中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2011年
3期
158-161,后插1
,共5页
休克,创伤失血性%肝脏%膜攻击复合物%凋亡
休剋,創傷失血性%肝髒%膜攻擊複閤物%凋亡
휴극,창상실혈성%간장%막공격복합물%조망
Traumatic hemorrhagic shock%Liver%Membrane attack complex%Apoptosis
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.
目的 觀察大鼠創傷失血性休剋時肝髒是否受膜攻擊複閤物攻擊,以及膜攻擊複閤物是否對肝髒細胞凋亡產生影響.方法 雄性Wistar大鼠50隻,按隨機數字錶法均分為正常組及模型1、3、6、24 h組5組.採用骨摺後經頸動脈放血至血壓40 mm Hg(1 mm Hg=0.133 kPa)製備創傷失血性休剋模型.取血漿,採用酶聯免疫吸附法(ELISA)檢測膜攻擊複閤物C5b-9濃度,採用速率法測定丙氨痠轉氨酶(ALT)、天鼕氨痠轉氨酶(AST)濃度;取肝髒組織,採用免疫組化法檢測C5b-9暘性錶達,用原位末耑缺刻標記法(TUNEL)檢測肝細胞凋亡,囌木素-伊紅(HE)染色,光鏡下觀察病理改變.結果 正常組血中能檢測到少量的C5b-9;模型1、3、6 h組血中C5b-9濃度(ng/L)較正常組顯著升高(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組血ALT(U/L)、模型1 h組血AST(U/L)即顯著上升(92.90±8.83、264.83±31.14),24 h達高峰(184.30±12.98、647.36±60.02),與正常組(38.75±5.40、66.69±19.95)比較差異均有統計學意義(均P＜0.05).正常組及模型1 h、6 h組肝髒未髮現C5b-9暘性錶達;模型3 h組門管區大量肝髒實質細胞存在C5b-9(箇)沉積(22.60±1.06),模型24 h組C5b-9沉積明顯減少(2.20±0.60,P＜0.05).正常組未檢測到凋亡細胞;模型1、6、24 h組髮現散在凋亡細胞(箇:1.20±0.25、5.60±0.37、1.60±0.26),模型3 h組中央靜脈週圍凋亡細胞明顯增加,齣現凋亡高峰(20.60±0.47),與其餘各組比較差異均有統計學意義(均P＜0.05).模型組可見肝細胞水腫變性、肝細胞膜完整性破壞,細胞溶解,以24 h病理損害最重.結論 創傷失血性休剋時大鼠肝髒受到膜攻擊複閤物C5b-9的攻擊,併且肝髒C5b-9的錶達高峰與凋亡高峰在同一時間點齣現,但併不是同一部位;模型3 h後血中低水平的C5b-9提示預後極差.
목적 관찰대서창상실혈성휴극시간장시부수막공격복합물공격,이급막공격복합물시부대간장세포조망산생영향.방법 웅성Wistar대서50지,안수궤수자표법균분위정상조급모형1、3、6、24 h조5조.채용골절후경경동맥방혈지혈압40 mm Hg(1 mm Hg=0.133 kPa)제비창상실혈성휴극모형.취혈장,채용매련면역흡부법(ELISA)검측막공격복합물C5b-9농도,채용속솔법측정병안산전안매(ALT)、천동안산전안매(AST)농도;취간장조직,채용면역조화법검측C5b-9양성표체,용원위말단결각표기법(TUNEL)검측간세포조망,소목소-이홍(HE)염색,광경하관찰병리개변.결과 정상조혈중능검측도소량적C5b-9;모형1、3、6 h조혈중C5b-9농도(ng/L)교정상조현저승고(272.91±9.56、192.01±9.04、156.78±8.37비25.98±5.87,균P＜0.05).모형3 h조혈ALT(U/L)、모형1 h조혈AST(U/L)즉현저상승(92.90±8.83、264.83±31.14),24 h체고봉(184.30±12.98、647.36±60.02),여정상조(38.75±5.40、66.69±19.95)비교차이균유통계학의의(균P＜0.05).정상조급모형1 h、6 h조간장미발현C5b-9양성표체;모형3 h조문관구대량간장실질세포존재C5b-9(개)침적(22.60±1.06),모형24 h조C5b-9침적명현감소(2.20±0.60,P＜0.05).정상조미검측도조망세포;모형1、6、24 h조발현산재조망세포(개:1.20±0.25、5.60±0.37、1.60±0.26),모형3 h조중앙정맥주위조망세포명현증가,출현조망고봉(20.60±0.47),여기여각조비교차이균유통계학의의(균P＜0.05).모형조가견간세포수종변성、간세포막완정성파배,세포용해,이24 h병리손해최중.결론 창상실혈성휴극시대서간장수도막공격복합물C5b-9적공격,병차간장C5b-9적표체고봉여조망고봉재동일시간점출현,단병불시동일부위;모형3 h후혈중저수평적C5b-9제시예후겁차.
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, 6, 24-hour model groups there were scattered apoptotic cells(1. 20±0. 25, 5. 60±0. 37, 1. 60±0. 26). In the 3-hour model group apoptosis of hepatic cells around the central vein was increased to the peak(20. 60 ± 0. 47), and there was significant difference compared with other groups(all P＜0. 05). In the model groups the liver cells became edematous, and the integrity of the membrane was lost, and some cells were even lysed. The pathological damage is most serious in 24-hour model group. Conclusion The membrane attack complex C5b-9 insulted the rats' liver after a traumatic hemorrhagic shock, and apoptosis of hepatic cells and the content of C5b-9 peaked in 3-hour model, though they do not occur in the same site. A low level of C5b-9 in blood 3 hours after shock predict a poor prognosis.
