Окислительный стресс и методы терапии: как свободные радикалы провоцируют болезни и как их нейтрализуют

When there are more free radicals present than can be kept in balance by antioxidants, the free radicals can start doing damage to fatty tissue, DNA, and proteins in your body. When there are more free radicals present than can be kept in balance by antioxidants, the free radicals can start doing damage to fatty tissue, DNA, and proteins in your body. Proteins, lipids, and DNA make up a large part of your body, so that damage can lead to a vast number of diseases over time. This imbalance is associated with a number of health related concerns including heart disease, diabetes, atherosclerosis and high blood pressure. We produce a small amount of free radicals naturally, when we exercise or have inflammation, such as an impacted tooth. Oxidative stress is how we describe that imbalance between the free radicals (unstable molecules) and antioxidants in the body. This imbalance causes free radicals to damage cells, proteins and deoxyribonucleic acid (DNA), our genetic code. While the body produces free radicals during normal metabolic activity, an excess of these oxygen-containing molecules with their uneven number of electrons can cause large chemical change reactions or oxidation. Antioxidants are molecules that can donate an electron to a free radical stabilizing them without making themselves unstable. When there are more free radicals in our bodies than antioxidants, damage occurs. We see this in conditions such as diabetes, atherosclerosis (hardening of the blood vessels), inflammation, high blood pressure, heart disease, neurodegenerative diseases and cancer. Oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus and cardiovascular diseases including hypertension. The low levels of antioxidants accompanied by elevated levels of free radicals play a major role in delayed wound healing. Ultra-low microcurrent may have an antioxidant effect and has been shown to accelerate wound healing. In a study looking at ultra-low microcurrent in the management of diabetes mellitus, hypertension and chronic wounds, twelve cases were discussed with an eye to determine the mechanism of action. The purpose of the study was to investigate the efficacy of ultra-low microcurrent delivered by the Electro Pressure Regeneration Therapy (EPRT) device (EPRT Technologies-USA, Simi Valley, CA) in the management of diabetes, hypertension and chronic wounds. The EPRT device is an electrical device that sends a pulsating stream of electrons in a relatively low concentration throughout the body. The device is noninvasive and delivers electrical currents that mimic the endogenous electric energy of the human body. It is a rechargeable battery-operated device that delivers a direct current (maximum of 3 milliAmperes) of one polarity for 11.5 minutes, which then switched to the opposite polarity for another 11.5 minutes.  The resulting cycle time is approximately 23min or 0.000732 Hz and delivers a square wave bipolar current with a voltage ranging from 5V up to a maximum of 40 V. The device produces a current range of 3 mA down to 100 nA. Twelve patients with long standing diabetes, hypertension and unhealed wounds were treated with EPRT.  The patients were treated approximately for 3.5 h/day/5 days a week. Assessment of ulcer was based on scale used by National Pressure Ulcer Advisory Panel Consensus Development Conference. Patients were followed-up with daily measurement of blood pressure and blood glucose level, and their requirement for medications was recorded. Treatment continued from 2-4 months according to their response. Results showed that diabetes mellitus and hypertension were well controlled after using this device, and their wounds were markedly healed (30-100%). The patients either reduced their medication or completely stopped after the course of treatment. No side effects were reported. The mechanism of action was discussed. Diabetes mellitus and cardiovascular diseases are challenging