后肢去负荷运动对小鼠生物节律的影响

发布时间：2018-11-11 07:16

【摘要】：背景与目的 生物节律是生物体在进化过程中为抵御大自然环境而逐渐形成机体内在与大自然环境周期性变化相似的生物节律。生物节律的中枢起搏点位于下丘脑视交叉上核，视交叉上核将接收到的信号通过调节激素及体液等进而将信号传输到外周组织肝、肾、脾等，从而使外周组织呈节律性震荡。近年来有研究表明生物节律在抗肿瘤治疗、调节睡眠以及提高航天员工作绩效方面有重要作用。本研究通过在全黑环境条件下对C57小鼠进行后肢去负荷运动(模拟微重力的尾吊)的方法，检测小鼠血浆中褪黑素与皮质酮激素水平及肝脏组织中节律基因clock、bmal1、per1、per2、cry1、cry2的变化，探究生物节律与微重力的关联性。 方法 常规明暗光照条件导引C57小鼠一周后，在持续黑暗条件下采用头低位-30度法尾吊C57小鼠，在尾吊小鼠第12天、13天、14天每隔4小时连续18个时间点进行血浆与肝脏组织取材。选取外周血血浆采用酶联免疫吸附法(ELISA)检测褪黑素与皮质酮激素浓度，选取肝脏组织提取总RNA，用Real-time PCR法检测节律基因(clock、bmal1、per1、per2、cry1、cry2) mRNA的相对表达水平。 结果 (1)全黑环境条件下，C57小鼠尾吊组与对照组血浆中褪黑素与皮质酮激素浓度无显著性差异(P0.05)。 (2)尾吊组与对照组肝脏组织中clock与bmal1节律基因mRNA水平均呈现出节律性表达特征。在尾吊条件下，clock基因mRNA的峰值相位提前约4 h，bmal1基因mRNA表达节律的振幅在每天授时因子时间ZT2(Zeitgeber Time)与ZT6显著下降(P0.05)。 (3)尾吊组与对照组肝脏中per1与per2节律基因的mRNA表达峰值均为约下午14-18时，谷值约为上午10时。 (4)尾吊组与对照组肝脏中cry1与cry2节律基因的mRNA的表达水平呈现相同的趋势。峰值均约为午后14时，谷值约为早晨6时。 结论 在持续全黑暗条件下，C57小鼠肝脏组织节律基因clock与bmal1的mRNA水平表现出明显的近日节律性，尾吊(后肢去负荷运动)对clock基因产生峰值相位提前的效应，对bmal1基因产生振幅减少的效应；节律基因per1与per2、cry1与cry2的表达分别存在同步化的特征。
[Abstract]:Background and objective Biological rhythms are biological rhythms that are similar to the periodic changes in natural environments in order to resist the natural environment during the evolution of organisms. The central pacing point of biological rhythm is located in the supraoptic nucleus of hypothalamus. The signal is transmitted to the peripheral liver, kidney, spleen and so on by regulating hormones and body fluids. Recent studies have shown that biological rhythms play an important role in antitumor therapy, sleep regulation and performance improvement. The aim of this study was to detect the levels of melatonin and corticosterone in plasma and clock,bmal1,per1,per2, in liver tissue of C57 mice by means of hind limb deloading (simulated microgravity tail suspension) under the condition of full black environment. Changes in cry1,cry2, exploring the relationship between biological rhythms and microgravity. Methods C57 mice were exposed to conventional light and dark illumination for one week. C57 mice were suspended by head-low 30 degree tail suspension under the condition of continuous darkness. The mice were suspended on the 12th and 13th day of tail suspension. Plasma and liver tissue were collected from the liver at 18 consecutive time points every 4 hours on 14 days. The concentrations of melatonin and corticosterone in peripheral blood plasma were detected by enzyme linked immunosorbent assay (ELISA) and total RNA, was extracted from liver tissue to detect the rhythm gene (clock,bmal1,per1,per2,cry1,) by Real-time PCR method. Cry2) the relative expression level of mRNA. Results (1) there was no significant difference in the concentrations of melatonin and corticosterone between the tail suspension group and the control group (P0.05). (2) the expression of clock and bmal1 rhythm gene mRNA in liver tissues of tail suspension group and control group were rhythmic. Under the condition of tail suspension, the amplitude of mRNA expression rhythm of clock gene mRNA was significantly decreased at the time of daily timing factor ZT2 (Zeitgeber Time) and ZT6 (P0.05). (3) the peak value of mRNA expression of per1 and per2 rhythm genes in the liver of tail suspension group and control group was about 14-18 pm, and the valley value was about 10:00 in the morning. (4) the level of mRNA expression of cry1 and cry2 rhythm gene in the liver of tail suspension group and control group showed the same trend. The peak value is about 14:00 in the afternoon and the valley value is about 6 in the morning. Conclusion under the condition of continuous complete darkness, the mRNA levels of clock and bmal1 in liver tissue of C57 mice showed an obvious circadian rhythm, and tail suspension (hind limb deloading) had a prematurely peak phase effect on clock gene. The effect of amplitude reduction on bmal1 gene was observed. The expression of rhythm gene per1, per2,cry1 and cry2 were synchronized.
【学位授予单位】：中北大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：G804.22

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