调控高迁移组蛋白1诱导大鼠肺泡巨噬细胞表达诱生型一氧化氮合酶的信号通路  

Signaling Pathways in Expression of Inducible Nitric Oxide Synthase Induced by High Mobility Group Box 1 in Rat Alveolar Macrophages

作　　者：余跃[1] 任大宾[1] 孙仁宇[1] 王士雯[2] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所病理生理学系 [2]Institute of Geriatric Cardiology of Chinese PLA General Hospital,Beijing100853

出　　处：《中国医学科学院学报》2006年第6期781-785,共5页Acta Academiae Medicinae Sinicae

基　　金：国家重点基础研究发展计划项目(973计划)(G2000057004)~~

摘　　要：目的观察细胞外信号调节激酶(ERK)、p38分裂原激活的蛋白激酶(p38MAPK)及核因子-κB(NF-κB)在高迁移组蛋白1(HMGB1)诱导大鼠肺泡巨噬细胞表达诱生型一氧化氮合酶(iNOS)中的作用。方法体外培养的大鼠原代肺泡巨噬细胞(PRAMs)分别与ERK抑制剂PD98059、p38MAPK抑制剂SB203580和NF-κB抑制剂PDTC,以及PD98059联合SB203580共孵育2h后,培养基中分别加入重组人HMGB1作用6h。采用逆转录聚合酶链式反应检测培养细胞中iNOSmRNA表达,用Greiss法测定培养上清中NO2-/NO3-的含量。结果PD98059、SB203580和PDTC均可显著下调HMGB1诱导PRAMs表达iNOSmRNA和产生NO(P<0·05)。联合使用PD98059和SB203580可增强抑制iNOSmRNA表达和NO产生(P<0·05)。结论信号分子ERK、p38MAPK和NF-κB均参与了HMGB1诱导大鼠肺泡巨噬细胞表达iNOS和产生NO。Objective To explore roles of extracellular signal-regulated kinase （ERK） 1/2, p38 mitogen activated protein kinase （ p38 MAPK） and nuclear factor （NF） -κB in expression of inducible nitric oxide synthase （iNOS） in rat alveolar macrophages induced by high mobility group box 1 （ HMGB1 ）. Methods Primary rat alveolar macrophages （PRAMs） cultured in vitro were incubated with PD98059 （ inhibitor against ERK）, SB203580 （ inhibitor against p38 MAPK）, PDTC （ inhibitor against NF-κB ）, or PD98059 plus SB203580 for 2 hours, respectively. HMGB1 was added into the cultures and incubated with cells for 6 hours. Total RNA of PRAMs was extracted and iNOS mRNA expression was semi-quantified with reverse transcription-polymerase chain reaction （RT-PCR）. Greiss reaction was applied to determine nitrite/nitrate （ NO2^-/NO3^- ） concentration in PRAMs culture supernatants. Results Expression of iNOS mRNA and NO production in PRAMs culture supematants were down-regulated by inhibition of ERK or p38 MAPK by PD98059 or SB203580, respectively （P 〈 0. 05 ）. Moreover, inhibition of iNOS expression and NO production was observed after simultaneous pretreatment with PD98059 and SB203580 （ P 〈 0. 05 ）. Expression of iNOS mRNA in PRAMs and NO production in PRAMs culture supernatants were down-regulated by inhibition of NF-κB by PDTC （P 〈 0. 05）. Conclusion Cellular signal molecules of ERK, p38 MAPK, and NF-κB all participate in the expression of iNOS and NO production in PRAMs induced by HMGB1.

关 键 词：高迁移组蛋白1 肺泡巨噬细胞 诱生型一氧化氮合酶 

分 类 号：R363[医药卫生—病理学]