CAJ | 학술논문

目的探讨JNK特异性抑制剂-SP600125对大鼠脑缺血/再灌注(schemia/reperfusion,I/R)中海马细胞凋亡的作用.方法雄性SD大鼠54只,体重量230 g～250 g,采用双盲随机方法分成假手术组(SH组),I/R组,JNK抑制剂SP600125组(SP组),每组根据再灌注时间分为30 min、24 h和72h 3个亚组,每亚组6只动物.采用4-VO法建立SD大鼠脑缺血模型,于缺血前30 min侧脑室注射二甲基亚砜(dimethyl sulfoxide,DMSO),DMSO及JNK抑制剂SP600125(溶媒采用DMSO),容积均为10μl.脑I/R后24、72 h测定其行为学改变,分别在30 min、24 h和72 h等时间点,取脑组织,免疫组织化学方法测定海马CA1区Bcl-2和Bax蛋白表达阳性细胞数量和凋亡锥体细胞数量.结果全脑I/R使大鼠的垂直运动次数(4.8±2.0,9.1±3.4)和平衡木积分(2.3±1.2,3.5±0.9)显著减少(P＜0.05),I/R组的减少最为显著 ;脑I/R后海马CA1区凋亡神经元细胞数目(40.5±5.1)显著低于I/R组(P＜0.01) ;全脑I/R 24 h后,海马CA1区Bcl-2和Bax阳性锥体细胞数目(89.7±8.4,40.5±2.3)显著增加(P＜0.01),再灌注24 h组增加最多 ;SP600125可以增加Bcl-2蛋白表达、减少Bax蛋白表达.结论 SP600125对大鼠脑I/R中海马细胞的凋亡具有保护作用,此机制涉及抑制JNK信号转导通路.
목적 탐토JNK특이성억제제-SP600125대대서뇌결혈/재관주(schemia/reperfusion,I/R)중해마세포조망적작용.방법 웅성SD대서54지,체중량230 g～250 g,채용쌍맹수궤방법분성가수술조(SH조),I/R조,JNK억제제SP600125조(SP조),매조근거재관주시간분위30 min、24 h화72h 3개아조,매아조6지동물.채용4-VO법건립SD대서뇌결혈모형,우결혈전30 min측뇌실주사이갑기아풍(dimethyl sulfoxide,DMSO),DMSO급JNK억제제SP600125(용매채용DMSO),용적균위10μl.뇌I/R후24、72 h측정기행위학개변,분별재30 min、24 h화72 h등시간점,취뇌조직,면역조직화학방법측정해마CA1구Bcl-2화Bax단백표체양성세포수량화조망추체세포수량.결과 전뇌I/R사대서적수직운동차수(4.8±2.0,9.1±3.4)화평형목적분(2.3±1.2,3.5±0.9)현저감소(P＜0.05),I/R조적감소최위현저 ;뇌I/R후해마CA1구조망신경원세포수목(40.5±5.1)현저저우I/R조(P＜0.01) ;전뇌I/R 24 h후,해마CA1구Bcl-2화Bax양성추체세포수목(89.7±8.4,40.5±2.3)현저증가(P＜0.01),재관주24 h조증가최다 ;SP600125가이증가Bcl-2단백표체、감소Bax단백표체.결론 SP600125대대서뇌I/R중해마세포적조망구유보호작용,차궤제섭급억제JNK신호전도통로.
Objective To investigate the effect of SP600125 on cell apoptosis in hippocampal CA1 after cerebral ischemia/reperfusion (I/R) in rats and its possible mechanism.Methods 54 SD rats weighing 230 g-250 g were randomly divided into sham operation group (SH),I/R group and JNK inhibitor SP600125 group (SP).The rats received intracerebroventricularly DMSO(SH group),DMSO(I/R group) and SP600125(SP group,the use of DMSO solvent) 30 min before ischemia.Each group was divided into 3 subgroups according to reperfusion time 30 min,24 h and 72 h (6 animals for each subgroup).Global cerebral ischemia was induced by four-vessel occlusion in rats.Behavioral test was performed at 24 h and 72 h after cerebral ischemia/reperfusion.Bax and Bcl-2 positive pyramidal cells as well as apoptotic positive pyramidal cells in the CA1 were quantified in immunohistochemical stained or terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) stained sections.Results Compared with SH group,cerebral I/R induced a decrease in standing times(4.8±2.0 vs 9.1±3.4,P＜0.05) and the beam-walking test scores(2.3±l.2 vs 3.5±0.9,P＜0.05).Immunohistochemistry results showed that after ischemia/reperfusion Hippocampal CA1 area Bcl-2 and Bax positive expression increased the number of pyramidal cells,24 h of reperfusion the number of pyramidal cells shows that the expression of positive to the peak(40.5±5.1)(P＜0.01),After I/R 24 h to 72 h,Positive expression to reduce the number of pyramidal cells (P＜0.05),With the IR group,SP group Bcl-2-positive pyramidal cells increased the number of (89.7±8.4)(P＜0.05),Baxpositive increase in the number of pyramidal cells less (40.5±2.3)(P＜0.05).Conclusions SP600125 protects hippocampal CA1 neurons from death during global brain ischemia/reperfusion injury through inhibition of JNK signal transduction pathway.