大鼠盲肠结扎穿孔脓毒症模型建模新方案的应用研究
发布时间：2018-11-18 08:12
【摘要】：目的:目前模拟临床脓毒症最佳的动物模型为啮齿类动物的盲肠结扎穿孔(CLP)模型,其主要方法是分别通过控制盲肠穿刺针粗细、穿孔个数和盲肠结扎的长度来控制实验动物的死亡率,但在实际应用过程中其实验结果可重复性较差。本研究旨在建立以标测盲肠体积为基础的CLP模型来进一步控制动物感染严重程度与死亡率,同时模拟以往文献中以标测盲肠长度为基础的CLP模型进行对比,探索提高模型稳定性的可行性方案。 方法:采用雄性SD大鼠建立盲肠结扎穿孔模型,分长度标测结扎及体积标测结扎两大组,长度标测组用丝线测量由回盲瓣至盲肠顶端全长,按不同长度比例分亚组结扎盲肠(盲肠全长的50%及75%),体积标测组用亚甲蓝标记盲肠体积,按不同体积分亚组结扎盲肠(0.5ml,1.0ml, 1.5ml, 2.0ml和2.5ml);结扎后均用21号针头单次贯通穿孔盲肠,术后6小时进行血培养及腹腔分泌物涂片培养并观察7天死亡率。 结果:长度标测结扎组内50%和75%组7天死亡率分别为90%和100%,两组间无显著差异(n=10, p =0.661)。体积标测结扎组内死亡率随盲肠结扎体积增加而上升,0.5ml,1.0ml,1.5ml,2.0ml和2.5ml各组7天死亡率分别为40%、50%、60%、80%和100%,各组间存在显著差异(n=10, p =0.002),而假手术组全部存活。长度标测结扎组50%和75%亚组平均存活时间分别为1.621±0.605天和1.150±0.099天。体积标测结扎组平均存活时间从0.5ml结扎亚组的4.708±0.893天到2.5ml结扎亚组的1.175±0.240天。CLP术后6小时血培养及腹腔分泌物涂片培养除假手术组外均为阳性。 结论:手术操作过程及实验数据显示盲肠体积结扎穿孔法是一项更容易进行的标准化操作,是实验结果重复性更好的CLP造模方法,可推广应用于对脓毒症机制的研究以及对药物疗效进行评估。
[Abstract]:Objective: the best animal model to simulate clinical sepsis is the (CLP) model of rodent cecal ligation and perforation. The main method is to control the thickness of cecum puncture needle. The number of perforations and the length of cecal ligation were used to control the mortality of experimental animals. The purpose of this study was to establish a CLP model based on measuring the volume of cecum to further control the severity and mortality of animal infection, and to simulate the CLP model based on measuring the length of cecum in previous literatures for comparison. To explore a feasible scheme to improve the stability of the model. Methods: the model of cecal ligation and perforation was established in male SD rats. The model was divided into two groups: length mapping ligation and volume mapping ligation. The length of length from ileocecal flap to cecum apical was measured by silk line in length mapping group. Cecum was ligated by different length ratio (50% and 75% of the total length of cecum). The volume of cecum was labeled with methylene blue in the volume mapping group, and the cecum was ligated in different volume groups (0.5 ml, 1.5 ml, 2.0ml and 2.5ml). After ligation, the perforated caecum was single perforated with needle 21. The blood culture and peritoneal secretion smear culture were performed 6 hours after the operation and the death rate was observed for 7 days. Results: the 7-day mortality of 50% and 75% of the ligation group was 90% and 100%, respectively. There was no significant difference between the two groups (n = 10, p = 0.661). The mortality in the group of volume mapping and ligation increased with the increase of the volume of the cecal ligation. The mortality rates of 0.5 ml of 1.0 ml of 1. 0 ml 1. 5 ml of 1. 5 ml and 2. 0 ml of 2.5ml for 7 days were 40% and 50%, 80% and 100%, respectively. There was a significant difference among the three groups (n = 10, p = 0.002), but all of the sham operation groups survived. The mean survival time of 50% and 75% subgroups was 1.621 卤0.605 days and 1.150 卤0.099 days, respectively. The mean survival time in the volume mapping ligation group ranged from 4.708 卤0.893 days in the 0.5ml ligation subgroup to 1.175 卤0.240 days in the 2.5ml ligation subgroup. 6 hours after CLP, blood culture and peritoneal secretion smear culture were positive except the sham operation group. Conclusion: the operative procedure and experimental data show that the method of volume ligation and perforation of the caecum is a more convenient and standardized procedure, and it is a more reproducible CLP model method. It can be used to study the mechanism of sepsis and evaluate the efficacy of drugs.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R459.7;R-332
[Abstract]:Objective: the best animal model to simulate clinical sepsis is the (CLP) model of rodent cecal ligation and perforation. The main method is to control the thickness of cecum puncture needle. The number of perforations and the length of cecal ligation were used to control the mortality of experimental animals. The purpose of this study was to establish a CLP model based on measuring the volume of cecum to further control the severity and mortality of animal infection, and to simulate the CLP model based on measuring the length of cecum in previous literatures for comparison. To explore a feasible scheme to improve the stability of the model. Methods: the model of cecal ligation and perforation was established in male SD rats. The model was divided into two groups: length mapping ligation and volume mapping ligation. The length of length from ileocecal flap to cecum apical was measured by silk line in length mapping group. Cecum was ligated by different length ratio (50% and 75% of the total length of cecum). The volume of cecum was labeled with methylene blue in the volume mapping group, and the cecum was ligated in different volume groups (0.5 ml, 1.5 ml, 2.0ml and 2.5ml). After ligation, the perforated caecum was single perforated with needle 21. The blood culture and peritoneal secretion smear culture were performed 6 hours after the operation and the death rate was observed for 7 days. Results: the 7-day mortality of 50% and 75% of the ligation group was 90% and 100%, respectively. There was no significant difference between the two groups (n = 10, p = 0.661). The mortality in the group of volume mapping and ligation increased with the increase of the volume of the cecal ligation. The mortality rates of 0.5 ml of 1.0 ml of 1. 0 ml 1. 5 ml of 1. 5 ml and 2. 0 ml of 2.5ml for 7 days were 40% and 50%, 80% and 100%, respectively. There was a significant difference among the three groups (n = 10, p = 0.002), but all of the sham operation groups survived. The mean survival time of 50% and 75% subgroups was 1.621 卤0.605 days and 1.150 卤0.099 days, respectively. The mean survival time in the volume mapping ligation group ranged from 4.708 卤0.893 days in the 0.5ml ligation subgroup to 1.175 卤0.240 days in the 2.5ml ligation subgroup. 6 hours after CLP, blood culture and peritoneal secretion smear culture were positive except the sham operation group. Conclusion: the operative procedure and experimental data show that the method of volume ligation and perforation of the caecum is a more convenient and standardized procedure, and it is a more reproducible CLP model method. It can be used to study the mechanism of sepsis and evaluate the efficacy of drugs.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R459.7;R-332
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1 郭媛;魏筱华;谢s，
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