| 中文摘要 | 第3-5页 |
| ABSTRACT | 第5-7页 |
| ABBREVIATION | 第15-17页 |
| 1 BACKGROUND | 第17-23页 |
| 1.1 PATHOPHYSIOLOGY OF CRITICAL LIMB ISCHEMIA | 第17-19页 |
| 1.2 ROLES OF PLATELET-DERIVED GROWTH FACTOR-BB (PDGF-BB) AND VASCULARENDOTHELIAL GROWTH FACTOR (VEGF) IN ANGIOGENESIS | 第19页 |
| 1.3 ROLE OF PROLYL HYDROXYLASE IN ANGIOGENESIS | 第19-21页 |
| 1.4 SALIDROSIDE | 第21页 |
| 1.5 STUDY PURPOSE AND INNOVATION | 第21-22页 |
| 1.6 STUDY PLAN | 第22-23页 |
| 2 EFFECT OF SALIDROSIDE ON PROMOTING ANGIOGENIC FACTORS IN SKELETAL MUSCLE CELLS IN HIGH GLUCOSECONDITION | 第23-45页 |
| 2.1 MATERIALS | 第23-27页 |
| 2.1.1 Cell isolation | 第23页 |
| 2.1.2 Equipment | 第23-24页 |
| 2.1.3 Reagents | 第24-26页 |
| 2.1.4 Primers | 第26页 |
| 2.1.5 Solution Preparation | 第26-27页 |
| 2.2 METHOD | 第27-38页 |
| 2.2.1 Cell culture | 第27-31页 |
| 2.2.2 RNA Experiment | 第31-34页 |
| 2.2.3 Protein experiment | 第34-38页 |
| 2.3 RESULTS | 第38-42页 |
| 2.3.1 Effect of salidroside on the expression levels of angiogenic factors in skeletal musclecells under high glucose condition | 第38-40页 |
| 2.3.2 Effect of salidroside on PHDs family in skeletal muscle cell under high glucosecondition | 第40-42页 |
| 2.3.3 Effect of PHD3-silencing on the expression of angiogenic factors in skeletal musclecells under high glucose condition | 第42页 |
| 2.4 DISCUSSION AND CONCLUSION | 第42-45页 |
| 3 EFFECT OF SALIDROSIDE ON SKELETAL MUSCLE CELL MOBILITY AND PROLIFERATION IN HIGH GLUCOSECONDITION | 第45-55页 |
| 3.1 MATERIALS | 第45-48页 |
| 3.1.1 Cell Isolation | 第45页 |
| 3.1.2 Equipment | 第45-46页 |
| 3.1.3 Reagents | 第46-47页 |
| 3.1.4 Solution Preparation | 第47-48页 |
| 3.2 METHOD | 第48-51页 |
| 3.2.1 Cell culture | 第48-49页 |
| 3.2.2 Transwell Chamber Assay | 第49-50页 |
| 3.2.3 Ki67 Staining | 第50-51页 |
| 3.3 RESULTS | 第51-53页 |
| 3.3.1 Effect of salidroside in skeletal muscle cell migration under high glucose condition . 49 | 第51-52页 |
| 3.3.2 Effect of salidroside in skeletal muscle cell proliferation under high glucose condition | 第52-53页 |
| 3.4 DISCUSSION AND CONCLUSION | 第53-55页 |
| 4 OVEREXPRESSION PHD3 INHIBITED THE EFFECT OFSALIDROSIDE ON PROMOTING ANGIOGENESIS IN HIGHGLUCOSE CONDITION | 第55-75页 |
| 4.1 MATERIALS | 第55-60页 |
| 4.1.1 Cell isolation | 第55页 |
| 4.1.2 Equipment | 第55-56页 |
| 4.1.3 Reagents | 第56-58页 |
| 4.1.4 Primer | 第58页 |
| 4.1.5 Solution Preparation | 第58-60页 |
| 4.2 METHOD | 第60-72页 |
| 4.2.1 Plasmid reconstruction | 第60-64页 |
| 4.2.2 Cell culture | 第64-65页 |
| 4.2.3 Transfection | 第65页 |
| 4.2.4 RNA experiment | 第65-68页 |
| 4.2.5 Protein experiment | 第68-72页 |
| 4.3 RESULTS | 第72-74页 |
| 4.3.1 Overexpression PHD3 inhibited the effect of salidroside in promoting angiogenicfactors under high glucose condition | 第72-74页 |
| 4.4 DISCUSSION AND CONCLUSION | 第74-75页 |
| 5 EFFECT OF CONDITIONED MEDIUM OF SKELETALMUSCLE CELL SECRETOME IN HUMAN ENDOTHELIAL CELLAND SMOOTH MUSCLE CELL MOBILITY, DISTRIBUTION OF FILAMENT ACTIN AND PROLIFERATION UNDER HIGHGLUCOSE CONDITION | 第75-91页 |
| 5.1 MATERIAL | 第75-78页 |
| 5.1.1 Cell isolation | 第75页 |
| 5.1.2 Equipment | 第75-76页 |
| 5.1.3 Reagent | 第76-77页 |
| 5.1.4 Solution Preparation | 第77-78页 |
| 5.2 METHOD | 第78-83页 |
| 5.2.1 Condition medium preparation | 第78页 |
| 5.2.2 Cell culture | 第78-80页 |
| 5.2.3 Transwell chamber assay | 第80页 |
| 5.2.4 Phalloidin staining | 第80-81页 |
| 5.2.5 Ki67 staining | 第81-83页 |
| 5.3 RESULTS | 第83-88页 |
| 5.3.1 Effect of conditioned medium of skeletal muscle cell in human endothelial cellmigration under high glucose condition | 第83-84页 |
| 5.3.2 Effect of conditioned medium of skeletal muscle cell in human endothelial cellfilament actin distribution under high glucose condition | 第84-85页 |
| 5.3.3 Effect of conditioned medium of skeletal muscle cell in human endothelial cellproliferation under high glucose condition | 第85页 |
