| Acknowledgements | 第8-15页 |
| List of Symbols and Abbreviations | 第15-17页 |
| Abstract | 第17-23页 |
| Abstract in Chinese(概要) | 第24-30页 |
| Introduction | 第30-33页 |
| Chapter 1 TIGAR is a sensitive biosensor for tight junction injury during low-glucose stress | 第33-45页 |
| 1.1 Background | 第34页 |
| 1.2 Material and methods | 第34-37页 |
| 1.2.1 Reagents | 第34页 |
| 1.2.2 Low-glucose stress induction and Experimental treatments of cellCultures | 第34-35页 |
| 1.2.3 Plasmid Construction and Transfection | 第35页 |
| 1.2.4 Lentiviral transfection | 第35页 |
| 1.2.5 Cell Culture TAT induction | 第35页 |
| 1.2.6 Cell fractionation and Western blot assay | 第35-36页 |
| 1.2.7 Confocal immunofluorescence staining and analysis | 第36页 |
| 1.2.8 Transmission Electron Microscopy | 第36-37页 |
| 1.2.9 Statistical analysis | 第37页 |
| 1.3 Results | 第37-43页 |
| 1.3.1 TIGAR is dysregulated during low-glucose stimulation | 第37页 |
| 1.3.2 Temporal changes in tight junction proteins, stress and energyhomeostasis biomarkers | 第37-40页 |
| 1.3.3 TIGAR is a sensitive stress monitor reflects tight junction injury uponLow-glucose treatment | 第40页 |
| 1.3.4 Protective role of TIGAR on tight junction integrity during low-glucoseInduced stress | 第40-43页 |
| 1.4 Discussion | 第43-44页 |
| 1.5 Summary | 第44-45页 |
| Chapter 2 TIGAR regulates autophagy during low-glucose induced insult | 第45-58页 |
| 2.1 Background | 第46页 |
| 2.2 Material and methods | 第46-49页 |
| 2.2.1 Reagents | 第46页 |
| 2.2.2 Low-glucose stress induction and experimental treatments of cellcultures | 第46-47页 |
| 2.2.3 RNA interference | 第47页 |
| 2.2.4 Immunofluorescent assay | 第47页 |
| 2.2.5 Cell Culture TAT induction | 第47-48页 |
| 2.2.6 Lentiviral transfection | 第48页 |
| 2.2.7 Cell fractionation and Western blot assay | 第48页 |
| 2.2.8 Transmission Electron Microscopy | 第48-49页 |
| 2.2.9 Statistical analysis | 第49页 |
| 2.3 Results | 第49-56页 |
| 2.3.1 Disturbance of autophagic flux during low-glucose stress | 第49-51页 |
| 2.3.2 TIGAR regulates autophagic signaling during low-glucose induced stress | 第51-54页 |
| 2.3.3 Protective role of TIGAR through autophagy inhibitionduring low-glucose stress | 第54页 |
| 2.3.4 Pharmacological inhibition of TIGAR increasesautophagosome formation | 第54-56页 |
| 2.4 Discussion | 第56-57页 |
| 2.5 Summary | 第57-58页 |
| Chapter 3 Protein tyrosine nitration of TIGAR during Hypoglycemia | 第58-78页 |
| 3.1 Background | 第59页 |
| 3.2 Material and methods | 第59-63页 |
| 3.2.1 Reagents | 第59页 |
| 3.2.2 Low-glucose stress induction and treatments | 第59-60页 |
| 3.2.3 Plasmid Construction and Transfection | 第60页 |
| 3.2.4 Lentiviral transfection | 第60页 |
| 3.2.5 Immunofluorescence staining and analysis | 第60-61页 |
| 3.2.6 Cell fractionation, immunoprecipitation and immunoblot analysis | 第61-62页 |
| 3.2.7 Fluorescence Resonance Energy Transfer | 第62页 |
| 3.2.8 In vitro protein nitration and Mass spectrometric assay | 第62-63页 |
| 3.2.9 Measurements of NADPH levels | 第63页 |
| 3.2.10 Statistical analysis | 第63页 |
| 3.3 Results | 第63-76页 |
| 3.3.1 Nitrosative stress initiated TIGAR dysfunction during low-glucosemediated stress | 第63-65页 |
| 3.3.2 Identification of Tyrosine 92 as TIGAR nitrated site | 第65页 |
| 3.3.3 TIGAR is not associated with HKII during low-glucose induced stress | 第65-66页 |
| 3.3.4 Identification of differentially expressed binding partners of TIGAR | 第66-69页 |
| 3.3.5 Calmodulin interacts with TIGAR | 第69-72页 |
| 3.3.6 Mutation of Tyrosine 92 abolished the protective effect of TIGAR | 第72-74页 |
| 3.3.7 Correlation of Tyrosine 92 nitration of TIGAR with autophagy activationin brain endothelial cell | 第74-76页 |
| 3.4 Discussion | 第76-77页 |
| 3.5 Summary | 第77-78页 |
| Chapter4 Neurovascular protective role of TIGAR during insulin induced hypoglycemia inmice | 第78-86页 |
| 4.1 Background | 第79页 |
| 4.2 Material and methods | 第79-81页 |
| 4.2.1 Reagents | 第79页 |
| 4.2.2 Experimental animals | 第79-80页 |
| 4.2.3 Mice model of Insulin induced hypoglycemia | 第80页 |
| 4.2.4 PET-Scan data and analysis | 第80页 |
| 4.2.5 Immunohistochemical analysis | 第80-81页 |
| 4.2.6 Transmission Electron Microscopy | 第81页 |
| 4.2.7 Statistical analysis | 第81页 |
| 4.3 Results | 第81-84页 |
| 4.3.1 PET Scanning of mice model of insulin induced hypoglycemia | 第81-82页 |
| 4.3.2 Vasoprotective role of TIGAR during hypoglycemia induced stress inmice | 第82-84页 |
| 4.4 Discussion | 第84-85页 |
| 4.5 Summary | 第85-86页 |
| Chapters 5 Illustrative Summary about the role of Endothelial TIGAR in hypoglycemia inducedstress | 第86-89页 |
| Chapter 6 Conclusions and Future perspectives | 第89-94页 |
| 6.1 Conclusions | 第90-93页 |
| 6.2 Future perspectives | 第93-94页 |
| References | 第94-103页 |
| Author biography | 第103-104页 |
| Author introduction | 第103页 |
| Honors and awards | 第103页 |
| Publications | 第103-104页 |
