模拟微重力对NIH-3T3细胞clock及bmal1基因表达的影响  

Effects of Simulated Microgravity on Clock and Bmal1 Gene Expression in NIH-3T3 Cells

作　　者：王艳利[1] 吕柯[2] 钟悦 江舟[4] 曲丽娜[2] 陈海龙[2] 毕蕾[2] 曹宏卿[2] 李秀锦[1] 李莹辉[2] 

机构地区：[1]燕山大学环境与化学工程学院,河北秦皇岛066004 [2]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,北京100094 [3]航天城门诊部,北京100094 [4]四川大学华西基础医学与法医学院,四川成都610041

出　　处：《航天医学与医学工程》2012年第1期18-21,共4页Space Medicine & Medical Engineering

基　　金：航天医学基础与应用国家重点实验室自主项目(SMFA09A06);中国载人航天预先研究项目(SJ200908);国家重点基础研究计划资助项目(2011CB711000)

摘　　要：目的探讨模拟微重力对NIH-3T3细胞节律基因clock与bmal1 mRNA表达的影响。方法 NIH-3T3细胞采用回转器分别回转培养6 h,12 h,18 h,24 h,30 h,36 h,42 h和48 h,同时设置1 G静置培养为对照组。Trizol试剂提取细胞总RNA,实时定量PCR法检测节律基因clock及bmal1 mRNA在各时间点的相对表达量。通过余弦法获取节律参数,并经振幅检验分析是否存在昼夜节律。结果 clock及bmal1基因mRNA的表达趋势相近。对照组clock与bmal1 mRNA相对水平呈现节律性表达,而回转组表达无节律性。回转组mRNA相对水平最大值出现于约6 h时间点,最小值出现在约18 h,而对照组分别出现于约18 h与24 h。与对照组比回转组的周期延长了约16 h。回转后clock与bmal1的峰值相位前移。结论 NIH-3T3细胞中clock与bmal1的表达存在同步化的转录特征。模拟微重力可影响clock与bmal1节律的周期长度及峰值相位。Objective To investigate the effects of simulated microgravity on clock and bmal1 gene expression in NIH-3T3 cells.Methods NIH-3T3 fibroblast cells were used as a model of simulated microgravity with MG3 clinostat for 6 h,12 h,18 h,24 h,30 h,36 h,42 h and 48 h.Those cultured in 1 G condition served as group control.Cells were harvested and lysated with Trizol reagent for total cellular RNA extraction.Then RT-qPCR was used to analyze the relative mRNA expression levels of clcok and bmal1,at different time points.The circadian parameters were obtained and analyzed by the cosine function to verify the existence of circadian rhythmicity.Results The similar tendencies were found in both clock and bmal1 gene relative expression level.Rhythmicity of both clock and bmal1 mRNA expression level were identified in group control.While,such characteristics disappeared in group clinorotation.The maximum and minimum value of mRNA relative expression levels of clock and bmal1 during clinorotation were both found approximately at the time points 6 h and 18 h,respectively.Accordingly,the time points of the group control were 18 h and 24 h.The period length of group experiment was about 16 h longer than control group.The peak phase and peak time of clock and bmal1 with group simulated weightlessness were shifted ahead of group control.Conclusion The expression of clock and bmal1 in the NIH-3T3 cells display a synchronous circadian transcription characteristics.The period length,peak phase of circadian rhythms of clock and bmal1 mRNA level could be significantly altered by simulated weightlessness.

关 键 词：模拟微重力 近日节律 CLOCK基因 bmal1基因 

分 类 号：R852.6[医药卫生—航空、航天与航海医学]