1 / 15

Effect of Amifostine On Mitochondrial Superoxide Dismutase Expression In INS-1 Beta Cells

Effect of Amifostine On Mitochondrial Superoxide Dismutase Expression In INS-1 Beta Cells. Daniel Turner-Lloveras Dr. Michael Roe. Type 2 Diabetes Mellitus. Insulin Resistance Beta Cell Dysfunction Hyperglycemia Oxidative Stress (ROS) Beta Cell Degradation Beta Cell Death

aaron
Download Presentation

Effect of Amifostine On Mitochondrial Superoxide Dismutase Expression In INS-1 Beta Cells

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Effect of Amifostine On Mitochondrial Superoxide Dismutase Expression In INS-1 Beta Cells Daniel Turner-Lloveras Dr. Michael Roe

  2. Type 2 Diabetes Mellitus • Insulin Resistance • Beta Cell Dysfunction • Hyperglycemia • Oxidative Stress (ROS) • Beta Cell Degradation • Beta Cell Death • Complications of Diabetes • Blindness, Heart Disease, Kidney Problems, etc.

  3. Endogenous Antioxidant Pathways normal [glucose] H2O MnSOD Glutathione Peroxidase O2- H2O2 Catalase H2O O2

  4. Enhanced Endogenous Antioxidant Defense during hyperglycemia? HIGH [GLUCOSE] H2O H2O2 O2- MnSOD(?) Glutathione Peroxidase Catalase(?) (high mitochondrial metabolic activity) HO-- H2O O2 Oxidative Damage

  5. Hypothesis • Amifostine induces upregulation of MnSOD (Sod2) via activation of NF-κβ

  6. Amifostine • A phosphorothioate that is metabolized to the free thiol metabolite WR-1065. • Developed by military to protect soldiers during nuclear warfare. • The only drug approved by the FDA specifically as a radioprotector.

  7. Amifostine ↑ NF-κβ ↑ MnSOD HIGH [GLUCOSE] H2O H2O2 O2- MnSOD(?) Glutathione Peroxidase Catalase(?) HO-- H2O O2 Oxidative Damage

  8. METHODS • INS-1 Rat Insulinoma Cell Line • 40 µM WR-1065 • Real-Time/ Quantitative PCR (QPCR) • Western Immunoblotting 0 4 8 24 48 30 min Treatment Hours

  9. QPCR

  10. QPCR DATA • Drug induced transient increase in MnSOD gene expression • No data for Glutathione Peroxidase and Catalase

  11. Western Immunoblotting MnSOD MW 0 -- 4 + -- 8 + -- 24 + -- 48 + β-Actin -- = Control + = Treated with WR-1065

  12. Western Blot DATA • Transient Increase in MnSOD levels in both drug treated and control

  13. CONCLUSION • Amifostine upregulates sod2 (MnSOD) gene expression. • Effects on MnSOD protein levels are uncertain.

  14. Future Directions • Refine Technique and repeat experiments • Investigate role of NF-κB • Mitochondrial function and insulin secretion • In vivo experiments with Fatty Zucker Diabetic Rats

  15. Acknowledgements • Dr. Michael Roe • Dr. David Grdina • Dr. Jeff Murley • Luis Landa • Dr. Mark Harbeck • Kelly Kaihara • Dr. Kajorn Kitiphongspattana • Dr. Vernon Towle • Dr. Eugene Chang

More Related