尿源性脓毒血症兔模型的建立及鲎试验的诊断价值

Establishment of rabbit urosepsis model and diagnostic value of limulus amebocyte lysate test

目的建立尿源性脓毒血症兔模型,并探讨鲎试验对尿源性脓毒血症的诊断价值。方法将18只雄性兔随机分为正常组、假手术组、脓毒血症组,每组6只。结扎脓毒血症组兔的左侧输尿管远端并向输尿管内注入大肠杆菌悬液(ATCC 25922),建立尿源性脓毒血症模型;假手术组兔仅给予暴露左侧输尿管中段,正常组不做任何处理。于术前、术后24 h、术后48 h、术后72 h,观察各组兔的肛温、呼吸频率、心率,检测外周血白细胞计数及中性粒细胞(N)计数。术后72 h,光镜观察肾组织形态结构变化,并行血培养及鲎试验。结果脓毒血症组术后的肛温、呼吸频率、心率、外周血白细胞计数及N计数均随时间延长而升高(均P <0. 05),且术后各时间点以上指标均高于正常组及假手术组(均P <0. 05),血培养为同种细菌,光镜下肾组织形态学改变明显。脓毒血症组鲎试验阳性率达100. 0%,与血培养阳性率比较差异无统计学意义(P> 0. 05)。结论将大肠杆菌悬液注入兔输尿管并结扎,72 h后可成功复制尿源性脓毒血症模型;鲎试验对尿源性脓毒血症具有早期诊断价值。

Objective To establish rabbit urosepsis model and to investigate the diagnostic value of limulus amebocyte lysate test for urosepsis. Methods Eighteen male rabbits were randomly divided into normal group,sham group and sepsis group,with 6 rabbits in each group. In the sepsis group,the left distal ureter of the rabbit was ligated then Escherichia coli suspension( ATCC 25922) was injected into the ureter to establish the urosepsis model; only exposure of medial left ureter was conducted in the sham group,no intervention was performed in the control group. Before operation and at postoperative 24,48 and 72 hours,the rectal temperature,respiratory frequency and heart rate of the rabbits were observed in each group,the counts of peripheral blood WBC and neutrophils( N) were detected. At postoperative72 hours,the morphological change of kidney tissue was observed under optical microscope,and blood culture and limulus amebocyte lysate test were performed. Results In the sepsis group,the postoperative rectal temperature,respiratory frequency,heart rate,WBC and N counts increased over time( all P < 0. 05),and were higher than those in the normal group and the sham group at each time point after operation( all P < 0. 05),the same bacteria was found in blood culture,and the obvious morphological change of kidney tissue was present under optical microscope. In the sepsis group,the positive rate of limulus amebocyte lysate test was 100. 0%,which was not statistically different from that of the blood culture( P > 0. 05). Conclusion Rabbit urosepsis model can be replicated successfully at 72 hours after injection of Escherichia coli suspension into ureter and ureter ligation. Limulus amebocyte lysate test is valuable in the early diagnosis of urosepsis.