小鼠外周血白细胞节律相关基因cry2,per2,timeless,rev-erb表达的近日节律性研究  

Circadian expressions of related genes of cry2,per2,timeless and rev-erb in peripheral white blood cells in mice

作　　者：李科华[1] 王庆敏[1] 刘秋红[1] 戴圣龙[1] 时粉周[1] 姚永杰[1] 

机构地区：[1]海军医学研究所航空医学研究室,上海200433

出　　处：《海军医学杂志》2014年第3期169-173,共5页Journal of Navy Medicine

基　　金：总后卫生部面上项目资助(CWS11J029);海军医学研究所所基金资助(10HY51)

摘　　要：目的探讨在12 h光照/12 h黑暗(12L/12D)光暗循环条件下,小鼠外周血白细胞中相关基因cry2,per2,timeless,rev-erb近日节律性表达特点。方法用完全随机分组法将C57/BL6小鼠分成6组,每组对应一个时相点,在12L/12D条件下词养4周后分别在6个时相点(9:00,13:00,17:00,21:00,1:00,5:00)取外周血并分离白细胞。抽提总RNA,采用实时荧光定量PCR技术检测小鼠外周血白细胞cry2,per2,timeless,rev-erb 4种节律基因的mRNA水平,用余弦函数拟合节律参数,并经振幅f检验分析是否存在近日节律。结果 cry2,per2,timeless,rev-erb 4种基因mRNA存在明显的近日节律性,各基因的峰值明显高于谷值(P<0.01);per2峰值相位时间在4:00左右,而cry2,timeless,rev-erb基因的峰值时间在11:00-12:00;从峰值时间和震荡幅度看,cry2,timeless,rev-erb峰值时间比per2超前,振幅比per2高,cry2、timeless、rev-erb的峰值时间、振幅大小比较接近。结论 C57/BL6小鼠外周血白细胞4种基因cry2,per2,timeless,rev-erb的mRNA存在明显的近日节律性。cry2,timeless,rev-erb在负反馈中的作用强于per2,而cry2,timeless,rev-erb在负反馈中的作用相似。该研究为深入理解哺乳动物的免疫功能昼夜波动打下了基础,也为诊断、治疗免疫功能紊乱相关疾病提供了靶目标。但这4种基因如何调控外周血中的免疫细胞,尚需进一步深入研究。Objective To investigate features of circadian expressions of related genes of cry2,per2,timeless and rev-erb in peripheral white blood cells in mice under the 12 h-light and 12 h-dark cycle conditions. Methods Forty-eight male C57 /BL6 mice were randomly divided into 6 groups,with each group directed to a corresponding time point. The animals were kept under the light regimen of LD cycle（ 12L /12D） for four weeks. Then,peripheral blood samples were taken at 6 time points（ 9： 00,13： 00,17： 00,21： 00,1： 00,5： 00） and white blood cells were separated. Total RNA was extracted from the white blood cells and mRNA levels of each gene（ cry2,per2,timeless and rev-erb） were detected by reverse transcription polymerase chain reaction（ real time PCR） at different time points. The data were analyzed both by the cosine function software and the amplitude F test to investigate the circadian rhythm. Results Under the light regimen of LD cycle,the four genes in white blood cells showed a significant circadian oscillation,with the apexes of different genes being obviously higher than the troughs（ P〈0. 01）. The apex of per2 was at about 4： 00,while the apex of the other three genes were at about 11： 00-12： 00. From the standpoints of apex time and oscillation,the apex time of cry2,timeless and rev-erb was before that of per2,with a higher level of amplitude than that of per2. The apex time of cry2,timeless,rev-erb mRNA was before that of per2,with the amplitude being higher than that of per2. However,the mRNAs of cry2,timeless and rev-erb showed a similar apex time and amplitude. Conclusion Research findings demonstrated that the expressions of the 4 genes in peripheral white blood cells showed an obvious circadian rhythm. It seemed that the effects of cry2,timeless and rev-erb were stronger than that of per2 in the negative regulation,while the effect of rev-erb was similar in the negative regulation. This research might not only be helpful to the deep understanding of the circadian fluctua

关 键 词：近日节律 基因 实时定量PCR 

分 类 号：R392.6[医药卫生—免疫学]