【摘要】 目的研究高迁移率族蛋白B1(high mobility group box-1 protein,HMGB1)mRNA表达对急性坏死性胰腺炎(acute necrotizing pancreatitis,ANP)大鼠肠黏膜紧密连接蛋白-1(zonula occludens protein-1,ZO-1)表达的影响,初步探讨ANP时肠黏膜屏障损伤的可能机理。方法将96只Wistar大鼠随机(随机数字表法)均分为ANP组、丙酮酸乙酯(EP)组及假手术组,3组均分别于术后6、12、24及48 h时各抽取8只大鼠,取腹主动脉血和回肠组织。采用全自动生化分析仪检测血淀粉酶(AMY)水平,采用改良酶学分光光度法检测血浆D-乳酸水平,采用硫代巴比妥酸(TAB)比色法检测回肠组织丙二醛(MDA)含量,采用HE染色法观察大鼠回肠组织的病理学改变,采用免疫组织化学法(SP法)观察回肠组织ZO-1蛋白的表达,采用逆转录聚合酶链反应(RT-PCR)法检测回肠组织HMGB1和ZO-1 mRNA的表达。结果各时相EP组大鼠的血AMY、D-乳酸和回肠组织MDA含量均低于ANP组(P<0.05)。6 h时,ANP组大鼠回肠组织HMGB1 mRNA的表达即上调,但ZO-1 mRNA的表达下调;各时相EP组大鼠回肠组织HMGB1 mRNA的表达水平均低于ANP组,但ZO-1 mRNA的表达水平均高于ANP组(P<0.05)。各时相EP组大鼠回肠组织的病理学损伤均较ANP组明显减轻。结论 ANP大鼠回肠黏膜组织中ZO-1表达下调是ANP大鼠肠黏膜屏障损伤的重要原因之一,可能与炎症介质HMGB1的过度表达有关。更多还原

【Abstract】 Objective To study the effect on expression of high mobility group box-1(HMGB1)mRNA for the expression of zonula occludens-1(ZO-1)in ileum tissues,and to explore the possible mechanism of intestinal mucosal barrier injury in rats with acute necrotizing pancreatitis(ANP).Methods Ninety-six male Wistar rats were divided randomly(random number method)into ANP group,ethyl pyruvate(EP)group,and sham operation group.Eight rats of 3 groups were killed to get abdominal aortic blood and ileal tissues at 6,12,24,and 48 h after operation,respectively.The levels of plasma amylase(AMY),D-lactate acid,and the activity of malonyl dialdehyde(MDA)in the ileum tissues were determined by using automatic biochemical analyzer,improved enzymatic spectrophotometry,and thiobarbituric acid(TAB)colorimetry respectively.The pathological changes of ileum tissues were observed under microscopy by HE staining,the expression of ZO-1 protein in ileum tissues was observed by immunohistochemistry(SP method),and the expressions of HMGB1 mRNA and ZO-1 mRNA in ileum tissues were detected by reverse transcription-polymerase chain reaction(RT-PCR).Results Compared with ANP group at the same time,levels of AMY,D-lactate acid,and MDA in ileum tissues of EP group were all significantly lower(P<0.05).The expression level of HMGB1 mRNA increased at 6 h while ZO-1 mRNA decreased in ANP group.Compared with ANP group at the same time,the expression level of HMGB1 mRNA of EP group was significantly lower while ZO-1 mRNA was higher(P < 0.05),and the pathological damage in ileum tissues was lighter.Conclusions The decreased expression of ZO-1 in ileum tissues is one of the vital causes for intestinal mucosal barrier injury in ANP,and it probably occurs in case of the excessive expression of HMGB1.更多还原