medical and social problems. Patients with diabetes mellitus are at a higher risk of developing vascular dysfunction and hypertension. The real etiology of these diseases is not well understood. However, cumulative evidence suggests that oxidative stress may play a key role in the development of diseases. It has been found that oxidative stress is associated with several cardiovascular diseases, including atherosclerosis, hypertension, heart failure, stroke, and diabetes, and plays a fundamental role in endothelial dysfunction associated with these diseases 1 - 6. Further, an excess of free radicals plays a major role in the pathogenesis of both types of diabetes mellitus. High levels of free radicals and the decline of antioxidant defense mechanisms lead to damage of cellular organelles and enzymes, increased lipid peroxidation, and development of insulin resistance 7. The vascular and systemic complications in diabetes are associated with hyperglycemia-induced overproduction of reactive oxygen species 8, 9. Other studies showed that overproduction of reactive oxygen and nitrogen species, lowered antioxidant defense and alterations of enzymatic pathways in humans with poorly controlled diabetes mellitus can contribute to endothelial, vascular and neurovascular dysfunction 10. Insulin resistance is associated with reduced intracellular antioxidant defense, and therefore diabetic patients may have a defective intracellular antioxidant response that causes diabetic complications 11 - 13. The combination of the low levels of antioxidants and raised levels of free radical play a major role in delaying wound healing in aged rate and diabetic rats 14. It has been found that chronic leg ulcers contain localized oxidative stress 15. The recent finding revealed that insulin resistance is associated in humans with reduced intracellular antioxidant 11. Interestingly, antioxidants improve insulin sensitivity and help in wound healing 16, 17. Along with others, the investigators have used microcurrent for treatment of chronic wounds and ulcers 18 - 20. In an earlier work, The Electro Pressure Regeneration Therapy (EPRT) device which produces a current range of 3 mA down to 100 nA, was used for treatment of chronic wounds and ulcers associated with chronic disease 21.  The device used in the experiment was supposed to deliver electrons to tissues and then saturated free radicals with required electrons. The actual tissue regeneration, along with concomitant improvement noted in the general condition of the patient, points to a highly potent antioxidant effect on local tissues, as well as on tissues in general.  This reduces free radicals and might facilitate tissue repair. This device is used as a model to deliver electrons to the body, including mitochondria and presumably working as an antioxidatant device. It was thought reasonable to use on patients with diabetes mellitus, hypertension and chronic wounds, to test whether delivering electrons to the body might help eliminate underlying oxidative stress, stabilize mitochondria and prevent further formation of excess free radicals. The EPRT device is an electrical device that sends a pulsating stream of electrons in a relatively low concentration throughout the body. The device is noninvasive and delivers electrical currents that mimic the endogenous electric energy of the human body. It is a rechargeable battery-operated device that delivers a direct current (maximum of 3 milliAmperes) of one polarity for 11.5 minutes, which then switched to the opposite polarity for another 11.5 minutes. The device was designed to switch the direction of current flow halfway through the cycle. The resulting cycle time is approximately 23min or 0.000732 Hz and delivers a square wave bipolar current with a voltage ranging from 5V up to a maximum of 40 V. The device produces a current range of 3 mA down to 100 nA. Electrodes are applied in 2 layers, and tap water is used as the conducting medium. The wraps cover a large surface area, thus reducing