调控沉默信息调节因子2同系物1活性对脑缺血大鼠血管发生的影响  

Effect of silent information regulator 2 homologue 1 activity on angiogenesis in cerebral ischemia rats

作　　者：马毓[1] 何晓燕[1] 党辉[1] 朱沂[1] 李红燕[1] Ma Yu;He Xiaoyan;Dang Hui;Zhu Yi;Li Hongyan(Department of Neurology, People＇ s Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830001, China)

机构地区：[1]新疆维吾尔自治区人民医院神经内科,乌鲁木齐830001

出　　处：《中华神经科杂志》2018年第6期456-463,共8页Chinese Journal of Neurology

基　　金：新疆维吾尔自治区自然科学基金资助项目（2014211A057）

摘　　要：目的探讨调控沉默信息调节因子2同系物1（SIRT1）活性对脑缺血大鼠血管发生的影响及其相关机制.方法将120只清洁级健康雄性SD大鼠,采用随机数字表法随机分为4组：假手术组、脑缺血再灌注损伤模型组（模型组）、SIRT1激动剂组（激动剂组）和SIRT1抑制剂组（抑制剂组）,每组大鼠各30只.用线栓法制作短暂性大脑中动脉栓塞模型;恢复血流再灌注24h,对大鼠进行神经功能评分;氯化三苯基四氮唑染色法测定大鼠脑梗死面积;ELISA法检测缺血脑组织SIRT1去乙酰化酶活性;免疫组织化学法检测缺血脑皮质区CD34表达和微血管密度（MVD）计数;蛋白质印迹法检测缺血脑组织SIRT1、血管内皮生长因子（VEGF）和促红细胞生成素（EPO）蛋白伪表达水平.结果模型组缺血脑组织SIRT1去乙酰化酶活性为（7.721±0.581）U/L,较假手术组[（13.828±0.828）U/L]显著降低（t=8.650,P〈0.01）;激动剂组缺血脑SIRT1去乙酰化酶活性为（26.165±0.971）U/L,较模型组显著升高（t=-26.123,P〈0.01）;抑制剂组缺血脑SIRT1去乙酰化酶活性（17.094±1.012）U/L,较激动剂组显著降低（t=12.848,P〈0.01）.大鼠缺血脑组织SIRT1被激活后,其神经功能缺损评分[（1.333±0.516）分]较模型组[（2.667±0.516）分]明显降低（t=4.822,P〈0.01）;抑制SIRT1活性后,其神经功能缺损评分[（2.500±0.548）分]较激动剂组显著升高（t=-4.147,P〈0.01）.模型组脑梗死面积为15.473％±3.049％,激活缺血脑组织SIRT1后,脑梗死面积（9.152％±1.803％）明显缩小（t=3.188,P〈0.05）.免疫组织化学结果显示缺血脑皮质区染成棕色的细胞为CD34染色阳性细胞;激活SIRT1可使缺血脑皮质区MVD计数显著增加[激动剂组MVD计数：每高倍镜下（8.167±1.941）个,模型组MVD计数：每高倍镜下（3.167±0.753）个,t=-6.864,P〈0.01].免疫印迹结果表明,激活缺血脑组织SIRT1后,缺血脑组织中VEGF（激动剂组：Objective To investigate the effect of silent information regulator 2 homologue 1 （SIRT1） activity on angiogenesis in cerebral ischemia rats and its related mechanisms .Methods One hundred and twenty healthy male Sprague-Dawley rats were randomly divided into four groups by random numbers：control group , the cerebral ischemia-reperfusion injury model group （ model group ） , the SIRT1 agonist group （agonist group） and the SIRT1 inhibitor group （inhibitor group）, with 30 rats in each group. A model of transient middle cerebral artery occlusion was performed by the suture method .After reperfusion for 24 h, neurological deficit scores were evaluated . Cerebral infarct area after middle cerebral artery occlusion （ MCAO） in rats was determined by staining with triphenyltetrazolium chloride .SIRT1 deacetylase activity was detected by ELISA in ischemic brain tissue.By immunohistochemistry, we observed CD34 expression and detected microvascular density （ MVD） in ischemic cerebral cortex.Immunoblotting was carried out to evaluate protein levels of SIRT 1, vascular endothelial growth factor （VEGF） and erythropoietin （EPO） in ischemic brain tissue.Results Compared with the control group （（13.828 ±0.828） U/L）, SIRT1 deacetylase activity was significantly reduced in ischemic brain in the model group （（7.721 ±0.581） U/L,t=8.650,P〈0.01）.Compared with the model group , SIRT1 deacetylase activity was significantly increased in ischemic brain in the agonist group （（26.165 ±0.971） U/L,t=-26.123,P〈0.01）. Compared with the agonist group , SIRT1 deacetylase activity was significantly reduced in ischemic brain in the inhibitor group （（17.094 ±1.012）U/L,t=12.848,P〈0.01）.Neurological deficit score was 2.667 ± 0.516 in the model group.When SIRT1 was activated in ischemic brain , neurological deficit score was significantly lower （1.333 ±0.516,t=4.822,P〈0.01） than that of the model group .When SIRT1 activity was inhibited, the neurological deficit score inc

关 键 词：脑缺血 再灌注损伤 沉默信息调节蛋白质类 神经保护 血管生成 血管内皮生长因子 红细胞生成素 

分 类 号：R743[医药卫生—神经病学与精神病学]