中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
10期
1093-1098
,共6页
胰腺炎%肠屏障%高迁移率族蛋白B1%紧密连接
胰腺炎%腸屏障%高遷移率族蛋白B1%緊密連接
이선염%장병장%고천이솔족단백B1%긴밀련접
Pancreatitis%Intestinal mucosal barrier%High mobility group box-1 protein%Tight junction
目的 观察重症急性胰腺炎(SAP)大鼠肠组织中高迁移率族蛋白B1 (HMGB1)表达对肠黏膜上皮细胞紧密连接occludin蛋白表达的影响.方法 逆行胰胆管注射5％牛磺胆酸钠制作SAP模型.健康Wistar大鼠随机(随机数字法)分为对照组、SAP组、二硫代氨基甲酸吡咯烷(PDTC)处理组.测定血淀粉酶(AMY)、内毒素(LPS)及D-乳酸水平；光镜下观察胰腺和肠组织的病理变化；免疫组织化学法观察occludin分布和表达的变化；RT-PCR法检测大鼠肠组织中HMGB1的表达水平；Western blot法检测大鼠肠组织中HMGB1及occludin蛋白的表达水平.采用SPSS 13.0统计分析软件进行处理,P＜ 0.05为差异具有统计学意义.结果 在建模后24 h,SAP组大鼠血浆LPS及D-乳酸水平明显高于对照组,提示肠屏障通透性明显增加；PDTC处理组大鼠血浆LPS及D-乳酸水平明显低于SAP组(P＜0.05).SAP组大鼠肠组织HMGB1表达水平较对照组明显升高,而occludin蛋白的表达较对照组下降；PDTC组大鼠肠组织HMGB1表达水平明显低于SAP组,occludin水平较SAP组升高(P＜0.05).结论 SAP时,大鼠肠组织内HMGB1表达升高,通过降低occludin蛋白表达,来增加肠黏膜屏障通透性；PDTC可抑制HMGB1表达,上调occludin蛋白表达,改善肠黏膜屏障通透性.
目的 觀察重癥急性胰腺炎(SAP)大鼠腸組織中高遷移率族蛋白B1 (HMGB1)錶達對腸黏膜上皮細胞緊密連接occludin蛋白錶達的影響.方法 逆行胰膽管註射5％牛磺膽痠鈉製作SAP模型.健康Wistar大鼠隨機(隨機數字法)分為對照組、SAP組、二硫代氨基甲痠吡咯烷(PDTC)處理組.測定血澱粉酶(AMY)、內毒素(LPS)及D-乳痠水平；光鏡下觀察胰腺和腸組織的病理變化；免疫組織化學法觀察occludin分佈和錶達的變化；RT-PCR法檢測大鼠腸組織中HMGB1的錶達水平；Western blot法檢測大鼠腸組織中HMGB1及occludin蛋白的錶達水平.採用SPSS 13.0統計分析軟件進行處理,P＜ 0.05為差異具有統計學意義.結果 在建模後24 h,SAP組大鼠血漿LPS及D-乳痠水平明顯高于對照組,提示腸屏障通透性明顯增加；PDTC處理組大鼠血漿LPS及D-乳痠水平明顯低于SAP組(P＜0.05).SAP組大鼠腸組織HMGB1錶達水平較對照組明顯升高,而occludin蛋白的錶達較對照組下降；PDTC組大鼠腸組織HMGB1錶達水平明顯低于SAP組,occludin水平較SAP組升高(P＜0.05).結論 SAP時,大鼠腸組織內HMGB1錶達升高,通過降低occludin蛋白錶達,來增加腸黏膜屏障通透性；PDTC可抑製HMGB1錶達,上調occludin蛋白錶達,改善腸黏膜屏障通透性.
목적 관찰중증급성이선염(SAP)대서장조직중고천이솔족단백B1 (HMGB1)표체대장점막상피세포긴밀련접occludin단백표체적영향.방법 역행이담관주사5％우광담산납제작SAP모형.건강Wistar대서수궤(수궤수자법)분위대조조、SAP조、이류대안기갑산필각완(PDTC)처리조.측정혈정분매(AMY)、내독소(LPS)급D-유산수평；광경하관찰이선화장조직적병리변화；면역조직화학법관찰occludin분포화표체적변화；RT-PCR법검측대서장조직중HMGB1적표체수평；Western blot법검측대서장조직중HMGB1급occludin단백적표체수평.채용SPSS 13.0통계분석연건진행처리,P＜ 0.05위차이구유통계학의의.결과 재건모후24 h,SAP조대서혈장LPS급D-유산수평명현고우대조조,제시장병장통투성명현증가；PDTC처리조대서혈장LPS급D-유산수평명현저우SAP조(P＜0.05).SAP조대서장조직HMGB1표체수평교대조조명현승고,이occludin단백적표체교대조조하강；PDTC조대서장조직HMGB1표체수평명현저우SAP조,occludin수평교SAP조승고(P＜0.05).결론 SAP시,대서장조직내HMGB1표체승고,통과강저occludin단백표체,래증가장점막병장통투성；PDTC가억제HMGB1표체,상조occludin단백표체,개선장점막병장통투성.
Objective To observe the effect of high mobility group box-1 protein (HMGB1) on the expression of intestinal epithelial tight junction protein occludin in murine severe acute pancreatitis (SAP).Methods Rat SAP model was estabilished by retrograde injection of 5 ％ sodium taurocholate into choledochopancreatic duct.Healthy wistar rats were divided randomly (random number) into three groups:control group,SAP group,pyrrolidine dithiocarbamate (PDTC) therapy group.Levels of plasm amylase,lipopolysaccharide (LPS) and D-lactate were determined.The changes of morphological damage of pancreasand intestinal tissues were observed by microscopy.The distribution and expression of occludin protein were observed by SP immunohistochemistry. The mRNA expression of HMGB1 in the intestinal mucosa was detected by reverse-transcription polymerase chain reaction (RT-PCR).The expressions of HMGB1 and occludin were determined by western blotting. One-way analysis of variance was performed with SPSS Windows 13.0 statistical analysis software,and a difference was accepted as significant if P ＜ 0.05.Results In comparison with the other two groups,levels of plasma LPS and D-lactate in SAP group increased markedly at 24 h after operation,which indicated that the penetrability of intestinal mucosal barrier increased (P ＜ 0.05 ). The expression of HMGB1 in the intestinal mucosa of SAP group increased significantly compared with control group (P ＜ 0.05).Whereas,the expression of occludin was significantly lower than control group (P ＜0.05).Compared with SAP group,the expression of HMGB1 was lower and the expression of occludin was higher in PDTC group ( P ＜ 0.05).Conclusions The over-expression of HMGB1 could down regulate the expression of occludin in intestinal tissues of SAP rats,and thus mediate an increase in penetrability of intestinal mucosal barrier.As PDTC inhibited the expression of HMGB1,the expression of occludin protein was up-regulated and the function of intestinal mucosal barrier was improved.
