复合感染脓毒症大鼠模型的构建和评价
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1. 云南中医药大学基础医学院,昆明 650500;2. 云南中医药大学药理教研室,昆明 650500; 3. 云南省高校中医治法与中药药效关系研究科技创新团队,昆明 650500

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Construction and evaluation of rat models of multiple-infection sepsis
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1.Department of Basic Medicine, Yunnan University of Traditional Chinese Medicine, Kunming 650500, China. 2. Department of Pharmacology, Yunnan University of Traditional Chinese Medicine, Kunming 650500. 3. Yunnan Provincial University Traditional Chinese Medicine Treatment and Traditional Chinese Medicine Efficacy Relationship Research Scientific and Technological Innovation Team, Kunming 650500

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    摘要:

    目的 着力于解决盲肠穿刺结扎(CLP)模型因水肿肠道封堵结扎穿刺部位随机影响模型动物感染进程的问题,探索通过复合感染支架整合盲肠结扎穿刺法和植入带菌物法,建立感染可持续的脓毒症动物模型。 方法 将 54 只雄性 SD 大鼠随机分为假手术组(Sham 组,n= 18)、盲肠穿刺结扎组(CLP 组,n= 18)、复合感染脓毒症模型组(MIM 组,n= 18),其中每组 10 只用于考察各组动物一般生存状态、盲肠结扎部位封堵情况和生存率; 每组另 8 只用于考察各组脓毒症损伤指标。 采用 HE 染色观察大鼠心脏、肝、肺、肾组织病理形态改变;取腹主动脉血检测脂多糖(LPS)、白介素-6( IL-6)、心肌钙蛋白-Ⅰ( cTn-I)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶 (AST)、总胆红素(TBil)、乳酸(Lac)和肌酐(Cr)含量;取肝组织检测丙二醛(MDA)和腺嘌呤核苷三磷酸(ATP)含量。 结果 造模 96 h 后,Sham 组大鼠无死亡,CLP 组大鼠生存率 70%,MIM 组生存率 0%;MIM 组心脏、肝、肺、肾 组织损伤程度较 CLP 模型组更为严重;MIM 组分别与 Sham 组和 CLP 组比,LPS、IL-6、cTn-I、ALT、AST、TBil、Cr、 Lac、MDA 含量最高,ATP 含量最低,均具有显著性差异(P< 0. 05);MIM 组 LPS、IL-6、TBil、MDA 等指标的变异系 数较 CLP 组明显更小。 结论 MIM 组可使肠内容物不断渗漏引发持续感染,反应感染和炎症的关键指标(LPS、IL- 6、MDA 等)变异性远低于 CLP 组,病程进展更为一致,其模型的结构效度更好;MIM 组较 CLP 组,术后感染表现更 为剧烈、器官损伤程度更深、而死亡率更高且组内死亡时间更为均一,其模型的表面效度更好,因而更符合严重感 染导致机体过度反应而产生脓毒症 MODS 的临床实际。

    Abstract:

    Objective We focused on solving a problem in the infection course of cecum ligation and puncture (CLP) septic model animals, i. e. , the random occlusion of ligation and puncture sites due to bowel edema. To establish a septic animal model with a sustainable infection, we combined cecal ligation puncture and bacterial carrier implantation with a compound infection stent. Methods Fifty-four male SD rats were randomly divided into three groups: Sham ( n= 18), CLP (n= 18), and multiple infect model (MIM,n= 18) groups. In each group, 10 rats were investigated for their survival status, intra-abdominal infection, and survival rate. Another 8 rats were selected for studying related indicators of sepsis multiple organ dysfunction syndrome (MODS) injury. The detected indicators included LPS, IL-6, cTn-I, ALT, AST, TBil, Lac, and Cr abdominal aortic blood contents; MDA and ATP liver tissue contents, and HE staining of pathological changes in heart, liver, lung, and kidney tissue. Results 96 h after molding operation, there were no deaths in the Sham group, and survival rates in the CLP and MIM groups were 70% and 0%, respectively. The degrees of injury in the heart, liver, lungs, and kidneys in the MIM group were greater than those in the CLP model group. Compared with the Sham and CLP groups, the MIM group had the highest contents of LPS, IL-6, cTn-I, ALT, AST, TBil, Cr, Lac, and MDA and the lowest of ATP, all of which were significantly different (P< 0. 05). The coefficients of variation for LPS, IL-6, TBil, and MDA in the MIM group were significantly decreased in relation to those of the CLP group. Conclusions The MIM group’s intestinal contents leaked continuously to cause persistent infection, which result ed in much lower variability in key infection and inflammation indicators (LPS, IL-6, MDA, etc. ) and a more consistent course of disease than in the CLP group. Thus, the structural validity of the MIM model was better. Compared with the CLP group, the MIM group had more severe postoperative infections, deeper organ damage, higher mortality, more consistent within-group death times, and thus had greater face validity. Therefore, the MIM model best fits with the clinical profile of septic-MODS, which is caused by an over-reaction to severe infection.

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段爽,朱瑛,赵茹茹,侯媛璐,李明泓,林青,李奇峰.复合感染脓毒症大鼠模型的构建和评价[J].中国实验动物学报,2022,30(8):1034~1041.

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  • 收稿日期:2022-05-10
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  • 在线发布日期: 2023-03-24
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