B细胞淋巴瘤／白血病-2过表达对大鼠全脑缺血／再灌注后海马回磷酸化细胞外信号调节激酶的影响  

Effect of overexpression of B-cell lymphoma/leukemia-2 on the expression of phosphor-extracellular signal-regulated kinase in hippocampus of rats after global cerebral ischemia/reperfusion

作　　者：雷晓鸣[1] 吴刚[1] 吕建瑞[1] 薛荣亮[1] 

机构地区：[1]西安交通大学医学院第二附属医院麻醉科,710004

出　　处：《国际麻醉学与复苏杂志》2012年第11期739-743,共5页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金（30070731）;陕西省科技攻关课题：2010k15-03（1）,（2009K16-02）

摘　　要：目的观察B细胞淋巴瘤／白血病-2（B-cell lymphoma／leukemia-2，Bcl-2）过表达对大鼠全脑缺血／再灌注（ischemia/reperfusion，I／R）后磷酸化细胞外信号调节激酶（phosphor-extracellular signal-regulated kinase，p-ERK）蛋白在海马回表达的影响。方法90只健康雄性SD大鼠采用随机数字表法分为：假手术（SO）组，I／R组，Bcl-2过表达（Bcl-2）组。采用改良四血管法（four vessels occlusion method，4-VO）建立全脑堋模型。应用HE染色，免疫组化染色及原位末端标记（terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling，TUNEL）方法观察海马回神经元形态改变，凋亡细胞及p-ERK在CA1区和CA3区的不同表达。结果HE染色显示I／R组再灌注后48h CA1区神经元数目减少，排列紊乱，核膜界限不清，结构模糊，CA3区较CA1区变化轻微；Bcl-2组变化不明显。TUNEL染色显示，SO组可见到少量凋亡细胞；I／R组再灌注后48h凋亡细胞数达高峰，CA1区（110±13）明显多于CA3区（145±18）（P〈0．05）；Bcl-2组较I／R组凋亡细胞数量明显减少（CA1区：143±15，CA3，区：165±10）（P〈0．05）。免疫组化染色显示：p-ERK在SO组CA1区、CA3区的表达基本呈阴性；豫组再灌注后2h于CA3区开始弱表达，24h达高峰，然后逐渐下降，CA1区（150±14）表达弱于CA3区（125±9）（P〈0．05）；Bcl-2组表达明显强于I／R组（CA1区：123±13，C山区：100±10）（P〈0．05）。结论Bcl-2过表达可增加脑非后海马区p-ERK的表达，抑制细胞凋亡，其抗凋亡机制与ERK信号转导通路有关。Objective To explore the effect of B-cell lymphoma/leukemia-2（Bcl-2） overexpression on phosphor-extracellular signal-regulated kinase （p-ERK） expression in the hippocampus of rats after global cerebral ischemiMrepeffusion （I/R）. Methods 90 healthy male SD rats were randomly divided into sham opration group （SO group, n=30）, I/R group （n=30） and Bcl-2 overexpression group （Bcl-2 group, n=30）. Global cerebral I/R model was induced by four vessels occlusion method （4-VO）. The neuronal morphology changes, apoptosis and p-ERK expression in CA1 and CA3 was determined by HE staining, terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling （TUNEL） staining and immunohistochemical staining, respectively. Results HE staining showed the number of neuron in CA1 of I/R group decreased at 48 h after global cerebral I/R, combined with neuron disarrangement, nucleus membrane indistinction and nucleolus disappearance. The above change in CA3 was slighter than that in CA1 and the those changes in Bcl-2 group was not obvious. TUNEL staining showed there was few apoptosis in SO group. The number of apoptotic cells peaked 48 h after global cerebral I/R and the number of apoptotic cells in CA1 （110±13 ） was much more than that in CAn （145±18）（P〈0.05）. While the apoptotic cells in Bcl-2 group was much less than that in I/R group （CA1： 143±15, CA3： 165±10）（P〈0.05）. Immunohistochemical staining showed that p-ERK expression in CA1 and CA3 was almost negative in SO group. In I/R group p-ERK expression weaked in CA3 2 h after global cerebral I/R, peaked 24 h, then decreased; and p-ERK expression in CA1（150±14） was weaker than that in CA3（125±9）（P〈0.05）. While p-ERK expression in Bcl-2 group（CA1： 123±13, CA3： 100±10） was much stronger than that in I/R group （P〈0.05）. Conclusions Overexpression of Bcl-2 increased p-ERK expression while dramatically alleviated apoptosis. The mechanism of its antiapoptotic effect is related to ERK sign

关 键 词：缺血 再灌注 B细胞淋巴瘤 白血病-2 转基因 磷酸化细胞外信号调节激酶 

分 类 号：R733[医药卫生—肿瘤]