心脏撞击伤后心肌细胞钙超载与结蛋白破坏的试验-第三军医大学学报.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 黄一 闵家新 熊健（400037重庆 第三军医大学新桥医院胸外科） [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[(110.4±17.6) vs (74.0±9.3), P0.01] ，伤后4h达最高值(141.1±17.8)后逐渐下降，48h恢复至伤前水平(78.6±10.3)；② MC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[(0.11±0.11) vs (1.23±0.27), 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[(110.4±17.6) vs (84.6±9.7), 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8h，D组高于T组[(0.55±0.18) vs (0.11±0.11),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，随着胞内Ca2+浓度增高，其介导的心肌结蛋白降解增加，通过伤后应用钙通道阻滞剂降低Ca2+浓度可有效减缓解结蛋白的降解破坏。 [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 , XIONG Jian (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[(110.4±17.6) vs (74.0±9.3),P0.01], and gradually decreased the highest in 4h after injury(141.1±17.8), there are normal in 48h after injury(78.6±10.3). ②The protein expression of desmin was lower than control group afte