抑制Janus激酶／信号转导和转录激活子通路对胰腺腺泡细胞核因子-kB的影响  

The effect of inhibiting janus kinase/signal transducer and activator of transcription pathway on NF-KB expression in pancreatic acinar cell line AR42J

作　　者：陈平[1] 姚玮艳[1] 章永平[1] 乔敏敏[1] 董文杰[1] 袁耀宗[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院消化科,200025

出　　处：《中华消化杂志》2010年第3期179-183,共5页Chinese Journal of Digestion

摘　　要：目的旨在观察Janus激酶（JAK）／信号转导和转录激活子（signal transducer and activator of transcription, STAT）信号转导抑制剂对构建的急性胰腺炎体外模型中核因子（NF）-kB变化及与炎性反应的影响，为急性胰腺炎的治疗提供理论依据。方法AR42J细胞随机分为4组，对照组、雨蛙素处理组、雷帕霉素（RPM）＋雨蛙素处理组、AG490＋雨蛙素处理组，于24h收集细胞，EMSA测定NF—KB活性，免疫荧光及Western印迹法测定NF—KB亚基表达，此外，RT—PCR及Western印迹测定肿瘤坏死因子（TNF）a，白细胞介素（IL）-1β和IL-6蛋白及基因表达。结果雨蛙素处理组于24hNF-kB活性表达增加，NF—KB抑制蛋白激酶（IKK）p，NF—KBP50及NF-kBP65蛋白水平增高。免疫荧光示NF_KBP65核转位。此外TNFa、IL-1β和IL-6蛋白及mRNA表达于24h后明显升高（与对照组比较，P〈0．05）。RPM或AG490治疗组与雨蛙素处理组比较则明显降低NF—KB活性，抑制1KKβ、NF—KBP50及NF-~B表达，减少NF—KBP65核转位，进而使TNFa、IL-10和IL～6mRNA和蛋白表达下调（P〈O．05）。结论JAK／sTAT信号通路可能早期参与胰腺损伤中促炎性细胞因子释放，机制涉及NF-kB通路。关闭JAK／sTAT信号通路可作为控制急性胰腺炎中炎性反应的手段之一。Objective To investigate the impact of inhibitor of janus kinase/signal transducer and activator of transcription （JAK/STAT） on activity of NF-kB and the inflammatory response induced by cerulein in pancreatic acinar cell line AR42J. Methods AR42J cells were allocated into control group （treated with PBS）, cerulein induced group, Rapamycin combined with cerulein treated group and AG490 combined with cerulein treated group. The cells were harvested at 24 hours after treatment. The activity of NF-kB was measured by EMSA. The expression of subunits of NF-kB （P50 and P65） was detected by immunofluorescence staining and Western blotting. Furthermore, the expressions of tumor necrosis factor （TNF）a, interleukin （IL）--1β, and IL 6 genes or proteins were measured by RT-PCR and Western blotting. Results The high activity of NF- kB and increased protein levels of NF-kB inhibitor kinase （IkK）13, P50 and P65 were found in cerulein induced group. Immunofluoreseence examination showed nuclear translocation of P65. In comparison with control group, the mRNA and protein expressions of TNFα, IL-1β and IL-6 were increased 24 hours after induction （P《0.05）. Whereas Rapamycin and AG490 inhibited the activity of NF-kB, reduced the protein expressions of NF-kB P50 and P65 as well as nuclear translocation in comparison with cerulein induced group （P〈0.05）. The IkKβ in Rapamycin and AG490 treated groups were clown regulated at 24 hours after induction. Conclusions The JAK/STAT pathway involves in cerulein-induced pancreatic inflammatory response via regulating NF-kB. Blockade of JAK-STAT pathway with inhibitors may be benefit for attenuating inflammatory response.

关 键 词：胰腺炎 JANUS激酶 核转录因子B 细胞因子 AR42J细胞 

分 类 号：R576[医药卫生—消化系统]