精胺预处理对离体灌流大鼠心肌缺血/再灌注损伤及细胞凋亡的影响  

Effect of spermine preconditioning on myocardial ischemia/reperfusion injury and cardiomyocyte apoptosis in isolated perfused rat heart

作　　者：赵雅君[1] 王艳丽[1] 杜丽娟[1] 孙智睿[1] 张伟华[1] 王丽娜[1] 薛过[1] 王伟伟[1] 张昊[1] 张力[1] 

机构地区：[1]哈尔滨医科大学病理生理学教研室,黑龙江哈尔滨150081

出　　处：《中国药理学通报》2012年第8期1135-1140,共6页Chinese Pharmacological Bulletin

基　　金：黑龙江省教育厅课题基金资助(No 11531076)

摘　　要：目的探讨精胺预处理对离体灌流大鼠心肌缺血/再灌注损伤的影响及其可能的抗凋亡作用。方法应用Langehdorff离体灌流大鼠心脏复制模拟心肌缺血/再灌注损伤模型。24只大鼠随机分为3组,每组8只:对照组(Con-trol)、缺血/再灌注组(IR)、精胺干预组(Spermine)。比色法测定冠脉流出液中乳酸脱氢酶(LDH)活力,心肌组织中超氧化物歧化酶(SOD)活性及心肌丙二醛(MDA)含量;多导生理记录系统记录心脏功能;HE染色光镜下观察心肌形态学变化;三苯基氯化四氮唑(TTC)染色检测心肌梗死面积;透射电镜和TUNEL方法检测细胞凋亡;免疫组化检测心肌Fas与Bcl-2蛋白的表达。结果 (1)与对照组比,IR组冠脉LDH漏出明显增多、心肌组织中MDA水平增加、SOD活性降低(P<0.01);心功能明显下降(LVDP,HR,CF均低于对照组,P<0.05);光镜下可见心肌细胞呈凝固性或带状坏死。与Control组比,IR组心肌梗死面积及心肌细胞凋亡率明显增加(P<0.01);心肌Fas蛋白表达上调,Bcl-2蛋白表达下调。透射电镜下心肌细胞核染色质浓缩、凝聚且边集,染色质在核膜周边聚集形成新月形,线粒体嵴排列紊乱。(2)与IR组比,Spermine组冠脉LDH漏出减少,心肌组织中MDA减少,SOD增加(P<0.01);心功能有明显改善(LVDP,HR,CF均明显高于IR组,P<0.05);光镜下心肌细胞结构清晰。Spermine组与IR组比心肌梗死面积及心肌细胞凋亡率均明显降低(P<0.01);心肌Fas蛋白表达下调,Bcl-2蛋白表达上调;电镜下可见心肌肌节结构清晰,线粒体嵴完整、基质致密,未见核染色质凝聚。结论外源性精胺能明显减轻离体灌流大鼠心肌缺血/再灌注损伤,其机制可能与精胺抑制细胞凋亡有关。Aim To explore the effect of spermine preconditioning on myocardial ischemia/reperfusion injury and its potential antiapoptotic action in isolated perfused rat heart. Methods The model of myocardial is- chemia-reperfusion injury was replicated with Langehdorff isolated perfused rat heart. 24 rats were randomly divided into 3 groups： control, IR and spermine, with 8 rats in each group. LDH activity in coronary effluent, SOD activity and MDA content in myocardium were determined with colorimetry. Cardiac perform- ances were monitored on multichannel physiologic recorder. Morphological changes of myocardium were ob- served by light microscope with HE staining. Myocardial infarction size was measured with TTC staining. Cell apoptosis was detected with transmission electron microscope and TUNEL assay. Fas and Bcl-2 protein expressions were tested with immunohistochemistry. Results （1） Compared with the control group, in IR group, LDH activity in coronary effluent increased dis- tinctly, MDA in myocardial tissues rose, SOD activity decreased （P 〈 0. 01 ）, and heart function declined obviously（ LVDP, HR, CF were all lower than those in the control group, P 〈 0.05 ）. Myocardial cells were found to show coagulated zonal necrosis under light microscope. In comparison to the control, in IR group, myocardial infarction size and myocardial ceils apoptotic rate increased evidently （ P 〈 0. 05 ）, Fas protein ex- pression was up-regulated while Bcl-2 expression down- regulated. Under transmission electron microscope, myoeardium ceils displayed chromatin condensation and aggregation at the periphery of the nucleons, with chromatins aggregating like membrane and mitochondria a crescent around nuclear crista disorder. （2） Cornpared with IR group, in spermine group, LDH activity decreased, MDA content declined, while SOD activity increased（ P 〈 0.01 ） , heart function improved greatly （LVDP, HR and CF were all higher than those in IR group, P 〈 0. 05 ）. Cardiac myoeytes showed clear structure

关 键 词：精胺 缺血/再灌注 心脏 坏死 凋亡 FAS Bcl-2 大鼠 

分 类 号：R332[医药卫生—人体生理学] R322.11[医药卫生—基础医学]