肠道菌群对急性坏死性胰腺炎小鼠肺损伤的影响  

Effect of intestinal microbiota on lung injury in mice with acute necrotizing pancreatitis

作　　者：赵梦琦 崔梦妍 范妙言 陆颖影 江巧丽 Zhao Mengqi;Cui Mengyan;Fan Miaoyan;Lu Yingying;Jiang Qiaoli(Department of Gastroenterology,Jiading Branch of Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 201803,China;Department of Gastroenterology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 201620,China)

机构地区：[1]上海交通大学医学院附属第一人民医院嘉定医院消化内科,上海201803 [2]上海交通大学医学院附属第一人民医院消化内科,上海201602

出　　处：《中华胰腺病杂志》2025年第2期126-133,共8页Chinese Journal of Pancreatology

基　　金：上海市嘉定区自然科学研究课题(JDKW-2022-0042);上海市嘉定区卫生健康委员会科研课题(2022-KY-01)。

摘　　要：目的探讨肠道菌群对急性坏死性胰腺炎(ANP)小鼠肺损伤的影响及其机制。方法小鼠随机分为正常对照组(CON组)、ANP模型组(ANP组)、肠道无菌组(ABX组),每组6只。采用腹腔注射雨蛙肽(100μg/kg)10次,每次间隔1 h,随后再注射脂多糖(10 mg/kg)构建ANP小鼠模型;ABX组小鼠应用Abx溶液(0.5 g/L万古霉素、1 g/L新霉素、1 g/L甲硝唑和1 g/L氨苄西林)1 ml/100 g灌胃28 d后再制备ANP模型;CON组腹腔注射等体积磷酸盐缓冲溶液。常规行胰腺、肺和回肠组织病理学检查。采用酶化学动力法检测血清淀粉酶水平,ELISA法检测血清二胺氧化酶(DAO)和肺组织髓过氧化物酶(MPO)活性;荧光定量PCR法检测肺组织炎症因子、细胞焦亡相关分子及回肠上皮紧密连接蛋白mRNA的表达;蛋白质印迹法检测肺组织细胞焦亡分子蛋白及回肠上皮紧密连接蛋白的表达;荧光原位杂交技术检测肺组织细菌移位情况。结果CON组、ANP组、ABX组胰腺组织病理评分分别为(0.67±0.26)、(7.33±0.82)、(5.67±0.81)分,肺组织分别为(1.67±0.41)、(5.67±0.41)、(3.58±0.58)分,回肠组织分别为(0.58±0.52)、(3.83±0.75)、(4.33±0.82)分;血清淀粉酶水平分别为(403.95±93.11)、(1037.24±126.77)、(647.32±145.90)U/L,肺组织MPO水平分别为(0.23±0.03)、(0.63±0.09)、(0.48±0.05)U/g。ABX组胰腺组织及肺组织病理评分、血清淀粉酶、肺组织MPO均较ANP组显著降低,差异有统计学意义(P值均<0.05)。CON组、ANP组、ABX组胰腺组织IL-1βmRNA表达量分别为1.84±0.90、36.26±5.56、16.65±6.43,IL-6 mRNA表达量分别为1.07±0.15、2.90±0.42、1.34±0.62,TNF-αmRNA表达量分别为0.47±0.11、0.76±0.11、0.46±0.07,HMGB1 mRNA表达量分别为0.38±0.02、0.72±0.22、0.44±0.08,caspase-1 mRNA表达量分别为1.07±0.18、2.04±0.31、0.85±0.54,ASC mRNA表达量分别为1.24±0.19、5.68±0.41、3.89±1.47,GSDMD mRNA表达量分别为0.79±0.17、0.94±0.14、0.61±0.08,IL-18 mRNA表达量分别�ObjectiveTo explore the effects and possible mechanism of intestinal microbiota on lung injury in mice with acute necrotizing pancreatitis(ANP).MethodsThe experimental mice were randomly assigned to normal control group(CON group),ANP model group(ANP group)and intestinal germ-free group(ABX group),with 6 mice in each group.The ANP mouse model was constructed by intraperitoneal injection of caerulein(100μg/kg,for 10 times)at an interval of 1 hour each time,followed by 10 mg/kg lipopolysaccharide injection.Mice in ABX group were treated by Abx solution(0.5 g/L vancomycin,1 g/L neomycin,1 g/L metronidazole,and 1 g/L ampicillin),1 ml/100 g gavage for 28 days before preparation of the ANP model.The CON group was injected intraperitoneally with an equal volume of PBS.Histopathologic examination of the pancreas,lungs,and terminal ileum was routinely performed.Serum amylase levels were measured using enzymatic kinetic chemistry,and serum diamine oxidase(DAO)and lung tissue myeloperoxidase(MPO)activities were measured using ELISA assay.Expression of inflammatory factors,pyroptosis-related molecules in lung tissue and intestinal epithelial tight junction proteins was detected by fluorescence quantitative PCR.Western blotting was used to detect the expression of the pyroptosis molecules caspase-1 and GSDMD in lung tissue,and intestinal epithelial tight junction proteins.Changes of bacterial distribution in lung tissue were measured by fluorescence in situ hybridization.ResultsThe pathological scores of pancreatic tissue of CON,ANP,and ABX group were(0.67±0.26),(7.33±0.82),and(5.67±0.81);the pathological scores of lung tissue were(1.67±0.41),(5.67±0.41),and(3.58±0.58);the pathological scores of ileal tissue were(0.58±0.52),(3.83±0.75),and(4.33±0.82);the serum amylase levels were(403.95±93.11),(1037.24±126.77),and(647.32±145.90)U/L;the MPO levels in lung tissue were(0.23±0.03),(0.63±0.09),and(0.48±0.05)U/g.ABX group had significantly lower scores in pancreatic and lung tissues,serum amylase levels,and MPO leve

关 键 词：胰腺炎 急性坏死性 肺损伤 焦亡 肠道菌群 

分 类 号：R576[医药卫生—消化系统]