resistance and allowing an optimum number of electrons to flow freely into tissues. There are several of these EPRT devices coming to market and one in use at the Cleveland Clinic in Ohio. Tech giants Alphabet, Amazon and Apple are all exploring new technologies in the area of diabetes care. A Canadian company, CellMedX, have recently completed clinical studies on a new microcurrent device, the eBalance, looking at these same medical markers to positive effect. The results of these preliminary trials shows that ultra-low microcurrent has therapeutic effects in the treatment of diabetes, hypertension and wound healing. Presumably, one of mechanisms of action is its antioxidant activity. The action of EPRT is to produce electrical pressure rather than an electrical jolt as produced by a Transcutaneous Electrical Nerve Stimulator. Whereas Transcutaneous Electrical Nerve Stimulator device can produce a current varying from 1uA to 100 mA, the EPRT ranges from 100 nA to 3 mA. Moreover, Transcutaneous Electrical Nerve Stimulator frequency range is from 0.5 to 40,000 Hz with a range of cycle times from 2 seconds to 0.025 milliseconds. The EPRT has a frequency of approximately 0.000732Hz which gives a frequency time of 22.77 minutes. Namely, Transcutaneous Electrical Nerve Stimulator with power of 10 mA and a frequency of 1 Hz is delivering approximately 6x10 (14) electrons per cycle. As the cycle is 1 second all these electrons were delivered in that period as a jolt. The EPRT at a setting of 100 nA is delivering 8.129x10 (14) per cycle. But as this amount is being delivered over a 23 minute period (at rate of 6x10 (11) electrons per second) this behaves as a pressure instead of a jolt. This steady stream of electrons is what makes the EPRT a super antioxidant and not only does this correct malalignments in the cells electrical system but it also eliminates free radicals and then stimulates the mitochondria to produce ATP. Microcurrent has been successfully used to enhance soft tissue healing and to treat fracture nonunion 22, 23. Microcurrent relieves myocontracture and can enhance conventional rehabilitation programs for children with cerebral palsy 24. Studies from the 1980s suggest that microcurrent therapy is effective at relieving the side effects of radiation therapy 25. The investigators have found that direct electrical therapy was effective in healing gum abscess and accelerated wound healing 20. Substances that increase electrical field, such as prostaglandin E 2, enhance the wound healing rate and increase cell division 26 - 28. Electrical fields stimulate secretion of growth factor 28. Low mA current stimulates adenosine triphosphate production 26. It is discovered in another study that microcurrent stimulates dermal fibroblasts and U937 cells to secrete transforming growth factor-β 1, a major regulator of cell-mediated inflammation and tissue regeneration 29. Insulin resistance plays a major role in the development of several metabolic abnormalities and diseases such as type 2 diabetes mellitus, obesity and the metabolic syndrome 30. In these conditions there is an elevation of both glucose and free fatty acid levels in the blood and an increase in oxidative stress 30, 31. The high degree of oxidative stress might have an important role in decreasing insulin responsiveness 31 - 33. Many studies have suggested that ß-cell dysfunction results from prolonged exposure to high glucose and elevated free fatty levels 33. High glucose concentrations induce mitochondrial reactive oxygen species, which suppresses the first phase of glucose-induced insulin secretion 34. ß-cells are particularly sensitive to reactive oxygen species because they are low in antioxidant enzymes such as catalase, glutathione peroxidase, and superoxide dismutase 35. Therefore, the oxidative stress might damage mitochondria and markedly blunt insulin secretion 34. Recent studies suggested that ß-cell lipotoxicity is enhanced by concurrent hyperglycemia and that oxidative stress may be the mediator 36, 37.  