| 5.3.4 Effect of conditioned medium of skeletal muscle cell in smooth muscle cell migrationunder high glucose condition | 第85-86页 |
| 5.3.5 Effect of conditioned medium of skeletal muscle cell in smooth muscle cell filamentactin distribution under high glucose condition | 第86-87页 |
| 5.3.6 Effect of conditioned medium of skeletal muscle cell in smooth muscle cellproliferation under high glucose condition | 第87-88页 |
| 5.4 DISCUSSION AND CONCLUSION | 第88-91页 |
| 6 CONDITIONED MEDIUM OF SKELETAL MUSCLE CELL THAT CONTAINING OVEREXPRESSION PHD3 INHIBITED THEEFFECT OF SALIDROSIDE ON INDUCING HUMAN ENDOTHELIAL CELL AND SMOOTH MUSCLE CELLMOBILITY AND PARACRINE FUNCTION UNDER HIGHGLUCOSE CONDITION | 第91-105页 |
| 6.1 MATERIALS | 第91-94页 |
| 6.1.1 Cell isolation | 第91页 |
| 6.1.2 Equipment | 第91-92页 |
| 6.1.3 Reagents | 第92-93页 |
| 6.1.4 Solution Preparation | 第93-94页 |
| 6.2 METHOD | 第94-97页 |
| 6.2.1 Overexpression PHD3 conditioned medium preparation | 第94页 |
| 6.2.2 Transwell chamber assay | 第94-95页 |
| 6.2.3 Phalloidin staining | 第95-96页 |
| 6.2.4 Ki67 Staining | 第96-97页 |
| 6.3 RESULTS | 第97-103页 |
| 6.3.1 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing human endothelial cell migration in high glucosecondition | 第97-98页 |
| 6.3.2 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing filament actin distribution of human endothelialcell under high glucose condition | 第98-99页 |
| 6.3.3 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing human endothelial cell proliferation under highglucose condition | 第99-100页 |
| 6.3.4 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing smooth muscle cell migration under high glucosecondition | 第100-101页 |
| 6.3.5 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing filament actin distribution of smooth muscle cellunder high glucose condition | 第101-102页 |
| 6.3.6 Conditioned medium of skeletal muscle cell that containing overexpression PHD3 inhibited the effect of salidroside on inducing smooth muscle cell proliferation under highglucose condition | 第102-103页 |
| 6.4 DISCUSSION AND CONCLUSION | 第103-105页 |
| 7 EFFECT OF SALIDROSIDE ON CRITICAL LIMB ISCHEMIAOF STREPTOZOTOCIN-INDUCED DIABETIC MICE MODEL | 第105-119页 |
| 7.1 MATERIALS | 第105-107页 |
| 7.1.1 Animal | 第105页 |
| 7.1.2 Equipment | 第105-106页 |
| 7.1.3 Reagents | 第106-107页 |
| 7.1.4 Solution preparation | 第107页 |
| 7.2 METHOD | 第107-110页 |
| 7.2.1 Streptozotocin-induced diabetes type I mice model | 第107-108页 |
| 7.2.2 Hind limb ischemia mice model | 第108页 |
| 7.2.3 Blood perfusion assessment | 第108页 |
| 7.2.4 Salidroside treatment | 第108页 |
| 7.2.5 Evaluation on hind limb ischemia | 第108页 |
| 7.2.6 Paraffin-embedded sectioning of tissue | 第108-109页 |
| 7.2.7 Immunohistochemistry staining | 第109页 |
| 7.2.8 Hematoxylin & Eosin staining | 第109-110页 |
| 7.2.9 Statistical Analysis | 第110页 |
| 7.3 RESULTS | 第110-117页 |
| 7.3.1 Effect of salidroside on blood recovery in hind limb ischemia of streptozotocin-induced diabetic mice model | 第110-112页 |
| 7.3.2 Effect of salidroside on neovascularization in hind limb ischemia of streptozotocin-induced of diabetic mice model | 第112-114页 |
| 7.3.3 Effect of salidroside on morphologies changes of the hind limb ischemia ofstreptozotocin-induced diabetic mice model | 第114-115页 |
| 7.3.4 Effect of Salidroside on angiogenic factors expression level of ischemic hind limb ofstreptozotocin-induced diabetic mice model | 第115-116页 |
| 7.3.5 Effect of salidroside on PHDs family expression level of ischemic hind limb ofstreptozotocin-induced diabetic mice model | 第116-117页 |
| 7.4 DISCUSSION AND CONCLUSION | 第117-119页 |
| 8 CONCLUSION AND FUTURE PROSPECTS | 第119-123页 |
| 8.1 CONCLUSION | 第119-122页 |
| 8.2 FUTURE PROSPECTS | 第122-123页 |
| ACKNOWLEDGEMENT | 第123-125页 |
| REFERENCES | 第125-128页 |
