复合感染脓毒症大鼠模型的构建和评价_段爽.pdf

2022 年 12 月第 30 卷 第 8 期中国实验动物学报ACTA LABORATORIUM ANIMALIS SCIENTIA SINICADecember 2022Vol.30 No.8段爽,朱瑛,赵茹茹,等.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.Duan S,Zhu Y,Zhao RR,et al.Construction and evaluation of rat models of multiple-infection sepsis J.Acta Lab Anim Sci Sin,2022,30(8):1034-1041.Doi:10.3969/j.issn.1005-4847.2022.08.003基金项目国家自然科学基金(81960815),云南省科学技术厅联合专项(202001A2070001-032),云南中医药大学南药可持续利用研究重点实验室(202105AG070012XS2245)。Funded by the National Natural Science Foundation of China(81960815),Joint Special Project of Yunnan Provincial Department of Science and Technology(202001A2070001-032),Key Laboratory of Sustainable Utilization of Southern Medicine of Yunnan University of Chinese Medicine(202105AG070012XS2245).作者简介段爽(1996),女,在读硕士研究生,研究方向:中西医结合防治疾病作用机制研究。Email:944765534 通信作者李奇峰(1977),男,硕士,副教授,研究方向:中西医临床基础。Email:375487824 复合感染脓毒症大鼠模型的构建和评价段爽1,朱瑛1,赵茹茹1,侯媛璐1,李明泓1,3,林青2,3,李奇峰1,3(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)【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-(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。【关键词】脓毒症;多器官功能障碍综合征;复合感染模型;盲肠结扎穿刺【中图分类号】Q95-33 【文献标识码】A 【文章编号】1005-4847(2022)08-1034-08Construction and evaluation of rat models of multiple-infection sepsisDUAN Shuang1,ZHU Ying1,ZHAO Ruru1,HOU Yuanlu1,LI Minghong1,3,LIN Qing2,3,LI Qifeng1,3(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)Corresponding author:LI Qifeng.E-mail:375487824 【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 中国实验动物学报 2022 年 12 月第 30 卷第 8 期 Acta Lab Anim Sci Sin,December 2022,Vol.30,No.8with a compound infection stent.MethodsFifty-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 groups 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.【Keywords】sepsis;multiple-organ dysfunction syndrome;multiple-infection model;cecum ligation punctureConflicts of Interest:The authors declare no conflict of interest.脓毒症是由于严重感染引起机体过度反应而出现危机生命的多器官功能障碍综合征(multiple organ dysfunction synd