An increase in insulin, free fatty acid, and/or glucose levels can increase reactive oxygen species production and oxidative stress, as well as activate stress-sensitive pathways 33. Many studies show that postprandial hyperglycemia is associated with oxidative stress generation 38. Repeated exposure to hyperglycemia and increased levels of free fatty acid can lead to ß-cell dysfunction that may become irreversible over time. It has been suggested that oxidative stress might be the mediator of damage to cellular components of insulin production 33, 39. A major source of cellular reactive oxygen species is mitochondria, whose dysfunction contributes to pathological conditions such as vascular complications of diabetes, neurodegenerative diseases and cellular senescence 40 - 45. Source of reactive oxygen species in insulin secreting pancreatic β-cells and cells that are targets for insulin action is considered to be the mitochondrial electron transport chain. Hyperglycemia and lipotoxicity in obesity and related disorders are associated with mitochondrial dysfunction and oxidative stress 46, 47. Oxidative stress-induced activation of NF-κB signaling might be associated with the pathogenesis of insulin resistance and type 2 diabetes 48 - 51. In obesity and type 2 diabetes it has been reported that antioxidants and IKK-B inhibitors protect against insulin resistance 52, 53. The data shows that increased lipid peroxidation in NIDDM has implications for vascular disease in diabetes 54. Oxidative stress plays an important role in the pathogenesis of cardiovascular diseases including hypertension 55. Clinical studies suggest the occurrence of increased reactive oxygen species production in humans with essential hypertension 56, 57. Oxidative stress is considered to be a unifying mechanism for hypertension and atherosclerosis 58, 59. Oxygen free radicals play a major role in the failure of ischemic wound healing, while antioxidants partly improve the healing in ischemic skin wounds 60. Oxygen free radicals mediate the inhibition of wound healing following ischemia-reperfusion and sepsis 61. It seems that diabetes mellitus, cardiovascular disease, such as hypertension, and delayed wound healing have a common important basic pathogenesis, which is related to imbalance between free radical production and removal. The use of ultra-low microcurrent might help in stabilizing mitochondria, working as antioxidants and therefore, enhancing normal function of β-cells and vascular tissue. Several clinical trials have demonstrated that treatment with vitamin E, vitamin C, or glutathione improves insulin sensitivity in insulin-resistant individuals 16, 62. The acute effects of hyperglycemia-dependent endothelial cells dysfunction are counterbalanced by antioxidants 63 - 65. But clinical trials with antioxidants, in particular with vitamin E, have failed to show any beneficial effect 66. However, antioxidant therapy with vitamin E or other antioxidants is limited to scavenging already formed oxidants and may be considered symptomatic instead of a causal treatment for oxidative stress 67. Interruption of the overproduction of superoxide by the mitochondrial electron transport chain would normalize the pathways involved in the development of the oxidative stress 68. Ultra-low microcurrent therapy may change the concept of management of chronic disease. Conclusively, oxidative stress and oxidative damage to tissues are common pathology of chronic diseases, and using antioxidants in the form of ultra-low microcurrents may change the concept of how we manage chronic diseases. References: 1. Griendling KK, Fitzgerald GA. Oxidative stress and cardiovascular injury. Animal and human studies. Circulation. 