模拟微重力对NIH3T3细胞近日节律基因表达的影响*

杨云云杨淑红刘延友成姝婷后望李世平王正荣

(四川大学华西基础医学与法医学院·卫生部时间生物学重点实验室，四川 成都 610041)

【摘要】目的探讨模拟微重力对NIH3T3细胞近日节律基因Per1, Per2与Cry1 mRNA表达的影响。方法体外培养培养NIH3T3细胞分为实验组和对照组。实验组采用RCCS模拟微重力环境培养NIH3T3细胞；而对照组在正常重力环境下静置培养。两组均采用PMA刺激细胞，RT-qPCR检测Per1，Per2与Cry1 mRNA的表达水平。结果对照组与实验组的近日节律基因Per1，Per2与Cry1的mRNA都呈现节律性。与对照相比，实验组节律基因的振幅减小，相位延迟。结论模拟微重力影响NIH3T3细胞近日节律基因Per1，Per2，Cry1的表达。

【关键词】模拟微重力； 近日节律基因； NIH3T3；

【中图分类号】R 318.01【文献标志码】A

基金项目：国家重点基础研究发展规划项目(2011CB71 1000)

通讯作者：刘延友，E-mail：liuyanyou@126.com

收稿日期：( 2015-01-30； 编辑： 张文秀)

The effect of stimulated microgravity on expression of circadian genesYANG Yunyun, YANG Shuhong, LIU Yanyou,etal

(KeyLaboratoryofChronobiology,MinistryofHealth(SichuanUniversity),WestChinaSchoolofPreclinicaland

ForensicMedicine,SichuanUniversity,Chengdu610041,China)

Abstract【】ObjectiveTo investigate the effect of simulated microgravity on circadian genes in NIH3T3 cells. MethodsNIH3T3 cells cultured in vitro were divided into control group and experiment group. The experiment group was cultured in Rotary Cell Culture System (RCCS) stimulated microgravity. The control group was cultured under static condition. Both two groups were stimulated by PMA. The expression of Per1, Per2 and Cry1 were detected by RT-qPCR. ResultsThe mRNA level of Per1, Per2 and Cry1 demonstrated rhythmic oscillation in control group and experiment group. The amplitudes of Per1, Per2 and Cry1 of experiment group were lower than that of control group. The phases of Per1, Per2 and Cry1 of experiment group delayed compared that of control group. ConclusionSimulated microgravity can influence the expression of Per1, Per2 and Cry1.

【Key words】Stimulated microgravity; Circadian genes; NIH3T3

近日节律广泛存在于生物界。在自然状态下，生物钟接受外界光-暗和温度等周期信号，调整自身的相位，与外界环境保持同步[1]。生物节律由近日节律基因的转录-翻译的正负反馈环路产生[2，3]。生物体内约有10%的基因参与调节近日节律[4，5]。目前，已通过基因分析找到与近日节律有关的节律基因，如Perl、Per2、Per3、Clock、Bmal、Tim、Cryl、Cry2和Tau等，其中Per1，Per2，Cry1为近日节律的核心元件。

随着航天事业的发展，微重力对有机体的影响成为研究热点。微重力可以破坏细胞的骨架，改变细胞的形态，使细胞发生水肿，微管蛋白减少，部分细胞发生凋亡等[6，7]。在微重力条件下，人T淋巴细胞减少，免疫因子的表达出现明显的下降。……