1. Oxidative Stress and Diabetes Jian Li Beijing Institute of Geriatrics Ministry of Health

2. Redox "rheostat“ in vascular cells

3. Reactive oxygen species (= ROS) O2O2 O2-O2- H 2 O 2 acidic pH, Superoxyde Dismutase (SOD) NADPH oxidase Superoxide anion Hydrogen peroxide

4. Proposed functions of ROS killing of microorganisms DNA damage cancerogenesis ageing cell death NO inactivation and peroxynitrite generation regulation of cell growth and differentiation regulation of cell function oxygen sensing activation of redox-sensitive transcription factors activation of redox-sensitive second messenger systems

5. Where and why are reactive oxygen species generated? Mitochondria – by-product of the oxidative metabolism Phagocyte NADPH oxidase – microbial killing NADPH oxidase of non-phagocytic cells –cell growth, cell signaling

6. NOX-type NADPH oxidases as superoxide-producing enzymes Fe outside inside IVIIVIIIIIV NH 2 HH H H H H NADPH FAD COOH H115 O2O2 O2-O2- e-e-

7. The NOX family of NADPH oxidases Review: Lambeth et al. Novel homologs of gp91phox.Trends in Biochemical Sciences 25: 459-461, 2000. gp91phox homology EF-hands Nox1colon Nox2phagocytes Nox3inner ear Nox4kidney Nox5 testis and lymphoid tissues O2O2 O2-O2- NADPH e-e-

8. Structure of the NAD(P)H oxidase

9. Characteristics of neutrophil and vascular NAD(P)H oxidase

10. NAD(P)H Oxidase Activation

11. Adenovirus-induced overexpression of PKC- β 2 causes the membranous translocation of p47phox and p67phox

12. A model illustrating how increased ROS production in accumulated fat contributes to metabolic syndrome

13. Mechanism for increased ROS production induced by diabetes and insulin-resistant state

14. Linking various risk factors to ROS generation in the development of IDDM

15. Initiation and amplification of the immune/inflammatory response by ROS-induced NFκ B activation in β-cell death

16. Schematic illustration of ROS-mediated NFκB activation

17. Elevated glucose and FFA levels contribute to the pathophysiology of diabetes via the generation of ROS

18. The role of serine kinase activation in oxidative stress induced insulin resistance

19. Vascular effects of reactive oxygen species (ROS)

20. Modulation of cellular function by ROS in cardiovascular diseases

21. Potential role of NADPH oxidase in the pathogenesis of diabetic nephropathy

22. Effect of high glucose level and PMA on ROS production in aortic smooth muscle cells (A) and endothelial cells (B)

23. Effect of diphenylene iodonium on high glucose– or PMA-induced increase in ROS production in aortic smooth muscle cells (A) or endothelial cells (B)

24. PKC- β inhibition suppresses diabetes-induced O 2 - production

25. Redox-dependent signaling pathways by Ang II in vascular smooth muscle cells

26. Detection of intracellular production of reactive oxygen species. A. Fluorescence microscopy visualization of ROS production in pericytes and smooth muscle cells. a: control;b: cells cultured in 25 mM glucose and AGE-Lys stimulated with Ang II;c and d : corresponding phase contrast microscopy. B. Pericytes cultured in the pro- diabetic environment, were loaded for 30 min at 37oC with 5mM DCF-DA.

27. The effect of high glucose concentration, AGE-Lys and their combination with Ang II on intracellular calcium [Ca2+]i

28. Detection of O 2 - production by dihydroethidium staining in mesangial cells overexpressing PKC- β 2

29. Superoxide production in nonatherosclerotic and atherosclerotic arteries

30. nonatherosclerotic arteries atherosclerotic arteries Expression of NAD(P)H oxidase subunits in nonatherosclerotic and atherosclerotic arteries

31. In situ detection of superoxide in sham- operated and injured carotid arteries

32. Possible antioxidative agents for diabetic vascular complications

33. Thank you!