国际麻醉学与复苏杂志   2015, Issue (6): 8-8
    
丝裂原和应激激活蛋白激酶1及cAMP反应元件结合蛋白参与小剂量氯胺酮降低小鼠缺血性脑损伤
舒洛娃1()
1.首都医科大学附属北京同仁医院
Mitogen?蛳and stress?蛳activated protein kinase 1 and cAMP response element binding protein is involved in low dose ketamine-induced neuroprotection against cerebral ischemic injuries of mice
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摘要：

目的　探讨丝裂原和应激激活蛋白激酶1（mitogen-and stress-activated protein kinase 1， MSK1）及cAMP反应元件结合蛋白（cAMP response element binding protein， CREB）在氯胺酮保护小鼠缺血脑组织中的作用。　方法　80只健康成年雄性BALB/c小鼠按完全随机法分为5组［每组16只，其中包括行为学检测、氯化三苯基四氮唑（triphenyltetrazolium chloride， TTC）染色10只，Western blot 检测6只］：假手术组，大脑中动脉阻塞（middle cerebral artery occlusion， MCAO）+生理盐水组，MCAO+25、50、100 mg/kg氯胺酮组。利用小鼠MCAO模型，结合神经行为学评分、TTC染色和Western blot等技术，检测小鼠脑皮质不同缺血区域内MSK1和CREB磷酸化水平和蛋白总量的表达，观察不同剂量氯胺酮对小鼠脑缺血损伤的影响。　结果　与MCAO+生理盐水组比较，MCAO+25 mg/kg氯胺酮组小鼠脑缺血后神经行为学表现明显改善［（8.2±0.4） 分比（6.7±0.3） 分］，脑缺血皮质梗死体积减少［（28.3±2.0）%比（21.0±2.2）%］，水肿率降低［（10.6±0.4）%比（6.9±0.9）%］，缺血半影区内MSK1［（52.3±7.7）%比（81.1±6.9）%］和CREB［（56.6±5.9）%比（78.6±7.1）%］磷酸化水平明显增加（P<0.05）；而MSK1和CREB总蛋白表达量在缺血核心区和半影区内均无明显改变。MCAO+50、100 mg/kg氯胺酮组小鼠行为学表现，缺血皮质梗死体积，水肿率，及缺血半影区MSK1、CREB磷酸化水平无明显改变。　结论　小剂量氯胺酮降低小鼠缺血性脑损伤可能与缺血皮质半影区MSK1及其底物CREB的磷酸化水平增高有关。
关键词： 氯胺酮；　大脑中动脉阻塞；　脑损伤；　丝裂原和应激激活蛋白激酶1；　cAMP反应
Abstract:

Ｏｂｊｅｃｔｉｖｅ　To explore the role of mitogen-and stress-activated protein kinase 1（MSK1） and cAMP response element binding protein（CREB） in ketamine-induced neuroprotection against cerebral ischemic injuries.　Methods　Eighty healthy male BALB/c mice were randomly divided into 5 groups as follows[ n=16, neurological deficits test， triphenyltetrazolium chloride（TTC） staining 10 mice per group， western blot test 6 mice per group]： sham-operation， middle cerebral artery occlusion（MCAO）+saline， MCAO+25， 50， 100 mg/kg ketamine. Using MCAO-induced focal cerebral ischemia mouse models and the techniques of triphenyltetrazolium chloride（TTC） staining， neurological deficits evaluation and Western blot we observed the neurological scores， brain infarct volume， the phosphorylation and protein expression level of MSK1 and CREB in regions of ischemic cortex to study the effects of low doses ketamine on mouse cerebral ischemic injuries.　Results　MCAO+25 mg/kg ketamine attenuated MCAO?蛳induced neurological deficits ［（8.2±0.4）% vs （6.7±0.3）%］， decreased brain infarct volume［（28.3±2.0）% vs （21.0±2.2）%］ and edema ratio［（10.6±0.4）% vs （6.9±0.9）%］， 25 mg/kg ketamine also significantly enhanced the phosphorylation levels of MSK1 ［（52.3±7.7）% vs （81.1±6.9）%］ and CREB［（56.6±5.9）% vs （78.6±7.1）%］ in penumbra of MCAO-induced ischemic cortex. However， there was no significant changes of total MSK1， CREB protein expressions in the ischemic regions of ketamine-treated MCAO mice. There was no significant change of neurological deficits， infarct volume， edema ratio， and phosphorylation level of MSK1， CREB in penumbra of cortex in MCAO+50， 100 mg/kg ketamine group.　Conclusions　The increased phosphorylation levels of MSK1 and CREB in ischemic penumbra were involved in low dose ketamine-induced neuroprotection against mouse cerebral ischemic injuries.
Key words: Ketamine；　Middle cerebral artery occlusion；　Cerebral ischemic injury；　Mitogen-and stress-activated protein kinase 1；　cAMP response element binding protein