质粒载体示踪法用于急性坏死性胰腺炎肠道细菌易位的研究

An experimental study on intestinal bacterial translocation with JM109 bearing plasmid PUC18 in acute necrotizing pancreatitis

目的：研究急性坏死性胰腺炎（ＡＮＰ）时肠道细菌易位情况，探讨肠道是否为继发性胰腺感染的细菌来源。方法：采用ＰＵＣ１８质粒载体大肠肝菌（ＪＭ１０９）示踪法，将质粒菌ＪＭ１０９定植于１５只犬肠道后，随机分为ＡＮＰ组（８只）及对照组（７只），前者复制ＡＮＰ模型。每日进行血培养，第７日活杀后作脏器细菌培养。对耐氨苄青霉素菌株进行酶切分析及琼脂糖凝胶电泳。结果：胰腺炎犬除肾及脾脏外，其余脏器均发现有ＪＭ１０９菌易位，以胰腺易位率最高（７５％，６／８）；发病后２４小时血培养中找到质粒菌ＪＭ１０９阳性率高达５０％。结论：ＡＮＰ发病后２４小时即可出现细菌易位。

Objective:To observe intestinal bacterial translocation in acute necrotizing pancreatitis (ANP),and to elucidate whether the gut would be the source of bacteria in pancreatic infection.Methods:In this study,JM109 labeled plasmid PUC 18 was given by gavage to detect intestinal bacterial translocation in ANP.Fifteen dogs were colonized with a strain of E.coli JM 109 labeled ampicilllin[CD*2]resistant plasmid PUC18,and then divided into 2 groups.In ANP group ( n =8),ANP was induced by injection of 0 5 ml/kg of sodium taurocholate with 3 000 U/kg trypsin into the pancreatic duct.The control group ( n =7) underwent laparotomy only.Blood was cultured every day,all animals were sacrificed on postoperative day 7 and organs were harvested for culture.Ampicillin[CD*2]resistant strains with JM 109 bearing plasmid PUC 18,which was colonized to the animals previously,were identified by restriction digest and agarose gel[CD*2]electrophoresis.Results:JM 109 was detected in mesenteric lymph nodes,pancreas,liver,lungs and blood of ANP animals.The isolation rates in the pancreas and blood were 75% and 50%,respectively.Conclusions:Bacterial translocation from the gut could taked place early after the onset of ANP,and serve as a potential source of secondary pancreatic infection.