2003;108:2034–2040. [ PubMed ] [ Google Scholar ] 2. Madamanchi NR, Vendrov A, Runge MS. Oxidative stress and vascular disease. Arterioscler Thromb Vasc Biol. 2005;25:29–38. [ PubMed ] [ Google Scholar ] 3. Mueller CFH, Laude K, McNally JS, Harrison DG. Redox mechanisms in blood vessels. Arterioscler Thromb Vasc Biol. 2005;25:274–278. [ PubMed ] [ Google Scholar ] 4. Steinberg D. Low density lipoprotein oxidation and its pathobiological significance. J Biol Chem. 1997;272:20963–20966. [ PubMed ] [ Google Scholar ] 5. Wei EP, Kontos HA, Christman CW, DeWitt DS. Superoxide generation and reversal of acetylcholine-induced cerebral arteriolar dilation after acute hypertension. Circ Res. 1985;57:781–787. [ PubMed ] [ Google Scholar ] 6. Rubanyi GM, Vanhoutte PM. Superoxide anions and hyperoxia inactivate endothelium-derived relaxing factor. Am J Physiol: Heart Circ Physiol. 1986;250:H822–H827. [ PubMed ] [ Google Scholar ] 7. Maritim C, Sanders R, Watkins J. Diabetes, oxidative stress, and antioxidants: A review. J Bioch Mol Toxicol. 2003;17:24– 38. [ PubMed ] [ Google Scholar ] 8. Baynes J, Thorpe S. Role of oxidative stress in diabetic complications: a new perspective on an old paradigm. Diabetes. 1999;48:1–9. [ PubMed ] [ Google Scholar ] 9. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001;414:813–820. [ PubMed ] [ Google Scholar ] 10. Jakus V. The role of free radicals, oxidative stress and antioxidant systems in diabetic vascular disease. Bratisl Lek Listy. 2000;101:541–51. [ PubMed ] [ Google Scholar ] 11. Bruce CR, Carey AL, Hawley JA, Febbraio MA. Intramuscular heat shock protein 72 and heme oxygenase-1 mRNA are reduced in patients with type 2 diabetes: evidence that insulin resistance is associated with a disturbed antioxidant defense mechanism. Diabetes. 2003;52:2338–2345. [ PubMed ] [ Google Scholar ] 12. Ceriello A, Morocutti A, Mercuri F, Quagliaro L, Moro M, Damante G, Viberti GC. Defective intracellular antioxidant enzyme production in type 1 diabetic patients with nephropathy. Diabetes. 2000;49:2170–2177. [ PubMed ] [ Google Scholar ] 13. Hodgkinson AD, Bartlett T, Oates PJ, Millward BA, Demaine AG. The response of antioxidant genes to hyperglycemia is abnormal in patients with type 1 diabetes and diabetic nephropathy. Diabetes. 2003;52:846–851. [ PubMed ] [ Google Scholar ] 14. Anamika M, Rasik AS. Antioxidant status in delayed healing type of wounds. Inter J Exper Path. 2000;81:257–263. [ PMC free article ] [ PubMed ] [ Google Scholar ] 15. Tim J, James MS, Margaret A. Hughes, George W. Cherry, Richard P. Taylor. Evidence of oxidative stress in chronic venous ulcers. Wound Rep Reg. 1999;11:172–176. [ PubMed ] [ Google Scholar ] 16. Paolisso G, Giugliano D. Oxidative stress and insulin action. Is there a relationship? Diabetologia. 1996;39:357–363. [ PubMed ] [ Google Scholar ] 17. Sen CK, Khanna S, Gordillo G, Bagchi D, Bagchi M, Roy S. Oxygen, oxidants, and antioxidants in wound healing: an emerging paradigm. Ann N Y Acad Sci. 2002 May;957:239–49. [ PubMed ] [ Google Scholar ] 18. Carley P J, Wainapel S F. Electrotherapy for acceleration of wound healing: low intensity direct current. Arch Phys Med Rehabil. 1985;66:443–446. [ PubMed ] [ Google Scholar ] 19. Nessler JP, Mass DP. Direct-current stimulation of tendon healing in vitro. Clin Orthop. 1987;217:303–312. [ PubMed ] [ Google Scholar ] 20. AL-Waili N. Electrotherapy for chronic gum and periapical abscesses. J Pak Med Assoc. 1989;39:161–162. [ PubMed ] [ Google Scholar ] 21. Lee BY, Wendell K, AL-Waili N, Butler G. Ultra-low microcurrent therapy: a novel approach for treatment of chronic resistant wounds. Adv Ther. 2007;24(6):1202–9. [ PubMed ] [ Google Scholar ] 22. Bach S, Bilgrav K, Gottrup F, Jorgensen TE. The effect of electrical current on healing skin incision: an experimental study. Eur J Surg. 1991;157:171–174. [ PubMed ] [ Google Scholar ] 23. Carley P J, Wainapel S F. Electrotherapy for acceleration of wound healing: low intensity direct current. Arch Phys Med Rehabil. 1985;66:443–446. [ PubMed ] [ Google Scholar ] 24. Mäenpää H, Jaakkola R, Sandström M, Von Wendt L. Does microcurrent stimulation increase the range of movement of ankle dorsiflexion in children with cerebral palsy? Disabil Rehabil. 2004;26:669–677.