模拟微重力对肺微血管内皮细胞窖蛋白-1和内皮型一氧化氮合酶表达的影响  

Effects of simulated microgravity on the expression of caveolin-1 protein and endothelial nitric oxide synthase in pulmonary microvascular endothelial cells

作　　者：康春燕[1] 袁明[2] 邹琳[1] 章烨[1] 李天志[1] 王俊锋[1] 刘长庭[1] 

机构地区：[1]北京解放军总医院南楼呼吸科,100853 [2]北京中国航天员科研训练中心航天医学基础与应用国家重点实验室,100094

出　　处：《解放军医学杂志》2010年第11期1322-1324,共3页Medical Journal of Chinese People's Liberation Army

基　　金：军队医药卫生“十一五”计划专项滚动课题(06Z048);武器装备预研基金(9140A1704009JB1002);民用航天科研预先研究项目(科工技[2007]302);国家高技术研究发展计划(2008AA12A220)

摘　　要：目的探讨回转模拟微重力对肺微血管内皮细胞(PMVEC)窖蛋白-1(caveolin-1)和内皮型一氧化氮合酶(eNOS)表达的影响。方法以肺微血管内皮细胞为研究对象,采用回转器模拟微重力效应。将细胞分为回转12h组、回转24h组(在30r/min条件下回转培养12、24h),以及对照12h组、对照24h组(放置于细胞回转器旁静置培养12、24h)。采用Griess法检测细胞的NO释放量,透射电镜观察细胞胞膜窖的超微结构变化,Western blotting检测eNOS和caveolin-1的表达。结果回转12h组较对照12h组NO产量和eNOS蛋白表达均有增加,回转24h组较对照24h组NO产量和eNOS蛋白表达也有增加,差异均有统计学意义(P<0.05)。由透射电镜可见,对照12h组和对照24h组PMVEC细胞膜表面均可见典型的希腊字母Ω状胞膜窖,较丰富,结构清晰;回转12h组PMVEC细胞膜上胞膜窖的结构破坏,数量减少;回转24h组PMVEC细胞膜上很难发现胞膜窖的特征性结构,数量进一步减少。Western blotting检测发现,回转12h组与对照12h组相比,回转24h组与对照24h组相比,PMVEC中caveolin-1表达均显著下降(P<0.05或P<0.01)。结论回转模拟微重力可抑制肺微血管内皮细胞中caveolin-1的表达,增强eNOS表达,提高细胞内NO的释放量。Objective To explore the effects of simulated microgravity induced by clinorotation on the expression of caveolin 1 and endothelial nitric oxide synthase （eNOS） in pulmonary microvascular endothelial ceils （PMVECs）. Methods Clinorotation with a clinostat rotator was instituted to simulate the microgravity condition, and the effects of clinorotation on the cultured PMVECs were investigated. PMVECs were cultivated under clinorotation at 30r/min for 12 and 24 hours, respectively. The control group consisted of PMVEC cultured under similar condition without clinorotation for 12 and 48 hours. Gfiess assay was performed to detect the release of nitric oxide （NO）, transmission electron microscope was used to observe the ultrastructural changes in eaveolae, and Western blotting was performed to detect the expression of eNOS and eaveolirvl. Results The production of NO and expression of eNOS were higher in microgravity- exposed 12h group and microgravity-exposed 24h group than in control 12h group and control 24h group, respectively （P〈0. 05）. By using the transmission electron microscope, caveolae, in clear feature resembling Ω, were found abundantly on cell membrane in control 12h group and control 24h group, whereas less caveolae, with obscure structure, were found in microgravity-exposed 12h group, and eaveolae were hardly found in microgravity-exposed 24h group. Western blotting showed a significant decrease in the expression of caveolin-1 in microgravity-exposed 12h group and microgravity exposed 24h group compared with control 12h group and control 24h group, respectively （P〈0. 05 or P〈0. 01）. Conclusion Simulated microgravity induced by clinorotation may inhibit the expression of caveolin-1, and increase the eNOS expression and NO production in PMVECs.

关 键 词：失重模拟 内皮细胞 毛细血管 胞膜窖 窖蛋白1 一氧化氮合酶Ⅲ型 

分 类 号：R852.22[医药卫生—航空、航天与航海医学]