构建大鼠脑出血致多器官功能障碍综合征模型(英文)

Construction of rat model of multi-organ dysfunction syndromes induced by cerebral hemorrhage

背景:急性脑血管病致多器官功能障碍综合征多见于临床资料分析,有关急性脑血管病后的周围器官功能障碍的实验甚为匮乏。目的:采用两种剂量的胶原酶加上适量肝素复制脑出血致多器官功能障碍综合征的动物模型,摸索最佳实验条件。设计:完全随机对照实验。单位:山东大学齐鲁医院的皮肤科、神经内科、心外科和济南铁路中心医院的神经内科。材料:实验于2002-09/12在山东省医学科学院基础医学研究所进行。96只成年健康雄性Wistar大鼠随机分为正常对照组6只,假手术组6只,出血1组42只,出血2组42只,出血1组,出血2组又分为4,8,12,24,36,48,72h的7个亚组,每个亚组6只。干预:出血1组脑内缓慢注入Ⅶ型胶原酶0.4U和3.2IU的肝素钠混合液2μL,出血2组脑内注入Ⅶ型胶原酶0.8U和3.2IU的肝素钠混合液2μL,假手术组注射同体积的生理盐水。观察术后各时相点动物的一般情况,包括意识、精神状态、肢体活动,体温、呼吸、心率改变;检测血常规、肝功能(丙氨酸氨基转移酶,天冬氨酸氨基转移酶)、肾功能(尿素氮、肌酐)、心肌酶学(肌酸激酶,乳酸脱氢酶)。采用改良基质显色法鲎试验测定血浆内毒素含量。光镜下观察脑、肺脏、肝脏、小肠、肾脏的病理改变。主要观察指标:①术后实验动物的一般情况。②各组动物的肝功能、肾功能、心肌酶学、血浆内毒素含量的测定结果。③实验动物的主要脏器病理改变。结果:96只大鼠全部进入结果分析。①术后实验动物的一般情况:出血1组、2组动物的体温、呼吸、心率、白细胞含量明显高于正常组、假手术组(P<0.05)。出血2组高于出血1组犤(39.56±0.59)℃,(37.98±0.52)℃;(124.23±4.12)次/min,(92.56±2.95)次/min;(325.96±84.53)次/min,(258.16±75.42)次/min,(P<0.05)犦,尤其以外周血白细胞含量差异明显犤(19.46±0.91)×109/L,(10.83±0.87)×109/L,(P<0.01)犦。②各组动物的静脉血中丙氨酸氨基转移酶活性、天冬氨酸氨基转移酶活性、尿素氮含量、肌酐含量、肌酸激酶活性、乳酸脱氢酶活性、血浆内毒素含量的测定结果:出血1组、2组高于正常组和假手术组(P<0.05～0.01〉,出血2组高于出血1组犤(35.04±7.54)μkat/L,(24.25±5.42)μkat/L;(51.61±10.47)μkat/L,(33.77±7.44)μkat/L;(21.12±2.93)mmol/L,(9.78±1.45)mmol/L;(68.57±5.24)μmol/L,(40.54±3.95)μmol/L;(706.44±44.92)μkat/L,(189.23±25.08)μkat/L;(357.96±40.95)μkat/L,(149.60±19.10)(P<0.05)犦,尤其以血浆内毒素含量差异明显犤(1.025±0.250)Eu/mL,(0.595±0.150)Eu/mL,(P<0.01)犦。③实验动物的主要脏器病理改变:大鼠脑出血各时相点动物的器官组织均有不同程度的炎性损害,出血1组在24～36h的脏器病理变化达到高峰,72h基本恢复正常,出血2组较出血1组炎性损害更加明显持久,在24～48h的脏器病理变化达到高峰,72h仍可见炎性损害。结论:显示0.8U胶原酶模型是较成功的脑出血致多器官功能障碍综合征模型。该模型符合临床常见诱发因素,各器官功能的各项指标变化能动态反应多器官功能障碍综合征的发病过程。

