星形胶质细胞形态及功能的影响  

The Effectsof Inhibition of AMPK Activity on the Morphology and Function of Astrocytesafter Cerebral Ischemia/reperfusion Injury in Mice

作　　者：沙晶[1] 党辉[1] 艾山江[1] 玛依努尔[1] 朱沂[1] 李红燕[1] 

机构地区：[1]新疆自治区人民医院神经内科,乌鲁木齐830001

出　　处：《新疆医学》2016年第5期512-516,507,共6页Xinjiang Medical Journal

基　　金：国家自然科学基金(81060104)

摘　　要：目的探讨磷酸腺苷活化蛋白激酶(AMP-actives protein kinase,AMPK)抑制剂Compound C对脑缺血/再灌注小鼠星形胶质细胞形态及其功能的影响,研究抑制AMPK活性的脑保护作用机制。方法采用线栓法建立昆明小鼠右侧大脑中动脉闭塞再灌注模型(middle cerebral artery occlusion,MCAO),应用Longa神经功能评分法测定各组小鼠神经功能缺损评分,TTC染色法观察各组小鼠脑梗死体积大小;采用免疫组化法观察星形胶质细胞特异性标记物胶质纤维酸性蛋白(Glia fiber acid protein,GFAP)的表达;采用R T-PCR法观察活化星形胶质细胞连接蛋白43(Connexin 43,Cx43)及兴奋性氨基酸2(Excitatory amino acid transporters-2,EAAT2)的表达。结果小鼠脑缺血/再灌注24 h后,与盐水对照组相比,药物干预组组可以显著减少脑梗死的体积(P<0.01),改善神经功能评分(P<0.05);减少星形胶质细胞的活化(P<0.05);可上调小鼠梗死侧脑组织Cx43的表达(P<0.05);而EAAT2表达具有上升趋势,但无统计学差异(P=0.235)。结论应用Compound C抑制AMPK活性对小鼠局灶性脑缺血再灌注损伤有一定的神经保护作用,其作用机制可能与抑制星形胶质细胞过度活化、增加Cx43及EAAT2表达相关。Objective To observe the effects of Compound C ,an AMPK inhibition, on the morphology and function of astrocytes in the ischemia/reperfusion mice brain, and to study the neuroprotection mechanism about inhibiting AMPK activity. Methods Male Kunming mice were subjected to right middle cerebral artery occlusion （MCAO）. Infarct volumes and behavioral outcome were determined to evaluate the protection of Compound C injection on cerebral ischemic/reperfusion injury. The expression of astrecyte-specific glial fibrillary acidic protein （GFAP） were detected by immunohistochemistry to observe the morphology of active astrocytes. Glutamate transporter-1 （GLT-1） and connexin 43 （Cx43） were detected by RT-PCR. Results After being cerebral ischemia/reperfusion for 24 h, infarct volume and the neurological function score in experimental group were decreased significantly compared with the saline group （P〈 0.01, P 〈0.05, respectively）; And GFAP-labeled cells were also decreased significantly compared with the saline group （P 〈0.05）; Compound C could up-regnlate the expression of Cx43 mRNA in infarct side （P 〈0.05）; In the experimental group after being given Compound C, it showed an upward trend of EAAT2 expression, but there were no statistically significant difference compared with the control group（P=0.235）. Conclusion The present study provides an evidence in vivo that Compound C, an AMPK inhibition, could have neuroprotection effects on the ischemia/reperfusion mice brain. The mechanism is related to inhibiting GFAP-labeled cells, up-regulating Cx43and EAAT2 expression.

关 键 词：AMPK 脑缺血再灌注损伤模型 星形胶质细胞 神经保护作用 

分 类 号：R743.3[医药卫生—神经病学与精神病学]