心脏撞击伤后心肌细胞钙超载与结蛋白破坏的实验研究.DOC

大鼠心肌挫伤后细胞结蛋白破坏与钙超载的实验研究 陈小波 余祖滨 闵家新 黄一 熊健（第三军医大学新桥医院胸外科 重庆 400037） [摘要] 目的 研究大鼠心肌挫伤（Myocardial Contusion MC）后细胞结蛋白的破坏与钙超载情况，评价心肌挫伤后结蛋白（desmin）破坏与钙超载的关系。方法：SD大鼠随机分为标准对照组（S组）、单纯致伤组（T组）和致伤+地尔硫卓组（D组），BIM-Ⅲ生物撞击机建立胸部撞击致心肌挫伤模型，致伤后D组腹腔注射盐酸地尔硫卓(0.5mg/kg)，分别于伤前、伤后2h、4h、8h、24h、48h检测心肌细胞钙离子浓度，免疫组化法及western-blot检测心肌细胞结蛋白变化情况，RT-PCR检测结蛋白mRNA表达变化情况。结果 ①伤后2h心肌细胞Ca2+浓度明显增高（P0.01），伤后4h达最高值后逐渐下降，48h恢复至伤前水平；②MC后早期即出现严重的结蛋白破坏，伤后8h结蛋白表达量降至最低（P0.01），24h开始部分恢复，伤后48h结蛋白表达增高且出现排列紊乱和无规则聚集的现象。③与T组比较，D组显著降低了伤后4、8h胞内Ca2+浓度（P0.01），结蛋白表达量在伤后2h、4h两组相较无明显差异，但在伤后8hD组高于T组（P0.05）④结蛋白mRNA伤后表达增高，24h达峰值，D组24h后表达低于T组。结论：MC后早期结蛋白的破坏以细胞的机械损伤为主，伤后胞内Ca2+浓度增高介导的结蛋白降解加重了细胞的损伤，伤后应用钙通道阻滞剂能够减缓钙超载对结蛋白的破坏作用。 [关键词] 创伤；心肌挫伤；结蛋白；钙超载；钙拮抗剂 [中图法分类号A study of desmin degradation and Ca2+ overload after the Myocardial contusion in rats CHEN Xiaobo，YU Zubin ，MIN Jiaxin，HUANG Yi (Department of Thoracic Surgury, Xinqiao Hospital ,Third Military University ,Chongqing　400037 ,China) [Abstract] Objective To study the relationship of desmin degradation and Ca2+ overload after the Myocardial Contusion (MC). Methods SD rats were randomly divided into trauma group (T), trauma+ diltiazem group (D), and control group (S). The models of MC were established with BIM-Ⅲ Bio-impactor, animal of D group were treated with intraperitoneal injection of diltiazem (0.5mg/kg) after injury. Western-blot analysis, immunohistochemisty and RT-PCR were used to detect the quantitative expression of desmin protein, free Ca2+ content detected by confocal laser scanning microscope in 2h、4h、8h、24h、48h after injury. Results ①The free Ca2+ in myocardial cells was higher in 2h after injury（P0.01）, and gradually decreased the highest in 4h after injury, there are normal in 48h after injury.②The protein expression of desmin was lower than control group after injury, diminishing degree of desmin was significant in 8h after injury, the protein were aggregated random in 48h after injury.③Decreased free Ca2+ content 4h、8h after injury compare with T group (P0.01), a