摘要
目的探讨Janus激酶/信号转导和转录激活因子3(JAK/STAT3)信号转导通路在重症急性胰腺炎(SAP)肺泡Ⅱ型上皮细胞损伤中的作用。方法用牛磺胆酸钠建立SAP大鼠模型,取血清备用。将经原代培养的肺泡Ⅱ型上皮细胞随机分组,对照组加入不含SAP大鼠血清的Dulbecco改良Eagle培养基(DMEM)培养液;SAP组加入含SAP大鼠血清的DMEM培养液;SAP+AG490组用JAK激酶抑制剂AG490预处理细胞,加入含SAP大鼠血清的DMEM培养液。用电泳迁移率改变分析法(EMSA)检测STAT3活化状态;逆转录-聚合酶链反应(RT—PCR)检测mRNA表达;蛋白质免疫印迹法(Westernblotting)检测蛋白水平;流式细胞技术检测肺泡表面活性物质相关蛋白C(SP—C)表达和肺泡Ⅱ型上皮细胞凋亡。结果与对照组比较,SAP组STAT3活性增强,STAT3 mRNA和蛋白表达均增强,SP—C蛋白表达下降(2hSP—C荧光指数0.69±0.02比1.02±0.03,P〈0.01),肺泡Ⅱ型上皮细胞凋亡增加[(11.55±1.10)%比(5.30±0.36)%,P〈0.053;与SAP组比较,SAP+AG490组STAT3活性减弱,STAT3mRNA和蛋白表达减弱,SP—C蛋白表达下降(2hSP—C荧光指数0.48±0.10比0.69±0.02,P〈0.01),肺泡Ⅱ型上皮细胞凋亡增加[(13.92±0.82)%比(11.55±1.10)%,P〈O.05]。结论提示JAK/STAT3信号转导通路参与了SAP肺泡Ⅱ型上皮细胞损伤的病理生理过程。
Objective To study the role of Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) in injury of alveolar type Ⅱepithelial cells (AT Ⅱ ) in severe acute pancreatitis (SAP). Methods The rat model of SAP was reproduced intraductal injection of sodium tauroeholate. Serum was collected for examinations. AT I cells in primary culture were randomized to be treated with serum from rats with SAP (SAP group), or SAP serum+AG490 (JAK inhibitor, SAP-bAG490 group), while the normal cell control was cultured with Dulbecco improvemed Eagle medium (DMEM), AT Ⅱ cells were collected after the treatment to determine the activation of STAT3 by electrophoretic mobility shift assay (EMSA), STAT3 mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR), STAT3 protein expression by Western blotting, surfactant protein C (SP-C) level in AT Ⅱ and apoptosis of AT Ⅱ with flow cytometry. Results Compared with control group, activation of STAT3 was enhanced in SAP group, and expression of STAT3 mRNA and protein was also enhanced, level of SP-C protein declined (2 hours SP-C fluorescence index 0.69± 0.02 vs. 1.02 ±0.03, P 〈 0.01), and apoptosis of AT Ⅱ increased [(11.55 ± 1.10)% vs. (5.30±0. 36)%, P〈0.05]. Compared with SAP group, activation of STAT3 was attenuated in SAP+AG490 group, expression of STAT3 mRNA and protein was lowered, and level of SP-C protein declined (2 hours SP-C fluorescence index 0.48± 0. 10 vs. 0. 69 ±0.02, P〉0.01), and the apoptosis of AT Ⅱincreased [(13.92±0.82)% vs. (11.55±1.10)%, P〈0.05]. Conclusion The results suggest that JAK/STAT3 pathway might be involved in the pathogenesis of injury to AT Ⅱ in SAP.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2009年第2期107-110,共4页
Chinese Critical Care Medicine