中华麻醉学杂志
中華痳醉學雜誌
중화마취학잡지
CHINESE JOURNAL OF ANESTHESIOLOGY
2008年
5期
459-462
,共4页
细胞外信号调节MAP激酶类%细胞凋亡%再灌注损伤
細胞外信號調節MAP激酶類%細胞凋亡%再灌註損傷
세포외신호조절MAP격매류%세포조망%재관주손상
Extracellular signal-regulated MAP kinases%Apoptosis%Reperfusion injury
目的 探讨细胞外信号调节激酶(ERK)在脑缺血再灌注大鼠海马细胞凋亡中的作用.方法 健康雄性SD大鼠90只,体重280～320 g,随机分为3组(n=30):假手术组(S组)、脑缺血再灌注组(IR组)和ERK磷酸化特异性抑制剂PD98059组(PD组).采用4血管法建立大鼠脑缺血再灌注模型,于再灌注后2、6、12、24、48、72 h时,各取5只大鼠,断头取脑,光镜下观察海马CA1区和CA3区病理学结果 ,计算细胞凋亡指数(AI),采用免疫组化法检测磷酸化ERK(p-ERK)和磷酸化Bad(p-Bad)的表达.结果 与S组比较,IR组和PD组再灌注期间CAI区和CA3区AI升高,再灌注2、6、12 h时CA1区p-ERK表达降低,再灌注后CA1区和CA3区p-Bad表达降低(P＜0.05);与IR组比较,PD组再灌注后CA1医和CA3区AI升高,再灌注2、6、12、24 h时CA3区p-ERK表达降低,再灌注2、6 h时CA1区p-Bad表达降低,再灌注2、6、12h时CA3区p-Bad表达降低(P＜0.05).结论 脑缺血再灌注可降低ERK活性,导致Bad蛋白去磷酸化,从而诱发大鼠海马细胞凋亡.
目的 探討細胞外信號調節激酶(ERK)在腦缺血再灌註大鼠海馬細胞凋亡中的作用.方法 健康雄性SD大鼠90隻,體重280～320 g,隨機分為3組(n=30):假手術組(S組)、腦缺血再灌註組(IR組)和ERK燐痠化特異性抑製劑PD98059組(PD組).採用4血管法建立大鼠腦缺血再灌註模型,于再灌註後2、6、12、24、48、72 h時,各取5隻大鼠,斷頭取腦,光鏡下觀察海馬CA1區和CA3區病理學結果 ,計算細胞凋亡指數(AI),採用免疫組化法檢測燐痠化ERK(p-ERK)和燐痠化Bad(p-Bad)的錶達.結果 與S組比較,IR組和PD組再灌註期間CAI區和CA3區AI升高,再灌註2、6、12 h時CA1區p-ERK錶達降低,再灌註後CA1區和CA3區p-Bad錶達降低(P＜0.05);與IR組比較,PD組再灌註後CA1醫和CA3區AI升高,再灌註2、6、12、24 h時CA3區p-ERK錶達降低,再灌註2、6 h時CA1區p-Bad錶達降低,再灌註2、6、12h時CA3區p-Bad錶達降低(P＜0.05).結論 腦缺血再灌註可降低ERK活性,導緻Bad蛋白去燐痠化,從而誘髮大鼠海馬細胞凋亡.
목적 탐토세포외신호조절격매(ERK)재뇌결혈재관주대서해마세포조망중적작용.방법 건강웅성SD대서90지,체중280～320 g,수궤분위3조(n=30):가수술조(S조)、뇌결혈재관주조(IR조)화ERK린산화특이성억제제PD98059조(PD조).채용4혈관법건립대서뇌결혈재관주모형,우재관주후2、6、12、24、48、72 h시,각취5지대서,단두취뇌,광경하관찰해마CA1구화CA3구병이학결과 ,계산세포조망지수(AI),채용면역조화법검측린산화ERK(p-ERK)화린산화Bad(p-Bad)적표체.결과 여S조비교,IR조화PD조재관주기간CAI구화CA3구AI승고,재관주2、6、12 h시CA1구p-ERK표체강저,재관주후CA1구화CA3구p-Bad표체강저(P＜0.05);여IR조비교,PD조재관주후CA1의화CA3구AI승고,재관주2、6、12、24 h시CA3구p-ERK표체강저,재관주2、6 h시CA1구p-Bad표체강저,재관주2、6、12h시CA3구p-Bad표체강저(P＜0.05).결론 뇌결혈재관주가강저ERK활성,도치Bad단백거린산화,종이유발대서해마세포조망.
Objective To investigate the role of extracellular signal-regulated kinase in neuronal apoptosis of hippoeampus induced by global cerebral ischemia-reperfusion in rats.Methods Ninety healthy male SD rats weighing 280-320 g were randomly divided into 3 groups(n=30 each):groupI sham operation(S);groupⅡI/ R and group Ⅲ PD98059+I/R(PD).The animals were anesthetized with intraperitoneal 1% pentobarbital 40 mgkg.Global cerebral I/R was produced by 4-vessel occlusion method.Bilateral vertebral arteries were electrically cauterized and bilateral common carotid arteries were clamped for 5 min.Clamping was then released for reperfusion in group Ⅱ and Ⅲ.In group Ⅲ PD98059(a specific ERK inhibitor)O.3 mg/kg was injected iv before carotid artery clamping.Five animals in each group were sacrificed at 2,6,12,24,48 and 72 h of reperfusion and their brains were removed and cut into sections which were stained with HE and examined under microscope(400 times magnified).Neuronal apoptosis in hippieampal Cal and CA3 regions were detected by TUNEL assay.Apoptotic index(AI) was calculated.Phosphorylated ERK(p-ERK)and phosphorylated Bad (p-Bed)expression was assessed by immuno-histochemistry.Results (1)Hippoeampal pyramidal cells were orderly distributed and morphologically intact in group S but unevenly distributed with widened intercellular space,shrinked cell body and condensed nuclei in I/R group.The I/R induced changes were worse in PD group.(2)AI in Cal and CA3 region was significantly higher in PD group than in I/R group.(3)The p-ERK expression in Cal region was signiticanfly decreased at 2,6,12 h of reperfusion in I/R(Ⅲ)and PD groups(Ⅲ)as compared with group S(I),while p-ERK expression in CA3 region was significantly lower at 2,6,12,24 h of reperfusion in PD group(Ⅲ)than in S(I)and I/R group(Ⅱ).(4)The p-Bed expression in Cal and CA3 regio was signi ficantly lower during reperfusion in PD group than in I/R group and was lowest in group S(I).Conclusion Cerebral isehemia reperfusion can decrease the p-ERK expression and then leads to the dephosphorylation of Bed(a member of Bel-2 protein family),which induced the apeptosis in hippoeampal neurons.
