氧化应激及凋亡与重症急性胰腺炎肠屏障功能障碍
양화응격급조망여중증급성이선염장병장공능장애
Oxidative stress and apoptosis in gut barrier dysfunction of severe acute pancreatitis
  • 万方数据
    中华急诊医学杂志 중화급진의학잡지
    CHINESE JOURNAL OF EMERGENCY MEDICINE
    2012年 10期 1088-1092 ,共5页

    田锐%许飞%王瑞兰%谢晖%孟潇潇%钱永兵%金卫%胡家昌%周志刚%俞康龙

    전예%허비%왕서란%사휘%맹소소%전영병%금위%호가창%주지강%유강룡

    全身炎症反应综合征%炎症因子%缺血-再灌注损伤%氧自由基%氧化应激%凋亡%重症急性胰腺炎%肠道屏障功能障碍 全身炎癥反應綜閤徵%炎癥因子%缺血-再灌註損傷%氧自由基%氧化應激%凋亡%重癥急性胰腺炎%腸道屏障功能障礙
    전신염증반응종합정%염증인자%결혈-재관주손상%양자유기%양화응격%조망%중증급성이선염%장도병장공능장애
    Systemic inflammatory response syndrome (SIRS)%Inflammatory factors%Ischemia-reperfusion injury%Oxygen radicals%Oxidative stress%Apoptosis%Severe acute pancreatitis%Gut barrier dysfunction
    目的 通过动物实验,观察重症急性胰腺炎(severe acute pancreatitis,SAP)肠道屏障功能变化,探讨炎症因子释放、肠黏膜氧化应激及凋亡在肠屏障功能障碍中的作用.方法 上海交通大学附属第一人民医院动物实验中心内,24只BALB/c小鼠,随机数字法分为2组,SAP组:以雨蛙素联合脂多糖腹腔注射法诱导,先腹腔内注射雨蛙素50 μg/kg,连续6次,每次间隔1h,在末次雨蛙素注射同时,腹腔内注射脂多糖10 mg/kg(LPS E.Coli)；对照(假手术)组:每小时一次腹腔注射生理盐水2 ml,共6次.两组动物分2批(每批6只/组)分别于建模后4h及8h,麻醉后打开腹腔取血及标本.观察小鼠胰腺及肠道病理变化并予评分,测定小鼠血二胺氧化酶(diamine oxidase,DAO)、淀粉酶、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量,测定肠黏膜丙二醛( malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)、还原型谷胱甘肽( glutathione,GSH)含量及黄嘌呤氧化酶(xanthine oxidase,XO)活力,检测小鼠肠黏膜细胞caspase-3酶活性,脱氧核糖核苷酸末端转移酶介导的缺口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)法检测小鼠肠黏膜凋亡细胞并计算凋亡指数.以PASW 18.0软件对数据进行方差分析及t检验,明确上述检测指标在两组小鼠间差异是否有统计学意义,从而明确重症急性胰腺炎肠道屏障功能障碍的机制.结果 建模后4h及8h,SAP组小鼠胰腺出血、坏死、炎症细胞浸润较对照组严重,胰腺病理评分与对照组比较差异具有统计学意义(P＜0.01),血淀粉酶含量与对照组比较差异具有统计学意义(P＜0.05)；肠组织病理评分及血DAO质量浓度与对照组比较差异具有统计学意义(P＜0.01)；血TNF-α质量浓度与对照组比较差异具有统计学意义(P＜0.01)；肠黏膜MDA含量及XO活力与对照组比较差异具有统计学意义(P＜0.01),肠黏膜SOD含量与对照组比较差异具有统计学意义(P＜0.01)、GSH含量与对照组比较差异具有统计学意义(P＜0.05)；肠黏膜细胞caspase-3酶活性及凋亡指数与对照组比较差异具有统计学意义(P＜0.01).结论 重症急性胰腺炎发病后,TNF-α等炎症因子瀑布样释放,导致肠黏膜缺血-再灌注损伤,形成严重的氧化应激反应,进一步激活caspase-3通路,导致肠黏膜细胞凋亡增加,是肠黏膜屏障功能受损的重要机制.
    목적 통과동물실험,관찰중증급성이선염(severe acute pancreatitis,SAP)장도병장공능변화,탐토염증인자석방、장점막양화응격급조망재장병장공능장애중적작용.방법 상해교통대학부속제일인민의원동물실험중심내,24지BALB/c소서,수궤수자법분위2조,SAP조:이우와소연합지다당복강주사법유도,선복강내주사우와소50 μg/kg,련속6차,매차간격1h,재말차우와소주사동시,복강내주사지다당10 mg/kg(LPS E.Coli)；대조(가수술)조:매소시일차복강주사생리염수2 ml,공6차.량조동물분2비(매비6지/조)분별우건모후4h급8h,마취후타개복강취혈급표본.관찰소서이선급장도병리변화병여평분,측정소서혈이알양화매(diamine oxidase,DAO)、정분매、종류배사인자-α(tumor necrosis factor-α,TNF-α)함량,측정장점막병이철( malondialdehyde,MDA)、초양화물기화매(superoxide dismutase,SOD)、환원형곡광감태( glutathione,GSH)함량급황표령양화매(xanthine oxidase,XO)활력,검측소서장점막세포caspase-3매활성,탈양핵당핵감산말단전이매개도적결구말단표기(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)법검측소서장점막조망세포병계산조망지수.이PASW 18.0연건대수거진행방차분석급t검험,명학상술검측지표재량조소서간차이시부유통계학의의,종이명학중증급성이선염장도병장공능장애적궤제.결과 건모후4h급8h,SAP조소서이선출혈、배사、염증세포침윤교대조조엄중,이선병리평분여대조조비교차이구유통계학의의(P＜0.01),혈정분매함량여대조조비교차이구유통계학의의(P＜0.05)；장조직병리평분급혈DAO질량농도여대조조비교차이구유통계학의의(P＜0.01)；혈TNF-α질량농도여대조조비교차이구유통계학의의(P＜0.01)；장점막MDA함량급XO활력여대조조비교차이구유통계학의의(P＜0.01),장점막SOD함량여대조조비교차이구유통계학의의(P＜0.01)、GSH함량여대조조비교차이구유통계학의의(P＜0.05)；장점막세포caspase-3매활성급조망지수여대조조비교차이구유통계학의의(P＜0.01).결론 중증급성이선염발병후,TNF-α등염증인자폭포양석방,도치장점막결혈-재관주손상,형성엄중적양화응격반응,진일보격활caspase-3통로,도치장점막세포조망증가,시장점막병장공능수손적중요궤제.
