膜攻击复合物C5b-9对创伤失血性休克大鼠肝损害的影响研究  

The effect of the membrane attack complex C5b-9 on liver cells during traumatic hemorrhagic shock in rat

作　　者：赵志伶[1] 曹书华[1] 王勇强[1] 

机构地区：[1]天津医科大学第一中心临床学院,天津市第一中心医院急救医学研究所,300192

出　　处：《中国危重病急救医学》2011年第3期158-161,193,共5页Chinese Critical Care Medicine

基　　金：天津市医药卫生科研基金资助项目（05KYZ17）

摘　　要：目的 观察大鼠创伤失血性休克时肝脏是否受膜攻击复合物攻击,以及膜攻击复合物是否对肝脏细胞凋亡产生影响.方法 雄性Wistar大鼠50只,按随机数字表法均分为正常组及模型1、3、6、24 h组5组.采用骨折后经颈动脉放血至血压40 mm Hg（1 mm Hg=0.133 kPa）制备创伤失血性休克模型.取血浆,采用酶联免疫吸附法（ELISA）检测膜攻击复合物C5b-9浓度,采用速率法测定丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）浓度;取肝脏组织,采用免疫组化法检测C5b-9阳性表达,用原位末端缺刻标记法（TUNEL）检测肝细胞凋亡,苏木素-伊红（HE）染色,光镜下观察病理改变.结果 正常组血中能检测到少量的C5b-9;模型1、3、6 h组血中C5b-9浓度（ng/L）较正常组显著升高（272.91±9.56、192.01±9.04、156.78±8.37比25.98±5.87,均P＜0.05）.模型3 h组血ALT（U/L）、模型1 h组血AST（U/L）即显著上升（92.90±8.83、264.83±31.14）,24 h达高峰（184.30±12.98、647.36±60.02）,与正常组（38.75±5.40、66.69±19.95）比较差异均有统计学意义（均P＜0.05）.正常组及模型1 h、6 h组肝脏未发现C5b-9阳性表达;模型3 h组门管区大量肝脏实质细胞存在C5b-9（个）沉积（22.60±1.06）,模型24 h组C5b-9沉积明显减少（2.20±0.60,P＜0.05）.正常组未检测到凋亡细胞;模型1、6、24 h组发现散在凋亡细胞（个：1.20±0.25、5.60±0.37、1.60±0.26）,模型3 h组中央静脉周围凋亡细胞明显增加,出现凋亡高峰（20.60±0.47）,与其余各组比较差异均有统计学意义（均P＜0.05）.模型组可见肝细胞水肿变性、肝细胞膜完整性破坏,细胞溶解,以24 h病理损害最重.结论 创伤失血性休克时大鼠肝脏受到膜攻击复合物C5b-9的攻击,并且肝脏C5b-9的表达高峰与凋亡高峰在同一时间点出现,但并不是同一部位;模型3 h后血中低水平的C5b-9提示预后极差.Objective To observe whether the membrane attack complex C5b-9 would accumulate in the rats＇ liver after receiving the assault of traumatic hemorrhagic shock, and whether the membrane attack complex deals an impact on liver apoptosis. Methods Fifty male healthy Wistar rats were randomly divided into five groups： normal group, 1, 3, 6, 24-hour model groups. The model of traumatic hemorrhagic shock was reproduced by withdrawal of blood from carotid artery after a bone fracture till the blood pressure lowered to 40 mm Hg（1 mm Hg= 0. 133 kPa）. Plasma membrane attack complex C5b-9 concentration was assayed using enzyme linked immunoadsorbent assay. Alanine aminotransferase（ALT）and aspartate aminotransferase（AST）in blood was determined by Rate method. Immunohistochemistry was used to detect C5b-9 deposition in the liver. Apoptosis of liver cells was then detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling（TUNEL）assay. The pathological changes in paraffin sections stained with hematoxylin-eosin（HE）were observed under light microscope. Results A small amount of C5b-9 in plasma was found in normal group, and the values（ng/L）of 1, 3, 6-hour models were significantly higher than those of the normal group（272. 91±9.56, 192. 01±9. 04, 156. 78±8. 37 vs. 25. 98±5.87, all P〈0. 05）. ALT（U/L）in 3-hour model group and AST（U/L）in 1-hour model group were increased significantly（92.90 ± 8. 83, 264.83 ± 31.4）, peaked at 24 hours（184.30 ± 12. 98, 647.36 ± 60. 02）, and there was significant difference compared with normal group（38. 75±5.40, 66. 69± 19.95, all P〈0. 05）. In the normal group and the 1-hour and 6-hour model groups, no C5b-9 was found in liver, but in the 3-hour model group a large number of liver parenchymal cells in the portal area were found to contain C5b-9 （22. 60± 1.06）, however the number decreased significantly in the 24-hour model（2. 20±0. 60, P〈0. 05）.In normal group there was no apoptotic cell, and in 1,

关 键 词：休克 创伤失血性 肝脏 膜攻击复合物 凋亡 

分 类 号：R459.7[医药卫生—急诊医学]