细胞外调节蛋白激酶在大鼠急性坏死性胰腺炎发病机制中的作用  

Role of extracellular regulated kinase signal transduction pathway in the pathogenesis of acute necrotizing pancreatitis

作　　者：施新岗[1] 许永春[1] 满晓华[1] 金晶[1] 龚燕芳[1] 屠振兴[1] 李兆申[1] 

机构地区：[1]第二军医大学长海医院消化内科,上海200433

出　　处：《中华胰腺病杂志》2010年第2期116-119,共4页Chinese Journal of Pancreatology

摘　　要：目的 研究细胞外调节蛋白激酶（extracellular regulated kinase 1/2,ERK1/2）信号转导通路在大鼠急性坏死性胰腺炎（ANP）时的变化规律,探讨ERK1/2特异性抑制剂PD98059对ANP时胰腺的保护作用.方法 以5%牛磺胆酸钠胰胆管逆行注射建立ANP模型,5只大鼠作为正常组,余75只大鼠按数字表法随机分为假手术组、ANP组及PD98059组,各25只.术后15 min、30 min、1 h、3 h和6 h分批处死动物,取血及胰腺组织.常规胰腺病理检查并评分;ELISA方法检测大鼠血清IL-1β、TNF-α水平;酶化学法检测胰腺组织内MPO活性;Western blotting法检测胰腺组织磷酸化ERK1/2的表达量.结果 假手术组胰腺无明显病理改变;ANP组胰腺损伤明显,3 h的病理评分为9.9±0.4,PD98059组胰腺损伤减轻,3 h病理评分为4.0±0.4,与ANP组相差显著（P〈0.05）.假手术组、ANP组和PD98059组大鼠3 h时血清TNF-α水平分别为（65.8±20.5）pg/ml、（286.5±50.3）pg/ml、（180.4±32.9）pg/ml;IL-1β水平为（85.8±25.5）pg/ml、（293.8±46.3）pg/ml、（200.5±33.6）pg/ml;胰腺MPO活性为（0.19±0.02）U/g、（0.61±0.05）U/g、（0.52±0.03）U/g.ANP组和PD98059组均显著高于假手术组,而PD98059组又显著低于ANP组（P值均〈0.05）.正常大鼠胰腺磷酸化ERK1/2表达量为1100±141;ANP组15 min、30 min时磷酸化ERK1/2表达量分别为5300±486、5621±384,1 h开始下降,6 h时几乎与假手术组相似;PD98059组造模后15 min、30 min的磷酸化ERK1/2表达量分别为4200±370、3600±290,显著低于同时间点ANP组（P值均〈0.01）.结论 ERK1/2信号转导通路参与大鼠ANP发病机制,PD98059干预可减少IL-1β、TNF-α的产生,降低胰腺组织MPO活性,改善胰腺的病理损伤程度.Objective To investigate the changes of extraceUular regulated kinase 1/2 （ERK1/2） phosphorylation and assess the effects of blocking the ERK1/2 phosphorylation on rats with acute necrotizing pancreatitis （ANP）. Methods The ANP model was induced by retrograde injection of 5% sodium tanrocholate into the biliary and pancreatic duct. 5 rats were treated as normal control. Other 75 Sprague-Dawley （SD） rots were randomly divided into sham operations（SO） group （n =25）, ANP group （n =25） and PD98059 group （n =25）. The rats were sacrificed at 15 min, 30 min, 1 h, 3 h and 6 h after ANP induction, the blood and pancreatic sample were taken. Pathological changes of pancreas were observed with light microscope and scored. The serum level of TNF-α and IL-1β was determined by ELISA. MPO activities in pancreas were measured by enzyme chemistry assay. Western blotting was performed to determine the phosphorylations of ERK1/2 in the pancreas homogenates. Results There was no significant pathologic changes in rats of SO group;but significant injuries occurred in ANP group, the pathologic score at 3 h was 9.9 ± 0.4;the extent of injuries attenuated in PD98059 group, the pathologic score at 3 h was 4.0 ± 0.4 （P 〈 0.05）. The serum levels of TNF-α at 3 h in SO, ANP and PD98059 groups were （65.8 ± 20.5） pg/ml, （286.5 ± 50.3） pg/ml, （180.4±32.9）pg/ml, respectively;the serum levels of IL-1β at 3 h in SO, ANP and PD98059groups were （85.8 ± 25.5） pg/ml, （293.8 ± 46.3） pg/ml, （200. 5 ± 33.6） pg/ml, respectively;MPOactivities in pancreas were （0. 19 ± 0.02）U/g, （0.61±0.05）U/g, （0.52±0.03） U/g, and the values in ANP and PD98059 groups were significantly higher than those in SO group, while the values in PD98059 group were significantly lower than those in ANP group （P 〈 0.01）. The expression of ERK1/2 phosphorylation in normal pancreas was 1100 ± 141, the expressions of ERK1/2 phosphorylation in ANP group at 15 min, 30 min were 5300 ± 486, 5621 ± 38

关 键 词：胰腺炎 急性胰腺炎 细胞外信号调节激酶 胰腺损伤 

分 类 号：R576[医药卫生—消化系统]