ERK和CREB磷酸化在硫氢化钠对大鼠脑缺血再灌注神经保护的机制探讨  

Roles of ERK and CREB phosphorylation in sodium hydrosulfide-induced neuroprotection after cerebral ischemia-reperfusion in rats

作　　者：殷俊[1] 李艳冰[2] 沈琴[1] 杨晓苏[1] 

机构地区：[1]中南大学湘雅医院神经内科,湖南省长沙市410008 [2]中南大学湘雅医院检验科,湖南省长沙市410008

出　　处：《国际神经病学神经外科学杂志》2015年第3期223-229,共7页Journal of International Neurology and Neurosurgery

基　　金：2015年"步长杯"脑血管病科学研究基金项目(BC2015-02)

摘　　要：目的探讨磷酸化细胞外调节蛋白激酶(p-ERK)和磷酸化cAMP反应元件结合蛋白(p-CREB)在外源性硫化氢(NaHS)干预下对大鼠脑缺血再灌注的神经保护作用及机制。方法将180只雄性SD大鼠随机分为3组:假手术组、模型组和NaHS干预组(100μmol/L),每组60只。线栓法制备大鼠大脑中动脉栓塞模型,2h后再灌注,灌注前20min给予生理盐水或NaHS腹腔注射,术后6h、1d、2d、5d、7d后行TTC染色观察脑梗死体积,尼氏染色检测神经元表达,WesternBlot检测海马p-ERK1/2、p-CREB表达,免疫组化检测Bcl-2表达。结果干预组脑梗死体积和神经元凋亡相对于模型组显著减少(P<0.05)。与模型组相比,干预组各时间点p-ERK1/2、p-CREB蛋白表达均升高(P<0.05),两者之间呈正相关;干预组Bcl-2相对于模型组升高(P<0.05)。结论大鼠脑缺血再灌注损伤后NaHS可能通过诱导ERK1/2和CREB蛋白磷酸化,激活下游Bcl-2蛋白表达,起到抗凋亡作用,减少脑梗死体积,发挥神经保护作用。Objective To investigate the roles and mechanisms of action phosphorylated extracellular signal-regulated kinasel/2 （p-ERK1/ 2） and phosphorylated cAMP response element binding protein （p-CREB） in exogenous sodium hydrosulfide （NailS）-induced neuroprotec- tion after cerebral ischemia-reperfusion （I/R） in rats. Methods A total of 180 male Spragne-Dawley rats were randomly and equally divided into sham-operation group, model group, and NariS （ 100 μmol/L） intervention group. The rat model of middle cerebral artery occlusion was established by intraluminal suture technique. Reperfusion was performed after 2 hrs. Rats were given saline solution or NariS by intraperito- neal injection 20 min before reperfusion. At 6 hrs, 1 day, 2 days, 5 days, and 7 days after surgery, triphenyltetrazolium chloride staining was used to observe volume of cerebral infarction; Nissl staining was used to evaluate apeptosis in neurons. Western blot was used to determine the expression of p-ERK1/2 and p-CREB in the hippocampus; immunohistochemical analysis was used to measure the expression of B cell lymphoma/lewkmia-2 （Bcl-2）. Results Compared with the model group, the volume of cerebral infarction and apoptosis in neurons were significantly reduced in the NariS intervention group （P 〈 0.05）. At each time point, the protein expression of p-ERK1/2 and p-CREB was significantly higher in the NariS intervention goup than in the model group （P 〈0. 05 ）. The expression of p-ERK1/2 was positively correla- ted with the expression of p-CREB. Moreover, the NariS intervention group had significantly higher expression of Bcl-2 than the model group （P 〈0. 05 ）. Conehtsions After cerebral I/R injury in rats, Naris may protect neurons against apoptosis and reduce volmne of cerebral in- farction by inducing phosphorylation of ERK1/2 and CREB and stimulating expression of downstream Bcl-2.

关 键 词：脑 缺血再灌注损伤 硫化氢 细胞外调节蛋白激酶 CAMP反应元件结合蛋白 B细胞淋巴瘤/白血病-2 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]