[Gly14]-Humanin对大鼠局灶性脑缺血再灌注损伤后炎症反应及细胞凋亡的影响  

Effect of [Gly14]-Humanin on inflammation and cell apoptotic in brain tissue after focal cerebral ischemia-reperfusion in rats

作　　者：余智 顾苏兵[2] 于民[2] 邵晓莉 洪小琴 姜慧华 吴燕飞 

机构地区：[1]淳安县第一人民医院康复医学科,311700 [2]浙江省人民医院神经内科 [3]淳安县第一人民医院神经内科,311700

出　　处：《浙江医学》2017年第22期1976-1980,共5页Zhejiang Medical Journal

基　　金：浙江省医药卫生科技项目(201482575);杭州市科技发展计划项目(20140633B66)

摘　　要：目的探讨Humanin衍生物([Gly14]-Humanin,HNG)预处理对大鼠局灶性脑缺血再灌注损伤后炎症反应、细胞凋亡的影响及其机制。方法将96只健康雄性SD大鼠按随机数字法分为HNG组、生理盐水组、模型组及假手术组,每组24只。采用改良的Zea-longa线栓法制备大脑中动脉缺血再灌注损伤模型,HNG组大鼠术前5 d给予HNG(2μl/kg),生理盐水组与假手术组大鼠予0.9%氯化钠溶液(2 ml/kg),均为股静脉注射,1次/d;模型组不作处理。各组大鼠在再灌注24 h后进行神经功能缺损评分(NDS),采用ELISA法测定大鼠脑组织核因子-κB p65(NF-κB p65)及干扰素-γ(IFN-γ)水平,原位末端标记染色观察凋亡细胞数及尼氏染色检测健存神经细胞数。结果 HNG组、生理盐水组、模型组及假手术组NDS分别为1.5(1.0,2.0)分、3.0(2.0,3.0)分、3.0(2.0,3.0)分和0.0(0.0,0.0)分,假手术组NDS低于另外3组,HNG组NDS低于模型组和生理盐水组(均P<0.05)。与假手术组比较,HNG组、生理盐水组及模型组大鼠的NF-κB p65水平[(13.48±0.85)、(22.15±0.96)、(22.62±0.64)pg/mg],IFN-γ水平[(5.17±0.67)、(10.83±0.34)、(10.81±0.19)p g/mg]及凋亡细胞的百分率[(48.09±7.62)%、(70.19±8.33)%、(69.02±9.07)%]均较高,不重叠高倍镜视野中尼氏小体的数量[(10.57±0.51)、(5.16±0.56)和(5.65±0.98)个]较少,差异有统计学意义(P<0.05);HNG组大鼠的NDS、NF-κB p 65和IFN-γ水平及细胞凋亡率均低于生理盐水组及模型组,尼氏小体数多于生理盐水组及模型组,差异有统计学意义(P<0.05);HNG组大鼠IFN-γ与NF-κB p65水平呈正相关(r=0.873,P<0.01)。结论 [Gly14]-Humanin可抑制脑缺血再灌注损伤过程中的局部炎症反应、减少细胞的凋亡,从而减轻临床神经功能的缺损。其机制可能与下调脑组织NF-κB p65的释放,进一步减少IFN-γ的表达有关。Objective To investigate the effect of [Gly14]-Humanin（HNG）on inflammation and cell apoptosis in brain tissue after focal cerebral ischemia-reperfusion injury in rats. Methods Ninety-six male Sprague-Dawley rats were randomly divided into sham-operation group, model group, normal saline（NS） group and HNG group with 24 rats in each group. The ischemia-reperfusion model was induced by middle cerebral artery occlusion（MCAO）with Zea-longa suture embolism method in rats. HNG（2μl/kg） or normal saline（2 ml/kg） were given to rats in HNG and NS group 5 days before operation（i.v. qd x5）, respectively. After 4.5 h of cerebral ischemia and 24 h of reperfusion, neurological deficit score（NDS）was assessed, the NF-κB p65 and IFN-γ levels in cerebral tissue were measured by ELISA,neuron apoptosis was detected by Nissl and TUNEL staining; the correlation between IFN-γ and NF-κB p65 levels in cerebral tissue was analyzed. Results The median NDS values in HNG, NS, model and sham-operation groups were 1.5（1.0, 2.0）, 3.0（2.0, 3.0）, 3.0（2.0, 3.0）and 0.0（0.0, 0.0）, respectively; the NDS of sham-operation group was significantly lower than those of other three groups, the NDS of HNG group was significantly lower than that of model group and normal saline group（all P0.05）. Compared with sham-operation group, the levels of NF-κB p65 in HNG group, normal saline group and model group（13.48±0.85, 22.15±0.96 and 22.62±0.64 pg/mg）, the levels of IFN-γ（5.17±0.67, 10.83±0.34 and10.81±0.19 pg/mg）, the ratio of apoptotic neurocyte（48.09±7.62, 70.19±8.33 and 69.02±9.07）were significantly increased, and the number of neurons（10.57±0.51, 5.16±0.56 and 5.65±0.98）was significantly decreased（all P0.05）. The NDS, NF-κB p65 and IFN-γ levels, ratio of apoptotic neurocytes in HNG group were significantly lower than those in NS group and model group; while the number of neurons was significantly higher than that in normal saline group and model group（all P0.05）.The l

关 键 词：[Gly14]-Humanin 脑缺血再灌注 核因子-ΚB P65 干扰素-Γ 细胞凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学]