Dissertation > Excellent graduate degree dissertation topics show

Factor 1 (TIMP - 1 ) , AG3340 islet function and rAAV-TIMP-1, LV-TIMP-1 build and identification of tissue inhibitor of metalloproteinase

Author: ZhuJian
Tutor: LiuChao;HanXiao
School: Nanjing Medical University
Course: Internal Medicine
Keywords: islet function/insμlin secreation tissue inhibitor of metalloproteinase-1/AG3340 adeno-associated virus/lentivirus
CLC: R587.1
Type: Master's thesis
Year: 2005
Downloads: 55
Quote: 0
Read: Download Dissertation

Abstract


Objective(1)To evaluate the effect of tissue inhibitor of metalloproteinase-1 (TIMP-1) and AG3340 on the insμlin secretion in interleukin-1β treatedpancreatic islets.(2)To construct and identify recombinant adeno-associated virusrAAV-TIMP-1 and lentivirus vector LV-TIMP-1 which encoding fμlllength cDNA of TIMP-1.Methods(1)Rat islets were pretreated with TIMP-1 or AG3340 1h beforeadditional IL-1β were treated and maintained 24 hours at 37℃, theglusose stimulated insulin secretion(GSIS)test were performed and thenthe medium was collected for insulin measurement.(2)Using the technique of gene recombination, the main plasmid ofpAAV-TIMP-1 was constructed. Recombinant AAV-TIMP-1 wasobtained from HEK293 cells, which were cotransfected with AAV vectorplasmids by calcium phosphate precipitation method. The titer ofrecombinant AAV was determined by counting positive HT1080 cellswhich were transduced with rAAV-GFP, finally, Western blots wereperformed to confirm the expression of rAAV-TIMP-1 in HT1080 cells.(3)Using the technique of gene recombination, the main plasmid ofpLenti6/V5-TIMP-1 was constructed. Viral supernatant was obtainedfrom 293FT cells, which were cotransfected with plasmids of lentivirusexpression system by Lipofectamine precipitation method. The titers ofthe lentiviral vector were determined by scoring GFP positive RIN5Fcells following serial dilutions of the viral supernatant transduction,finally, Western blots were performed to confirm the expression ofLV-TIMP-1 in 293FT cells.Results(1)Insμlin secreation was decreased by interleukin-1β . Islets treated withTIMP-1 or AG3340 did not show any alteration in GSIS compared withcontrol group. TIMP-1,but not AG3340 was able to restore GSIS ininterleukin-1β treated islets.(2)The titer of rAAV viral stock could be reached up to 10~9 TU/mL.Western blot results showed that rAAV-TIMP-1 expressed TIMP-1proteins in HT1080 cells.(3)The titer of lentivirus supernatant could be reached up to 10~4TU/mL.Western blot showed recombinant Lentivirus expressed TIMP-1 proteinsin 293FT cells.Conclusion(l)Tissue inhibitor of metalloproteinase-1, not AG3340, could preventinterleukin-1β mediated dysfunction in pancreatic Islets.(2)The development of recombinant rAAV-TIMP-1 and LV-TIMP-1provided a preliminary tool for TIMP-1 gene therapy in islets.

Related Dissertations

  1. Exoression of Resistin During Porcine Preadipocyte Differentiation and Its Infiuencing Factors,R587.1
  2. Serum A-FABP Levels and Its Expression in Atherosclerotic Plaques of Diabetes Mellitus,R587.1
  3. Effect of High Glucose on Chemokine Ligand 16 (CXCL16) Expression and Secretion in THP-1 Macrophages and Intervention of Aspirin in the Activity,R587.2
  4. The Effects of Atorvastatin on Expression of Osteopontin in Kidney of the Diabetic Rats Its Molecular Mechanism,R587.2
  5. The Effect and Mechanism of p38MAPK Inhibitor CBS3830 on Intimal Hyperplasia in Autogenous Vein Grafts of Diabetic Rats,R587.1
  6. Effect of β-Casomorphin7 or Casein Peptides on Blood Glucose and Oxidative Stress of the Diabetic Rats,R587.1
  7. Effects of Angiopoietin-1 on Vascular Angiogenesis Regulator Factor in Human Umbilical Vein Endothelial Cells Cultured with High Glucose,R587.2
  8. Experimental Study of Influence of Atrovastatin on Expression of NF-κB in Retionpathy of Diabetic Rats,R587.1
  9. Relationship between Level of Serum Uric Acid and Lower Extremity Arterial Disease in Patients with Type 2 Diabetes Mellitus,R587.1
  10. Low Dose of Spironolactone Reduces Oxidative Stress and Prevents the Development of Diabetic Nephropathy in Patients with Type 2 Diabetes Mellitus,R587.2
  11. Improvement of Interleukin Profiles in Patients with Type 2 Diabetes Mellitus After Add-on Acarbose Therapy Independent of Glycemic Control,R587.1
  12. Efficacy and Safety of Biphasic Insulin Aspart 30 Once-daily Treatment in Patients with Type 2 Diabetes,R587.1
  13. The Blood Glucose Control and Influencing Factors in Type 2 Diabetic Patients with Oral Hypoglycemic Agents,R587.1
  14. The Analysis of Related Factors in Type 2 Diabetes with Depression,R587.1
  15. Effect of β-Casomorphin-5 on Blood Glucose and Oxidative Stress of Diabetic Rats,R587.1
  16. The Effects of Aerobic Exercise Training on Oxidative Stress and Ultrastructure of Pancreatic Organization in Diabetic Rats,R587.1
  17. Association of Genetic Variation of UTS2 Gene with Type2 Diabetes Mellitus and Hypertension,R587.1
  18. Risk Factors of Early Onset Type 2 Diabetes with Non-alcoholic Fatty Liver Disease,R587.1
  19. Preliminary Study on Angiogenesis in Ischemic Hindlimb of Diabetic Rat Model after Mesenchymal Stem Cell Implantation,R587.1
  20. The Expression of Visfatin in Liver of Rats in Different Glucose Metabolic Statuses and Effects to Glucose and Energy Metabolism,R587.1
  21. Expression and Role of Uncoupling Protein 2 in the Kidney of Mouse after Ischemia Reperfusion,R587.1

CLC: > Medicine, health > Internal Medicine > Endocrine diseases and metabolic diseases > Islet disease > Diabetes
© 2012 www.DissertationTopic.Net  Mobile