磷脂酰肌醇3激酶/蛋白激酶B信号转导通路在重症急性胰腺炎中性粒细胞活化中的作用  

Role of phosphatidylinositol 3-kinase/protein kinase B signal transduction pathway in activation of neutrophils during severe acute pancreatitis

作　　者：张成[1] 栾正刚[2] 葛春林[3] 郭仁宣[3] 

机构地区：[1]中国医科大学附属第一医院普外二科辽宁省沈阳市,110001 [2]中国医科大学附属第一医院ICU室,辽宁省沈阳市110001 [3]中国医科大学附属第一医院普外二科,辽宁省沈阳市110001

出　　处：《世界华人消化杂志》2006年第20期1987-1991,共5页World Chinese Journal of Digestology

摘　　要：目的：探讨磷脂酰肌醇3激酶/蛋白激酶B (PI3K/PKB)信号转导通路在重症急性胰腺炎中性粒细胞活化中的作用.健康成年♂SD大鼠30只,随机分为对照组、SAP组和SAP+wortmannin组,每组10只.胆胰管内逆行注射法制作SAP模型,模型制成6h后采用密度梯度离心方法分离出中性粒细胞,用ELISA法检测血中TNF-α和IL- 1β的含量,RT-PCR技术检测中性粒细胞内TNF-α和IL-1βmRNA的变化,Western blot法检测细胞中PKB的活性变化.结果：胰腺炎大鼠血中TNF-α和IL-1β的含量较对照组明显升高(150.8±31.7ng/L vs 22.4±5.2ng/L,P＜0.01；217.5±38.4ng/L vs 43.7±9.6ng/L,P＜0.01),同时中性粒细胞内TNF-α和IL-1βmRNA及p-PKB的水平也明显升高(0.72±0.06 vs 0.19±0.03,P＜0.01；0.84±0.11 vs 0.14±0.04,P＜0.01；0.63±0.08 vs 0.34±0.03,P＜0.01),wortmannin能显著抑制血中TNF-α(70.9±11.3 vs 150.8±31.7ng/L)和IL-1β(93.0±11.5 vs 217.5±38.4ng/L)含量,中性粒细胞中TNF-α和IL-1βmRNA(0.32±0.04 vs 0.72±0.06及0.22±0.04 vs 0.84±0.11,P<0.01)的表达及PKB活性(0.38±0.06 vs 0.63±0.08,P＜0.01)也显著下降.结论：PI3K/PKB信号转导通路介导了重症急性胰腺炎大鼠中性粒细胞的活化及促炎性细胞因子TNF-α和IL-1β的产生.AIM： To investigate the role of phosphatidylinositol 3-kinase/protein kinase B （PI3K/PKB） signal transduction pathway in the activation of neutrophils during severe acute pancreatitis （SAP）. METHODS： Thirty male health adult Sprague Dawley rats were averagely randomized into group A, B and C. SAP model was established in group B and C by retrograde injection of 50 g/L sodium taurocholate into the pancreatic and biliary duct. The rats in group C were treated with wortmannin 4 h before modeling, and those in group A served as the controls. All the rats were killed 6 h after modeling, and the neutrophils were isolated by density gradient centrifugation. The serum levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were analyzed by enzyme-linked immunosorbent assay （ELISA）. The expression of TNF-α and IL-1β mRNA were detected by RT-PCR and the activity of phosphorylated PKB （p-PKB） was examined by Western blot. RESULTS： The serum levels of TNF-α and IL- 1β were significantly higher in group B than those in group A （150.8 ± 31.7 ng/L vs 22.4 ± 5.2 ng/L, P 〈 0.01; 217.5 ± 38.4 ng/L vs 43.7 ± 9.6 ng/L, P 〈 0.01）, and mRNA levels of TNF-α and IL-1β were also markedly increased （0.72 ± 0.06 vs 0.19 ± 0.03, P 〈 0.01; 0.84 ± 0.11 vs 0.14 ± 0.04, P 〈 0.01）. However, the serum and mRNA levels of TNF-α and IL-1β were all significantly decreased in group C in comparison with those in group B （TNF-α： 70.9 ± 11.3 ng/L, 0.32 ± 0.04; IL-1β： 93.0 ± 11.5 ng/L, 0.22 ± 0.04; all P 〈 0.01）. Western blot showed that the activity of p-PKB was increased in group B as compared with that in group A （0.63 ± 0.08 vs 0.34 ± 0.03, P 〈 0.01）, but it was markedly down-regulated in group belong to （0.38 ± 0.06, P 〈 0.01）. CONCLUSION： PI3K/PKB signal transduction pathway is activated in the neutrophils during severe acute pancreatitis, which mediates the production of pro-inflammatory cytokines TNF-α and IL-1β.

关 键 词：磷脂酰肌醇3激酶 重症急性胰腺炎 中性 粒细胞 信号转导 

分 类 号：R657.51[医药卫生—外科学]