清胰汤对L-精氨酸诱发的重症急性胰腺炎小鼠胰腺p-STAT3表达的影响

Effect of Qingyi decoction on p-STAT3 expression in pancreas of mice with severe acute pancreatitis induced by L-arginine

目的:观察L-精氨酸(L-Arg)诱发的重症急性胰腺炎小鼠胰腺组织p-STAT3表达的变化,以及清胰汤对p-STAT3表达的影响,从而探讨STAT3在急性胰腺炎中的作用和清胰汤治疗急性胰腺炎的机制。方法:健康雄性成年昆明种小鼠30只,随机分为3组(n=10):对照组、模型组和清胰汤组。除对照组外,其余各组给予腹腔注射20%L-精氨酸(3 g/kg,间隔1 h再注射1次);清胰汤组在第2次腹腔注射20%L-Arg 30 min后给予清胰汤浓缩液灌胃(10 mL/kg),之后每天灌胃2次。在造模后72 h麻醉处死动物检测血清淀粉酶活性;取胰腺组织计算胰腺湿重比,HE染色观察胰腺病理学改变;取肺组织匀浆检测髓过氧化物酶(MPO)的活性,HE染色观察肺病理学改变;Western blotting及real-time PCR分别检测胰腺组织p-STAT3蛋白及单核细胞趋化蛋白-1(MCP-1)mRNA的表达变化。结果:L-Arg诱发急性胰腺炎72 h后,血清淀粉酶活性明显升高、胰腺湿重比增加、肺组织MPO显著增加,与对照组比较差异显著(P<0.05);而清胰汤组血清淀粉酶的活性、胰腺湿重比、MPO水平明显降低,与模型组相比差异显著(P<0.01);模型组72 h胰腺及肺可见明显病理损伤,胰腺组织p-STAT3蛋白及MCP-1 mRNA的表达明显增强;清胰汤治疗组胰腺及肺病理损伤减轻,胰腺组织p-STAT3蛋白及MCP-1 mRNA的表达减少。结论:L-Arg诱发的重症急性胰腺炎小鼠胰腺组织STAT3蛋白表达明显增加,STAT3活化可能参与了L-Arg诱发的急性胰腺炎进展;抑制胰腺STAT3活化是清胰汤治疗急性胰腺炎的作用机制之一。

AIM： To explore the role of STAT3 signaling pathway in acute pancreatitis induced by L-arginine,and the mechanisms of Qingyi decoction（QYD） treatment on severe acute pancreatitis（SAP）.METHODS： The Kunming mice were randomly divided into 3 groups（n=10）： control group,SAP group and QYD treatment group（SAP＋QYD）.The mice in SAP group and SAP＋QYD group were intraperitoneally injected with 20% L-arginine（3 g/kg,bid）.The mice in SAP＋QYD group were also administered intragastrically with QYD（10 mL/kg） 30 min after the second injection of 20% L-arginine and twice a day for the following 2 days.The mice were anesthetized and sacrificed 72 h after SAP induction.The activity of amylase was measured in serum,the relative pancreatic weight was assayed,and the activity of myeloperoxidase（MPO） was analyzed to evaluate the neutrophil infiltration in lung tissues.The morphology of pancreas and lung was observed.The protein of pancreas was extracted to detect the expression of p-STAT3 by Western blotting.The mRNA expression of monocyte chemoattractant protein-1（MCP-1） was determined by real-time PCR.RESULTS： Compared with control group at 72 h,L-arginine induced SAP with increased serum amylase activity,pancreatic wet weight ratio and MPO activity in lung tissues（P0.05）.In SAP＋QYD group,the activity of amylase,pancreatic wet weight ratio and MPO levels were significantly lower than those in SAP group（P0.01）.Compared with control group at 72 h,the pancreas and lung were obvious injured,the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas tissues increased significantly in SAP group.Compared with SAP group,the pathologic damage of the pancreas and lung tissues,the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas were significantly reduced in SAP＋QYD group.CONCLUSION： The expression of p-STAT3 in pancreas increases in the mice with SAP induced by L-arginine.The activation of STAT3 may take part in the development of SAP.Inhibition of STAT3 activation in pancreas is one of the mechanisms of QYD treatment for SAP.