BACKGROUND： There is much clinical datum analysis on multi-organ dysfunction syndromes induced by cerebral hemorrhage, but there is lack of experiment on dysfunction of peripheral organs induced by acute cerebral vascular disease. OBJECTIVE： Collagenase of two dosages together with proper amount of heparin were used to duplicate animal model of multi-organ dysfunction syndromes induced by cerebral hemorrhage so as to probe into the best experimental prerequisite. DESIGN： Complete randomized control experiment. SETTING： Departments of Dermatology, Neurology and Cardiac Surgery of Qilu Hospital Affiliated to Shandong University and Department of Neurology of Jinan Railway Center Hospital. MATERIALS： The experiment was performed in Institute of Basic Medicine of Shandong Medical Scientific Academy from September to December 2002. Totally 96 adult healthy male Wistar rats were employed and randomized into normal control （6 rats）, sham-operation group （6 rats）, first hemorrhage group （42 rats） and second hemorrhage group （42 rats）. Seven sub-groups of 4 hours, 8 hours, 12 hours, 24 hours, 36 hours, 48 hours and 72 hours successively were divided in first and second hemorrhage groups respectively, 6 rats in each sub-group. INTERVENTIONS： In the first hemorrhage group, mixed solution 2μL of VR collagenase 0.4 U and heparin sodium 3.2IU was infused slowly to the brain. In the second hemorrhage group, mixed solution 2μL of VR collagenase 0.8 U and heparin sodium 3.2 IU was infused slowly to the brain. In sham-operation group, physiological saline of same volume was injected. The general situation of animals at every phasic point was observed after operation, including consciousness, mental state and limb movement, as well as the ahemations on body temperature, respiration and heart rate. Examinations were done on blood routine, liver function （alanine aminopherase, ALT; aspartic acid translocase, AST）, kidney function （urea nitrogen, BUN; creatinine CRE） and mycs＇ardial zymology （creatine kinase, CK; lactic dehydrogenase, LDH）. Modified matrix coloration method was used to determine endotoxin content in plasma. Pathological alternations were observed optic microscopically on brain, lung, liver, small intestine and kidney. MAIN OUTCOME MEASURES： ① General situation of experimental animals after operation. ② Determination of liver function, kidney function myocardial zymology and endotoxin content in plasma. ③ Pathological alternations on main organs of experimental animals. RESULTS： All of 96 rats entered result analysis. ① General situation of experimental animals after operation： In the first and second hemorrhage groups, body temperature, respiration, heart rate and leukocyte content were all higher remarkably than normal group and sham-operation group （P 〈 0.05）.Those in the first hemorrhage group were higher than second group [（39.56±0.59）℃, （37.98±0.52）℃;（124.23±4.12） timeJminute, （92.56±2.95） times/minute; （325.96±84.53） times/minute, （258.16±75.42） times/minute, （P 〈 0.05）], and the difference in peripheral leukocyte content was specially obvious [（19.46±0.91）×10^9/L, （10.83±0.87）×10^9/L,（P 〈 0.01）]. ② Determination of ALT activity, AST activity, BUN content, CRE content, CK activity, LDH activity and endotoxin content in plasma： Those in the first and second hemorrhage groups were higher than normal group and sham-operation group （P 〈 0.05 -0.01）. Those in the first hemorrhage group were higher than second hemorrhage group [（35.04±7.54）μkat /L,（24.25±5.42）μkat/L; （51.61±10.47） μkat/L, （33.77±7.44） μkat/1,; （21.12±2.93） mmol/L,（9.78±1.45） mmol/L; （68.57±5.24） μmol/L （40.54±3.95） μmol/L; （706.44 ±4.92） μkat/L, （189.23±25.08） μkat/L; （357.96±40.95） μkat/L, （149.60±19.10） （P 〈 0.05）] and the difference was especially remarkable in endotoxin content in plasma [（1.025±0.250） Eu/mL, （0.595±0.150） Eu/mL, （P 〈 0.01）]. ③Pathological alternations on main organs of experimental animals： Inflammatory damage of various degrees presented in all of animal organ tissues at various phasic points during cerebral hemorrhage in rats. In the first hemorrhage group, pathological change of organs reached the peak in 24-36 hours, which recovered to be normal in 72 hours. In the second hemorrhage group, the inflammatory damage was more remarkable and persistent compared with first group, in which, pathological change of organs reached the peak in 24-48 hours and inflammatory damage was still visible in 72 hours. CONCLUSION： It is indicated that 0.8 U collagenase model is a successful model of multi-organ dysfunction syndromes induced by cerebral hemorrhage. Such model is in conformity with the common induced factors in clinic and various index changes of functions in every organ and can reflect dynamically the occurrence of multi-organ dysfunction syndromes.