【摘要】：背景與目的 生物節(jié)律是生物體在進(jìn)化過(guò)程中為抵御大自然環(huán)境而逐漸形成機(jī)體內(nèi)在與大自然環(huán)境周期性變化相似的生物節(jié)律。生物節(jié)律的中樞起搏點(diǎn)位于下丘腦視交叉上核，視交叉上核將接收到的信號(hào)通過(guò)調(diào)節(jié)激素及體液等進(jìn)而將信號(hào)傳輸?shù)酵庵芙M織肝、腎、脾等，從而使外周組織呈節(jié)律性震蕩。近年來(lái)有研究表明生物節(jié)律在抗腫瘤治療、調(diào)節(jié)睡眠以及提高航天員工作績(jī)效方面有重要作用。本研究通過(guò)在全黑環(huán)境條件下對(duì)C57小鼠進(jìn)行后肢去負(fù)荷運(yùn)動(dòng)(模擬微重力的尾吊)的方法，檢測(cè)小鼠血漿中褪黑素與皮質(zhì)酮激素水平及肝臟組織中節(jié)律基因clock、bmal1、per1、per2、cry1、cry2的變化，探究生物節(jié)律與微重力的關(guān)聯(lián)性。 方法 常規(guī)明暗光照條件導(dǎo)引C57小鼠一周后，在持續(xù)黑暗條件下采用頭低位-30度法尾吊C57小鼠，在尾吊小鼠第12天、13天、14天每隔4小時(shí)連續(xù)18個(gè)時(shí)間點(diǎn)進(jìn)行血漿與肝臟組織取材。選取外周血血漿采用酶聯(lián)免疫吸附法(ELISA)檢測(cè)褪黑素與皮質(zhì)酮激素濃度，選取肝臟組織提取總RNA，用Real-time PCR法檢測(cè)節(jié)律基因(clock、bmal1、per1、per2、cry1、cry2) mRNA的相對(duì)表達(dá)水平。 結(jié)果 (1)全黑環(huán)境條件下，C57小鼠尾吊組與對(duì)照組血漿中褪黑素與皮質(zhì)酮激素濃度無(wú)顯著性差異(P0.05)。 (2)尾吊組與對(duì)照組肝臟組織中clock與bmal1節(jié)律基因mRNA水平均呈現(xiàn)出節(jié)律性表達(dá)特征。在尾吊條件下，clock基因mRNA的峰值相位提前約4 h，bmal1基因mRNA表達(dá)節(jié)律的振幅在每天授時(shí)因子時(shí)間ZT2(Zeitgeber Time)與ZT6顯著下降(P0.05)。 (3)尾吊組與對(duì)照組肝臟中per1與per2節(jié)律基因的mRNA表達(dá)峰值均為約下午14-18時(shí)，谷值約為上午10時(shí)。 (4)尾吊組與對(duì)照組肝臟中cry1與cry2節(jié)律基因的mRNA的表達(dá)水平呈現(xiàn)相同的趨勢(shì)。峰值均約為午后14時(shí)，谷值約為早晨6時(shí)。 結(jié)論 在持續(xù)全黑暗條件下，C57小鼠肝臟組織節(jié)律基因clock與bmal1的mRNA水平表現(xiàn)出明顯的近日節(jié)律性，尾吊(后肢去負(fù)荷運(yùn)動(dòng))對(duì)clock基因產(chǎn)生峰值相位提前的效應(yīng)，對(duì)bmal1基因產(chǎn)生振幅減少的效應(yīng)；節(jié)律基因per1與per2、cry1與cry2的表達(dá)分別存在同步化的特征。
[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.
【學(xué)位授予單位】：中北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：G804.22

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