| 摘要 | 第4-11页 |
| abstract | 第11-12页 |
| Chapter1 Introduction | 第17-37页 |
| 1.1 Introduction of Human Skin | 第17-18页 |
| 1.1.1 Introduction of the morphology and functions of human skin | 第17-18页 |
| 1.2 Introduction of the Circadian Rhythm in the skin | 第18-25页 |
| 1.2.1 The molecular components of circadian clock | 第19-21页 |
| 1.2.2 Circadian Clock biological functions in skin | 第21-24页 |
| 1.2.3 Skin diseases related to abnormal circadian rhythm | 第24-25页 |
| 1.3 Circadian rhythm controls UV-induced DNA damage in the skin | 第25-30页 |
| 1.3.1 UV Rays in the skin | 第25-26页 |
| 1.3.2 UV-induced DNA damage in the skin | 第26-28页 |
| 1.3.3 Circadian clock and UV-induced DNA damage | 第28-29页 |
| 1.3.4 Circadian clock and DNA damage repair response | 第29-30页 |
| 1.4 Retinoid-related orphan nuclear receptorα,RORα | 第30-34页 |
| 1.4.1 Structure characteristic of RORα | 第30-32页 |
| 1.4.2 Biological functions of RORα | 第32-34页 |
| 1.5 RORαand Circadian clock | 第34-35页 |
| 1.6 Hypothesis and Aims | 第35-37页 |
| Chapter2 Experimental Materials and Methods | 第37-63页 |
| 2.1 Experiment materials | 第37-42页 |
| 2.1.1 Cell lines | 第37页 |
| 2.1.2 Antibody and reagent | 第37-39页 |
| 2.1.3 Preparation of the solutions | 第39-40页 |
| 2.1.4 Instruments and equipment | 第40-42页 |
| 2.2 Method | 第42-63页 |
| 2.2.1 Cell culture | 第42-43页 |
| 2.2.2 Small RNA interference the expression of Circadian Clock genes Bmal1 and Clock in cells | 第43-44页 |
| 2.2.3 Using shRNAs to establish RORαknockout stable cell line | 第44-48页 |
| 2.2.4 UVB irradiation | 第48页 |
| 2.2.5 Real time PCR for cellular mRNA levels detection | 第48-53页 |
| 2.2.6 BCA Protein Quantification | 第53-54页 |
| 2.2.7 Western Blot for cellular protein level detection | 第54-57页 |
| 2.2.8 Immunofluorescence Microscopy detect target genes expression in cells | 第57-58页 |
| 2.2.9 Cell viability assay | 第58-59页 |
| 2.2.10 Cellular Apoptosis detection | 第59-61页 |
| 2.2.11 Data analysis | 第61-63页 |
| Chapter3 Result | 第63-93页 |
| 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 | 第63-65页 |
| 3.1.1 Low dose of UVB induces cycling of Circadian Clock genes in Hacat keratinocytes | 第63-64页 |
| 3.1.2 Low dose of UVB induces cycling of cytokines release in Hacat keratinocytes | 第64-65页 |
| 3.2 Circadian Clock genes are essential for regulates HaCat keratinocytes Proliferation under UVB irradiation condition | 第65-69页 |
| 3.2.1 BMAL1 and CLOCK proteins are critical for regulates HaCat keratinocytes Proliferation | 第66-69页 |
| 3.3 Clock genes are essential for regulates HaCat keratinocytes Apoptosis under UVB exposure condition | 第69-71页 |
| 3.3.1 The Core Circadian Clock genes BMAL1 and CLOCK are critical for UVB-induced HaCat keratinocytes Apoptosis | 第69-71页 |
| 3.4 The molecular mechanisms of clock genes regulating UVB induced HaCat keratinocytes DNA damage responses | 第71-77页 |
| 3.4.1 BMAL1 and CLOCK are not indispensable for UVB-induced CPDs formation. .. | 第71-72页 |
| 3.4.2 BMAL1 and CLOCK are not involved in activation of DNA damage checkpoints in HaCat keratinocytes | 第72-75页 |
| 3.4.3 BMAL1 and CLOCK are involved in activation of histone H2A.X in HaCat keratinocytes | 第75-77页 |
| 3.5 BMAL1 and CLOCK control primary human keratinocytes proliferation and apoptosis under UVB irradiation condition | 第77-79页 |
| 3.5.1 BMAL1 and CLOCK control primary human keratinocytes growth under5 m J/cm2 of UVB irradiation | 第77-78页 |
| 3.5.2 BMAL1 and CLOCK control primary human keratinocytes apoptosis under low dose of UVB irradiation | 第78-79页 |
| 3.6 Effects of BMAL1 and CLOCK depletion on UVB-induced DNA damage responses in primary human keratinocytes | 第79-81页 |
| 3.6.1 BMAL1 or CLOCK depletion has no effect on UVB-induced DNA damage responses at early time | 第79-80页 |
| 3.6.2 CLOCK protein depletion attenuates UVB-induced p53 activity in primary human keratinocytes | 第80-81页 |
| 3.7 Depletion of BMAL1 or CLOCK triggers terminal differentiation in HKCs | 第81-82页 |
| 3.8 Clock genes are essential for regulating cytokines release under UVB irradiation in HaCat keratinocytes | 第82-85页 |
| 3.8.1 BMAL1 and CLOCK control UVB induced cytokines release in HaCat cells. .. | 第83-85页 |
| 3.9 RORA is essential for UVB-induced Apoptosis in HaCat keratinocytes | 第85-87页 |
| 3.10 RORA is critical for UVB induced cytokines release in HaCat cells | 第87-91页 |
| 3.10.1 RORA knockdown receded UVB-induced cytokines release in HaCat cells | 第87-89页 |
| 3.10.2 Overexpression of RORA slightly increased UVB-induced cytokines release in HaCat cells | 第89-91页 |
| 3.11 RORA controls the core clock gene BMAL1 expression under UVB irradiation in HaCat keratinocytes | 第91-93页 |
| Chapter4 Discussion and Prospect | 第93-101页 |
| 4.1 Discussion and Conclusion | 第93-98页 |
| 4.1.1 Discussion | 第93-98页 |
| 4.1.2 Conclusion | 第98页 |
| 4.2 Prospect | 第98-101页 |
| References | 第101-113页 |
| Published papers | 第113页 |
| Scientific research | 第113-115页 |
| Acknowledgement | 第115页 |