    Objective By means of animal study,investigated the gut barrier function in severe acute pancreatitis ( SAP),and role of inflammatory factors releasing,gut mucosa oxidative stress,cell apoptosis in it.Methods The animal experiment was done in the animal center of first people' s hospital,shanghai jiaotong university.Twenty four BALB/c mice were randomized ( random number) divided into two groups with twelve mice each group.The SAP group,mice received six intraperitoneal injections of cerulein at 1-hour intervals, the dose was 50μg/kg, then given one intraperitoneal injection of 10 mg/kg lipopolysaccharide ( LPS from E.Coli) for the induction of severe acute pancreatitis.The control ( sham operation) group,the mice received intraperitoneal injection of 2 ml normal saline for six times at 1-hour intervals.All the animals of each group were averaged to two batches,4 h and 8h after being operated respectively,to be anesthetized and adopted blood and tissue specimen.Then we observed the pathological change of pancreas and gut,scored it.We measured the blood value of diamine oxidase ( DAO),amylase and tumor necrosis factor-α (TNF-α).We detected content of malondialdehyde (MDA),superoxide dismutase (SOD),glutathione (GSH) and activity of xanthine oxidase (XO) in gut mucosa.We detected the casepase-3 activity and cell apopotosis by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in gut mucosa,and conculated the apopotosis index (AI).Then using the PASW 18.0 software,we analyzed the data by anova and t-test,to make sure if the values were statistically different between the two groups and the mechanism of gut barrier dysfunction in panreatitis.Results At 4 h and 8 h after operation,the SAP-group-mice had significantly higher pancreas pathological score (P ＜0.01 ),blood amylase value ( P ＜ 0.05 ),gut pathological score and blood DAO and TNF-α value ( P ＜0.01 ),compared with the contral-group-mice.The gut mucosa MDA content and XO activity of mice in SAP group were significantly higher than which in control group ( P ＜ 0.01 ). The SAP-group-mice had significantly lower gut mucosa SOD content ( P ＜ 0.01 ) and GSH content ( P ＜ 0.05 ),compared with the contral-group-mice.The gut mucosa cells of mice in SAP group had significantly higher caspase-3 activity and apoptosis index than which in control group ( P ＜ 0.01 ).Conclusions In severe acute pancreatitis,inflammatory factors such as TNF-αwere waterfall-style released,induced gut mucosa suffer from ischemia-reperfusion injury,then serious oxidative stress developed in mucosa and activated caspase-3 pathway,inducing gut mucosa cells apoptose seriously,which was an important mechanism of gut barrier dysfunction.
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