腺苷酸活化蛋白激酶抑制剂对急性水肿性胰腺炎大鼠胰腺外分泌功能及肠道通透性的影响

Effect of AMP-activated protein kinase inhibitor on pancreatic exocrine function and intestinal permeability in rats with acute edematous pancreatitis

目的研究腺苷酸活化蛋白激酶(AMPK)抑制剂(Compound C)对急性水肿性胰腺炎(AEP)大鼠胰腺外分泌功能及肠道通透性的影响。方法将SD大鼠随机分为模型组、实验组和对照组,每组15只,均于腹股沟皮下注射雨蛙素建立AEP大鼠模型,实验组灌胃10μmol·L^-1 Compound C 2 mL,对照组灌胃0.16 g·mL^-1L-谷氨酰胺2 mL,模型组灌胃等量生理盐水,连续干预2 d。用全自动生化分析仪检测胆胰液中分泌物浓度,以荧光法检测大鼠肠黏膜通透性,以蛋白质印迹法检测大鼠肠黏膜闭合蛋白(occludin)和AMPK蛋白表达。结果实验组和对照组大鼠胆胰液中Ca^2+浓度较模型组升高,淀粉酶(AMS)和脂肪酶(LIP)浓度较模型组降低,差异均有统计学意义(均P<0.05)。实验组和对照组大鼠肠黏膜通透性较模型组明显降低,差异有统计学意义(P<0.01)。模型组、实验组和对照组肠黏膜occludin蛋白相对表达量分别为0.59±0.16,1.23±0.22,0.96±0.15,AMPK蛋白相对表达量分别为1.55±0.17,0.66±0.18,0.99±0.21。实验组和对照组与模型组比较,occludin蛋白表达量升高,AMPK蛋白表达量降低(P<0.01)。结论AMPK抑制剂可改善AEP大鼠胰腺外分泌功能及肠黏膜通透性,AMPK通路活化是通过负性调控occludin蛋白的表达促进肠黏膜通透性的增强。

Objective To investigate the effect of AMP-activated protein kinase(AMPK)inhibitor(Compound C)on pancreatic exocrine function and intestinal permeability in rats with acute edematous pancreatitis(AEP).Methods SD rats were randomly divided into model group,test group and control group,15 rats in each group.AEP rats model was established by rainforest prime subcutaneous injections into the groin with caerulein,test group was gavage with 10μmol·L^-1Compound C 2 mL,control group was gavage with 0.16 g·mL^-1 L-glutamine 2 mL,model group was gavage with the same amount of normal saline.All were intervened for 2 d.Automatic analyzer was used to detect secretions of bile and pancreatic juice,intestinal mucosal permeability of rat was detected by fluorescence method,occludin and AMPK protein expression were detected by Western blotting.Results The concentrations of Ca^2+in test group and control group were higher than that in model group,while amylase(AMS)and lipase(LIP)were lower than those in model group,all with significant difference(all P<0.05).Intestinal mucosal permeability of test group and control group were significantly lower than model group(P<0.01).The relative expression of occludin protein in model group,test group and control group were 0.59±0.16,1.23±0.22,0.96±0.15,relative expression of AMPK protein were 1.55±0.17,0.66±0.18,0.99±0.21.Compared with model group,the expression of occludin protein and AMPK protein in test group and control group were decreased with significant(P<0.01).Conclusion AMPK inhibitor can improve pancreatic exocrine function and intestinal mucosal permeability in AEP rats,the mechanism is AMPK pathway activation in AEP rats,intestinal mucosal permeability enhancement can be enhanced by negative regulation of occludin protein expression.