中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2009年
2期
107-110
,共4页
钟敦璟%赵晓晏%郝嘉%郭红
鐘敦璟%趙曉晏%郝嘉%郭紅
종돈경%조효안%학가%곽홍
Janus激酶/信号转导和转录激活因子3%胰腺炎,急性%肺泡Ⅱ型上皮细胞%肺泡表面活性物质蛋白-C%细胞凋亡
Janus激酶/信號轉導和轉錄激活因子3%胰腺炎,急性%肺泡Ⅱ型上皮細胞%肺泡錶麵活性物質蛋白-C%細胞凋亡
Janus격매/신호전도화전록격활인자3%이선염,급성%폐포Ⅱ형상피세포%폐포표면활성물질단백-C%세포조망
Janus kinase/signal transducer and activator of transcription 3%acute pancreatitis%alveolar type I epithelial cell%surfactant protein C%apoptosis
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;逆转录一聚合酶链反应(RT-PCR)检测mRNA表达;蛋白质免疫印迹法(Western blotting)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(2 h SP-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,STAT3 mRNA和蛋白表达减弱,SP-C蛋白表达下降(2 h SP-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＜0.053.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.
目的 探討Janus激酶/信號轉導和轉錄激活因子3(JAK/STAT3)信號轉導通路在重癥急性胰腺炎(SAP)肺泡Ⅱ型上皮細胞損傷中的作用.方法 用牛磺膽痠鈉建立SAP大鼠模型,取血清備用.將經原代培養的肺泡Ⅱ型上皮細胞隨機分組,對照組加入不含SAP大鼠血清的Dulbecco改良Eagle培養基(DMEM)培養液;SAP組加入含SAP大鼠血清的DMEM培養液;SAP+AG490組用JAK激酶抑製劑AG490預處理細胞,加入含SAP大鼠血清的DMEM培養液.用電泳遷移率改變分析法(EMSA)檢測STAT3活化狀態;逆轉錄一聚閤酶鏈反應(RT-PCR)檢測mRNA錶達;蛋白質免疫印跡法(Western blotting)檢測蛋白水平;流式細胞技術檢測肺泡錶麵活性物質相關蛋白C(SP-C)錶達和肺泡Ⅱ型上皮細胞凋亡.結果 與對照組比較,SAP組STAT3活性增彊,STAT3 mRNA和蛋白錶達均增彊,SP-C蛋白錶達下降(2 h SP-C熒光指數0.69±0.02比1.02±0.03,P＜0.01),肺泡Ⅱ型上皮細胞凋亡增加[(11.55±1.10)%比(5.30±0.36)%,P＜0.053;與SAP組比較,SAP+AG490組STAT3活性減弱,STAT3 mRNA和蛋白錶達減弱,SP-C蛋白錶達下降(2 h SP-C熒光指數0.48±0.10比0.69±0.02,P＜0.01),肺泡Ⅱ型上皮細胞凋亡增加[(13.92±0.82)%比(11.55±1.10)%,P＜0.053.結論 提示JAK/STAT3信號轉導通路參與瞭SAP肺泡Ⅱ型上皮細胞損傷的病理生理過程.
목적 탐토Janus격매/신호전도화전록격활인자3(JAK/STAT3)신호전도통로재중증급성이선염(SAP)폐포Ⅱ형상피세포손상중적작용.방법 용우광담산납건립SAP대서모형,취혈청비용.장경원대배양적폐포Ⅱ형상피세포수궤분조,대조조가입불함SAP대서혈청적Dulbecco개량Eagle배양기(DMEM)배양액;SAP조가입함SAP대서혈청적DMEM배양액;SAP+AG490조용JAK격매억제제AG490예처리세포,가입함SAP대서혈청적DMEM배양액.용전영천이솔개변분석법(EMSA)검측STAT3활화상태;역전록일취합매련반응(RT-PCR)검측mRNA표체;단백질면역인적법(Western blotting)검측단백수평;류식세포기술검측폐포표면활성물질상관단백C(SP-C)표체화폐포Ⅱ형상피세포조망.결과 여대조조비교,SAP조STAT3활성증강,STAT3 mRNA화단백표체균증강,SP-C단백표체하강(2 h SP-C형광지수0.69±0.02비1.02±0.03,P＜0.01),폐포Ⅱ형상피세포조망증가[(11.55±1.10)%비(5.30±0.36)%,P＜0.053;여SAP조비교,SAP+AG490조STAT3활성감약,STAT3 mRNA화단백표체감약,SP-C단백표체하강(2 h SP-C형광지수0.48±0.10비0.69±0.02,P＜0.01),폐포Ⅱ형상피세포조망증가[(13.92±0.82)%비(11.55±1.10)%,P＜0.053.결론 제시JAK/STAT3신호전도통로삼여료SAP폐포Ⅱ형상피세포손상적병리생리과정.
Objective To study the role of Janus kinase/signal transducer and activator of transcription 3(JAK/STAT3)in injury of alveolar typeⅡepithelial cells(ATⅡ)in severe acute pancreatitis(SAP).Methods The rat model of SAP was reproduced intraductal injection of sodium taurocholate.Serum was collected for examinations.ATⅡcells in primary culture were randomized to be treated with serum from rats with SAP(SAP group),or SAP serum+AG490(JAK inhibitor,SAP+AG490 group),while the normal cell control was cultured with Dulbecco improvemed Eagle medium(DMEM).ATⅡcells 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 ATⅡand apoptosis of ATⅡwith 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 ATⅡincreased[(11.55±1.10)%vs.(5.30±0.36)%,P＜0.053.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 Ⅱ increased[(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 ATⅡin SAP.
