SP600125对脑死亡大鼠心肌细胞凋亡的保护作用  

Protective role of SP600125 in apoptosis of cardiomyocytes in brain death rats

作　　者：王伟涛[1] 曹胜利[1] 方红波[1] 郭文治[1] 李捷[1] 阎冰[1] 张水军[1] 

机构地区：[1]郑州大学第一附属医院肝胆胰外科,450052

出　　处：《中华实验外科杂志》2015年第4期805-807,共3页Chinese Journal of Experimental Surgery

基　　金：河南省科技厅科技攻关计划资助项目，郑州市创新团队项目

摘　　要：目的 观察c-Jun氨基末端激酶（JNK）信号传导通路特异阻断剂SP600125对脑死亡状态下大鼠心肌细胞凋亡的保护作用.方法 将20只健康雄性大鼠随机分为4组,每组5只.空白对照（Sham）组：仅给予颅内置管,不加压诱导脑死亡;脑死亡（BD）组：仅诱导大鼠脑死亡并维持脑死亡6 h;SP600125组：在诱导脑死亡前1h腹腔注射SP600125（10 mg/kg）,并维持脑死亡6h;二甲基亚砜（DMSO）组：在诱导脑死亡前1h腹腔注射DMSO（10 mg/kg）,并维持脑死亡6h.应用实时定量聚合酶链反应（Real-time PCR）和Western blot检测各组心肌细胞中半胱氨酸天冬氨酸特异性蛋白酶-3（Caspase-3）与细胞色素C（Cyt-C）mRNA和蛋白的表达,并用原位缺口末端标记法（TUNEL）染色比较各组心肌细胞凋亡.结果 （1）Real-time PCR结果显示BD组Caspase-3mRNA和Cyt-C mRNA表达（2.37 ±0.12、2.03±0.08）显著高于Sham组（P ＜0.05）;BD组Caspase-3 mRNA和Cyt-C mRNA表达与DMSO组（2.38±0.11、2.06±0.09）接近（P＞0.05）;SP600125组Caspase-3mRNA和Cyt-C mRNA表达（1.21±0.05、1.23±0.24）显著低于BD组（P＜0.05）.（2）Western blot结果显示,BD组Caspae-3和Cyt-C表达（145.63±7.21、73.67±7.35）比Sham组（38.86±8.95、18.74±8.77）显著增多（P＜0.05）;BD组Caspase-3和Cyt-C表达与DMSO组（146.56±6.87、72.45±6.84）接近（P ＞0.05）;SP600125组Caspase-3和Cyt-C表达（39.45±5.73、20.34±5.69）显著低于BD组（P＜0.05）.（3） TUNEL法结果显示,BD组心肌细胞凋亡率[（26.39±4.99）％]比Sham组[（1.41±0.35）％]显著增加（P＜0.05）,DMSO组[（26.28±4.92）％]与Sham组凋亡率接近（P＞0.05）,SP600125组[（7.63±3.09）％]比BD组显著降低（P＜0.05）.结论 SP600125通过特异性地抑制JNK活性显著地减轻脑死亡状态下大鼠心肌细胞凋亡.Objective To investigate the protective role of SP600125,a selective c-Jun N-terminal kinase inhibitor,in the apoptosis of cardiomyocytes in brain death rats.Methods Twenty Sprague-Dawley （SD） rats were randomly divided into：Sham group （n =5）,only given intracranial insertion of Fogarty tube after anesthesia; brain-death group （n =5）,given brain death for 6 h ; SP600125 group （n =5）,given peritoneal injection of SP6600125 （10 mg/kg） 1 h before brain death for 6 h; and DMSO group （n =5）,given peritoneal injection of placebo solution DMSO （10 mg/kg） 1 h before brain death for 6 h.At 6 h,the apical myocardial samples were obtained for detection of Cysteinyl aspartate-specific protease （Caspase）-3 and cytochrome-C （Cyt-C） mRNA and protein expression by real-time quantitative polymerase chain reaction （real-time PCR） and Western blotting respectively.Apoptosis of cardiac muscle cells was determined by terminal deoxynucleotidyl transferase dUTP nick end labelling （TUNEL） assay.Results （1） Real-time PCR showed that the mRNA expression of Caspase-3 and Cyt-C mRNA in brain death group was （2.37 ±0.12,2.03 ±0.08） times higher than that in sham group （P 〈0.05）.There was no significant difference in the mRNA expression of Caspase-3 and Cyt-C between brain death group and DMSO group （2.38 ± 0.11 and 2.06 ± 0.09） （P 〉 0.05）.The mRNA expression of Caspase-3 and Cyt-C in SP600125 group （1.21 ±0.05 and 1.23 ±0.24） was significantly lower than that in brain death group （2.38 ± 0.11 and 2.06 ± 0.09） （P 〈 0.05）.（2） Western blotting revealed that the expression of Caspase-3 and Cyt-C in brain death group （145.63 ± 7.21 and 73.67 ± 7.35） was increased significantly as compared with that in Sham group （38.86 ± 8.95 and 18.74 ± 8.77,P 〈 0.05）.The expression of Caspase-3 and Cyt-C in brain death group was similar to that in DMSO group （146.56 t 6.87 and 72.45 ±6.84,P〉0.05）,and that in SP600125 group （39.45 ±5.73 and 20.

关 键 词：供体心脏 脑死亡 脱噬作用 c-Jun氨基末端激酶抑制剂 

分 类 号：R965[医药卫生—药理学]