背景24小時(shí)生物節(jié)律控制生物進(jìn)程的多樣性。系統(tǒng)生物鐘位于大腦的視交叉神經(jīng)核,在很多的外周器官和組織中都表現(xiàn)出生物節(jié)律變化,例如:肝臟、肺、腎臟、心臟和皮膚。在分子水平上,生物節(jié)律的中心調(diào)控由4個(gè)重要的核心生物鐘蛋白組成,包括brain and muscle ARNT-like(BMAL1),circadian locomotor output cycles kaput(CLOCK),period(PERs)和cryptochrome(CRYs)。這4種轉(zhuǎn)錄因子的表達(dá)通過(guò)轉(zhuǎn)錄翻譯反饋回路調(diào)節(jié)。其中BMAL1和CLOCK形成異質(zhì)二聚體并通過(guò)結(jié)合在其下游基因啟動(dòng)子區(qū)域的E-box(CACGTG)序列來(lái)正向調(diào)節(jié)基因表達(dá),從而作為生物鐘蛋白中正向分枝來(lái)起作用。CRYs和PERs則相反,它們屬于BMAL-CLOCK的下游基因。當(dāng)其表達(dá)積累到一定水平后,二者形成異質(zhì)二聚體,并由胞漿移動(dòng)至細(xì)胞核內(nèi),從而抑制BMAL-CLOCK的表達(dá),產(chǎn)生所謂的負(fù)反饋調(diào)節(jié)回路。除此之外,BMAL1的表達(dá)還受另外兩個(gè)孤核受體RORs和REV-ERBs的調(diào)節(jié)。RORs能夠促進(jìn)BMAL1的表達(dá),被認(rèn)為是生物節(jié)律的正向調(diào)節(jié)因子...

【文章頁(yè)數(shù)】：115 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
摘要
abstract
Chapter1 Introduction
    1.1 Introduction of Human Skin
        1.1.1 Introduction of the morphology and functions of human skin
    1.2 Introduction of the Circadian Rhythm in the skin
        1.2.1 The molecular components of circadian clock
        1.2.2 Circadian Clock biological functions in skin
        1.2.3 Skin diseases related to abnormal circadian rhythm
    1.3 Circadian rhythm controls UV-induced DNA damage in the skin
        1.3.1 UV Rays in the skin
        1.3.2 UV-induced DNA damage in the skin
        1.3.3 Circadian clock and UV-induced DNA damage
        1.3.4 Circadian clock and DNA damage repair response
    1.4 Retinoid-related orphan nuclear receptorα,RORα
        1.4.1 Structure characteristic of RORα
        1.4.2 Biological functions of RORα
    1.5 RORαand Circadian clock
    1.6 Hypothesis and Aims
Chapter2 Experimental Materials and Methods
    2.1 Experiment materials
        2.1.1 Cell lines
        2.1.2 Antibody and reagent
        2.1.3 Preparation of the solutions
        2.1.4 Instruments and equipment
    2.2 Method
        2.2.1 Cell culture
        2.2.2 Small RNA interference the expression of Circadian Clock genes Bmal1 and Clock in cells
        2.2.3 Using shRNAs to establish RORαknockout stable cell line
        2.2.4 UVB irradiation
        2.2.5 Real time PCR for cellular mRNA levels detection
        2.2.6 BCA Protein Quantification
        2.2.7 Western Blot for cellular protein level detection
        2.2.8 Immunofluorescence Microscopy detect target genes expression in cells
        2.2.9 Cell viability assay
        2.2.10 Cellular Apoptosis detection
        2.2.11 Data analysis
Chapter3 Result
    3.1 Alterations in the expression of circadian clock genes in the immortal human adult low calcium temperature(HaCaT)keratinocytes following low-dose UVB exposure
        3.1.1 Low dose of UVB induces cycling of Circadian Clock genes in Hacat keratinocytes
        3.1.2 Low dose of UVB induces cycling of cytokines release in Hacat keratinocytes
    3.2 Circadian Clock genes are essential for regulates HaCat keratinocytes Proliferation under UVB irradiation condition
        3.2.1 BMAL1 and CLOCK proteins are critical for regulates HaCat keratinocytes Proliferation
    3.3 Clock genes are essential for regulates HaCat keratinocytes Apoptosis under UVB exposure condition
        3.3.1 The Core Circadian Clock genes BMAL1 and CLOCK are critical for UVB-induced HaCat keratinocytes Apoptosis
    3.4 The molecular mechanisms of clock genes regulating UVB induced HaCat keratinocytes DNA damage responses
        3.4.1 BMAL1 and CLOCK are not indispensable for UVB-induced CPDs formation. ..
        3.4.2 BMAL1 and CLOCK are not involved in activation of DNA damage checkpoints in HaCat keratinocytes
        3.4.3 BMAL1 and CLOCK are involved in activation of histone H2A.X in HaCat keratinocytes
    3.5 BMAL1 and CLOCK control primary human keratinocytes proliferation and apoptosis under UVB irradiation condition
        3.5.1 BMAL1 and CLOCK control primary human keratinocytes growth under5 m J/cm2 of UVB irradiation
        3.5.2 BMAL1 and CLOCK control primary human keratinocytes apoptosis under low dose of UVB irradiation
    3.6 Effects of BMAL1 and CLOCK depletion on UVB-induced DNA damage responses in primary human keratinocytes
        3.6.1 BMAL1 or CLOCK depletion has no effect on UVB-induced DNA damage responses at early time
        3.6.2 CLOCK protein depletion attenuates UVB-induced p53 activity in primary human keratinocytes
    3.7 Depletion of BMAL1 or CLOCK triggers terminal differentiation in HKCs
    3.8 Clock genes are essential for regulating cytokines release under UVB irradiation in HaCat keratinocytes
        3.8.1 BMAL1 and CLOCK control UVB induced cytokines release in HaCat cells. ..
    3.9 RORA is essential for UVB-induced Apoptosis in HaCat keratinocytes
    3.10 RORA is critical for UVB induced cytokines release in HaCat cells
        3.10.1 RORA knockdown receded UVB-induced cytokines release in HaCat cells
        3.10.2 Overexpression of RORA slightly increased UVB-induced cytokines release in HaCat cells
    3.11 RORA controls the core clock gene BMAL1 expression under UVB irradiation in HaCat keratinocytes
Chapter4 Discussion and Prospect
    4.1 Discussion and Conclusion
        4.1.1 Discussion
        4.1.2 Conclusion
    4.2 Prospect
References
Published papers
Scientific research
Acknowledgement



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