脂多糖注射及盲肠结扎穿刺诱导脓毒血症肾损伤模型的对比研究  

Comparative study of animal models of sepsis associated acute kidney injury induced by CLP and LPS injection

作　　者：李超[1] 石炳毅[1] 柏宏伟[1] 谢晓媚 毛文娟 刘炎忠 解俊杰[1] 崔馨芳 王爽[1] 药晨 LI Chao;SHI Bingyi;BAI Hongwei;XIE Xiaomei;MAO Wenjuan;LIU Yanzhong;XIE Junjie;CUI Xinfang;WANG Shuang;YAO Chen(The Eighth Medical Center of the General Hospital of the PLA, Beijing 100091,China;Binzhou Medical College;Peking University International Hospital;Animal Health Supervision Institute of Chengguan District,Lanzhou City)

机构地区：[1]解放军总医院第八医学中心,北京100091 [2]滨州医学院 [3]北京大学国际医院 [4]兰州市城关区动物卫生监督所

出　　处：《西北国防医学杂志》2021年第4期215-220,共6页Medical Journal of National Defending Forces in Northwest China

基　　金：中国博士后科学基金资助项目(2017T100802)。

摘　　要：目的:对比脂多糖(lipopolysaccharide,LPS)腹腔注射及盲肠结扎穿刺(cecum ligation and puncture,CLP)诱导脓毒血症肾损伤建模方法的肾脏损伤情况及炎症过程特点。方法:将48只8~10周龄C57BL/6雄性小鼠分为LPS组(n=16),磷酸盐缓冲溶液(phosphate buffer saline,PBS)组(n=8),CLP组(n=16)和假手术组(sham组,n=8)。LPS组根据小鼠体质量注射LPS溶液20 mg/kg(浓度1 mg/ml),PBS组注射等量PBS,CLP组在距结肠末端1 cm行结扎术,并用21号针对穿两孔后关闭腹腔,sham组仅开腹暴露盲肠闭合腹腔。分别在建模后0 h、6 h、12 h、24 h、48 h处死小鼠,比较观察期内各组小鼠肾功能指标、病理损伤及炎症指标变化。用免疫荧光染色观察肾小管上皮细胞的凋亡。结果:PBS组及sham组小鼠建模前后血清肌酐、尿素氮水平比较,差异无统计学意义(P>0.05);而LPS组和CLP组小鼠肌酐、尿素氮迅速升高,均在24 h达峰值,分别与建模前比较,差异有统计学意义(均P<0.05),且48 h后均逐渐恢复。肾脏病理检查提示,LPS组及CLP组均显示肾小管上皮细胞空泡形成明显增多,小管坏死及凋亡不明显,但炎症反应过程有显著差别,其中LPS组白介素6、肿瘤坏死因子等炎症因子水平较CLP组迅速升高,6 h达到峰值后逐渐降低,且CLP组下降较LPS组缓慢。细胞凋亡染色显示LPS组和CLP组均存在局部凋亡,而PBS组和sham组均未见明显凋亡。结论:LPS腹腔注射及CLP均能成功建立脓毒血症肾损伤模型,其中LPS模型的全身性炎症反应剧烈,重复性好,而CLP模型更贴近临床败血症的炎症特点,在科研实践中可根据需要选择合适的模型。Objective:To compare the renal injury and inflammatory characteristics of sepsis associated acute kidney injury(SA-AKI)induced by lipopolysaccharide(LPS)injection and cecum ligation puncture(CLP).Methods:Forty-eight male mice C57BL/6 aged 8-10 weeks were divided into the LPS group(n=16)and the phosphate buffer saline(PBS)group(n=8),the CLP group(n=16)and sham group(n=8).LPS group was injected with LPS solution of 20 mg/kg(with the concentration of 1 mg/ml)according to the weight,PBS group was injected with the same amount of PBS,and CLP group performed ligation at a distance of 1 cm from the end of colon,and the peritoneal cavity was closed with a 21-gauge needle,sham group only exposed cecum and where after closed the peritoneal cavity without cecal ligation and puncture.The mice were sacrificed at 0 h,1 h,12 h,24 h,48 h after establishing model,and renal function indexes,pathological injury and inflammation in each group was compared during the observation period.Immunofluorescence staining was used to observe the apoptosis of renal tubular epithelial cells.Results:There was no significant difference in serum creatinine(SCr)and blood urea nitrogen(BUN)levels between PBS group and sham group before and after modeling(P>0.05).BUN and SCr in the LPS group and the CLP group increased and peaked at 24 hours,the difference was statistically significant compared with that before modeling(P<0.05),and mice recoverd gradually after 48 h.Pathological examination showed that vacuoles of renal tubular epithelial cells increased significantly in the LPS group and the CLP group,while tubular necrosis and apoptosis were not obvious in the two groups.There were significant differences in the inflammatory response process between the LPS group and the CLP group,interleukin-6 and tumor necrosis factor-αincreased and peaked at 6 hours,and then decreased gradually,the CLP group decreased more slowly than the LPS group.Apotosis staining showed focal apoptosis exisisted in LPS group and CLP group,while no obvious apoptosis was found

关 键 词：急性肾损伤 雄性小鼠 脓毒血症 脂多糖 盲肠结扎穿刺 炎症因子 

分 类 号：R692[医药卫生—泌尿科学]