| 中文摘要 | 第3-7页 |
| Abstract | 第7-11页 |
| Table of contents | 第12-15页 |
| List of Abbreviations | 第15-17页 |
| Chapter 1. A brief review about the role of PTEN-PDK1-Akt signaling inmaintaining cardiomyocytes homeostasis | 第17-33页 |
| 1. Introduction | 第18-20页 |
| 2. Akt promotes cardiomyocytes survival | 第20-21页 |
| 3. Akt facilitates cardiomyocytes proliferation | 第21页 |
| 4. Akt controls cardiomyocytes growth and size | 第21-22页 |
| 5. Akt regulates cardiomyocytes glucose metabolism | 第22-23页 |
| 6. Akt affects myocardial contractility and calcium signaling | 第23-24页 |
| 7. Akt influences cardiomyocytes mitochondrial integrity | 第24-26页 |
| 8. References | 第26-33页 |
| Chapter 2 Inactivation of PTEN reverses heart failure caused by Pdk1 deletionthrou-gh mTORC2 activation in mice | 第33-57页 |
| 1. Abstract | 第34-35页 |
| 2. Introduction | 第35-36页 |
| 3. Materials and methods | 第36-39页 |
| 3.1 Mice | 第36页 |
| 3.2 Murine Embryonic Fibroblasts (MEFs) culture | 第36-37页 |
| 3.3 Akt kinase assay | 第37页 |
| 3.4 Echocardiography | 第37页 |
| 3.5 Administration of rapamycin, PF-4708671 and bpv | 第37页 |
| 3.6 Western blot analysis | 第37-38页 |
| 3.7 Histology | 第38页 |
| 3.8 TUNEL assay | 第38页 |
| 3.9 Statistical analysis | 第38-39页 |
| 4. Results | 第39-49页 |
| 4.1 Deletion of Pten reversed heart failure caused by Pdkl deficiency | 第39-40页 |
| 4.2 Deletion of Pten enhanced Akt S473 phosphorylation in Pdk1-deficient mice | 第40-42页 |
| 4.3 Comparison of cardiomyocyte-specific,Akt1/2-deletion and Pdk1-deletion mice | 第42-44页 |
| 4.4 Akt S473 phosphorylation was both mTORC2- and PI3K-dependent in the heart | 第44-45页 |
| 4.5 Disruption of mTORC2 through deletion of Rictor in myocardium deteriorated heart failure in Pdk1-deficient mice | 第45-47页 |
| 4.6 Loss of Rictor abolished the protection against heart failure conferred by Pten inactivation in Pdk1-deficient mice | 第47页 |
| 4.7 Augment of Akt activity through chemical inhibition of PTEN and relieving S6K-Akt negative feedback prolonged the survival of Pdk1-deletion mice | 第47-49页 |
| 5. Discussion | 第49-51页 |
| 5.1 Contribution of Akt T308 and S473 phosphorylation to Akt activity and heart function | 第49-50页 |
| 5.2 PTEN-PDK1-Akt signaling regulation in heart function and heart failure | 第50页 |
| 5.3 Novel therapeutic strategies to treat human heart failure effectively | 第50-51页 |
| 6. Acknowledgements | 第51页 |
| 7. Funding | 第51-52页 |
| 8. References | 第52-57页 |
| Chapter 3. Akt regulates postnatal cardio myocytes growth and apoptosis in adosage dependent manner | 第57-71页 |
| 1. Abstract | 第58-59页 |
| 2. Introduction | 第59页 |
| 3. Methods | 第59-62页 |
| 3.1 Mice | 第59-60页 |
| 3.2 Histology and immunofluorescence staining | 第60-61页 |
| 3.3 TUNEL staining | 第61页 |
| 3.4 Protein extraction and Western blot | 第61页 |
| 3.5 Echocardiography | 第61页 |
| 3.6 Statistics | 第61-62页 |
| 4. Results | 第62-67页 |
| 4.1 Akt knockout mice have heart failure in a dosage dependent manner | 第62-63页 |
| 4.2 Akt1~(f/f);α-MHC-cre; Akt2~(-/-) mice's hearts growth are slower than control mice and gradually have cardiomyocytes apoptosis and heart failure | 第63-65页 |
| 4.3 Akt1~(f/f);α-MHC-cre; Akt2~(-/-) Akt3~(-/-) mice have more severe phenotype than Akt1~(f/f);α-MHC-cre; Akt2~(-/-) mice | 第65-66页 |
| 4.4 Many singaling pathways downstream of Akt are changed in Akt knock out mice | 第66-67页 |
| 5. Discussion | 第67-69页 |
| 5.1 Akt1 deficiency isn't enough to induce heart disfunction | 第67-68页 |
| 5.2 Heart growth retard is the initial phenotype of Akt knock out mice | 第68-69页 |
| 6. Reference | 第69-71页 |
| Chapter 4. Phosphorylation of the Twist1-family basic helix-loop-helixtranscription factors is involved in pathological cardiac remodeling | 第71-101页 |
| 1. Abstract | 第72页 |
| 2. Introduction | 第72-74页 |
| 3. Materials and Methods | 第74-81页 |
| 3.1 Chemicals and antibodies | 第74页 |
| 3.2 Plasmids construction | 第74-75页 |
| 3.3 GST-HA-mHandl fusion protein purification and generation of antibodies | 第75-76页 |
| 3.4 Cell culture, transfection, treatment and luciferase reporter assay | 第76页 |
| 3.5 Neonatal mouse cardiomyocyte culture and lenti-viral infection | 第76-77页 |
| 3.6 In vitro and in vivo phosphorylation assay | 第77页 |
| 3.7 Western blotting analysis and immunoprecipitation | 第77-78页 |
| 3.8 Gel shift assay | 第78页 |
| 3.9 ChIP assay | 第78-79页 |
| 3.10 Mice | 第79页 |
| 3.11 Echocardiography(Echo) | 第79页 |
| 3.12 Histology and immunohistochemical staining | 第79-80页 |
| 3.13 Microarray analysis | 第80页 |
| 3.14 Masson's trichrome staining | 第80页 |
| 3.15 Statistics | 第80-81页 |
| 4. Results | 第81-94页 |
| 4.1 Mice over-expressing Handl and Twist1 mutants in cardiomyocytes developed pathological cardiac remodeling | 第81-88页 |
| 4.2 Hand1 and Twist1 could be phosphorylated by Akt in vitro and in vivo | 第88-91页 |
| 4.3 Akt suppressed Hand1 activation of reporter gene transcription | 第91-93页 |
| 4.4 Phosphorylation reduced Hand1 DNA binding ability | 第93-94页 |
| 5. Discussion | 第94-96页 |
| 6. Acknowledgements | 第96-97页 |
| 7. References | 第97-101页 |
| 致谢 | 第101-102页 |
| Publications | 第102-104页 |