[ PubMed ] [ Google Scholar ] 25. King GE, Jacob RF, Martin JW. Electrotherapy and hyperbaric oxygen: Promising treatments for postradiation complications. J Prosthetic Dentistry. 1989;62:331–334. [ PubMed ] [ Google Scholar ] 26. Cheng N, Van Hoof H, Bockx E, Hoogmartens MJ, Mulier JC, De Dijcker FJ, Sansen WM, De Loecker W. The effects of electric currents on ATP generation, protein synthesis, and membrane transport of rat skin. Clin Orthop Relat Res. 1982;171:264–72. [ PubMed ] [ Google Scholar ] 27. McCaig D, Rajnicek M, Song B, Zhao M. Has electrical growth cone guidance found its potential? Trends Neurosci. 2002;25:354–359. [ PubMed ] [ Google Scholar ] 28. Zhao M, Bai H, Wang E, Forrester V, McCaig D. Electrical stimulation directly induces pre-angiogenic response in vascular endothelial cells by signaling through VEGF receptors. J Cell Sci. 2003;117:397–405.[ PMC free article ] [ PubMed ] [ Google Scholar ] 29. Todd I, Clothier RH, Huggins ML, Patel N, Searle KC, Jeyarajah S, Pradel L, Lacey KL. Electrical stimulation of transforming growth factor-beta 1 secretion by human dermal fibroblasts and the U937 human monocytic cell line. Altern Lab Anim. 2001;29:693–701. [ PubMed ] [ Google Scholar ] 30. Petersen KF, Shulman GI. New insights into the pathogenesis of insulin resistance in humans using magnetic resonance spectroscopy. Obesity (Silver Spring) 2006;14(Suppl 1):34S–40S. [ PMC free article ][ PubMed ] [ Google Scholar ] 31. Boden G. Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes. 1997;46:3–10. [ PubMed ] [ Google Scholar ] 32. Evans JL, Maddux BA, Goldfine ID. The molecular basis for oxidative stress-induced insulin resistance. Antioxid Redox Signal. 2005;7:1040–1052. [ PubMed ] [ Google Scholar ] 33. Evans JL, Goldfine ID, Maddux BA, Grodsky GM. Are oxidative stress-activated signaling pathways mediators of insulin resistance and ß-cell dysfunction? Diabetes. 2003;52:1–8. [ PubMed ] [ Google Scholar ] 34. Robertson RP, Harmon J, Tran PO, Tanaka Y, Takahashi H. Glucose toxicity in ß-cells: type 2 diabetes, good radicals gone bad, and the glutathione connection. Diabetes. 2003;52:581–587. [ PubMed ] [ Google Scholar ] 35. Tiedge M, Lortz S, Drinkgern J, Lenzen S. Relation between antioxidant enzyme gene expression and antioxidative defense status of insulin producing cells. Diabetes. 1997;46:1733–1742. [ PubMed ] [ Google Scholar ] 36. El-Assad W, Buteau J, Peyot ML, Nolan C, Roduit R, Hardy S. et al.Saturated fatty acids synergize with elevated glucose to cause pancreatic beta-cell death. Endocrinology. 2003;144:4154–4163. [ PubMed ] [ Google Scholar ] 37. Piro S, Anello M, Di Pietro C, Lizzio MN, Patane G, Rabuazzo AM, Vigneri R, Purrello M, Purrello F. Chronic exposure to free fatty acids or high glucose induces apoptosis in rat pancreatic islets: possible role of oxidative stress. Metabolism. 2002;51:1340–1347. [ PubMed ] [ Google Scholar ] 38. Ceriello A. The possible role of postprandial hyperglycaemia in the pathogenesis of diabetic complications. Diabetologia. 2003;46:M9–M16. [ PubMed ] [ Google Scholar ] 39. Del Prato S. Loss of early insulin secretion leads to postprandial hyperglycaemia. Diabetologia. 2003;46:M2–M8. [ PubMed ] [ Google Scholar ] 40. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of ageing. Nature. 2000;408:239–247. [ PubMed ] [ Google Scholar ] 41. Huang H, Manton KG. The role of oxidative damage in mitochondria during aging: a review. Front Biosci. 2004;9:1100–111. [ PubMed ] [ Google Scholar ] 42. Nishikawa T, Edelstein D, Du XL, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates PJ, Hammes HP, Giardino I, Brownlee M. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature. 2000;404:787–790. [ PubMed ] [ Google Scholar ] 43. Jenner P. Parkinson's disease, pesticides and mitochondrial dysfunction. Trends Neurosci. 2001;24:245–247. [ PubMed ] [ Google Scholar ] 44. Aliev G, Seyidova D, Lamb BT, Obrenovich ME, Siedlak SL, Vinters HV, Friedland RP, LaManna JC, Smith MA, Perry G. Mitochondria and vascular lesions as a central target for the development of Alzheimer's disease and Alzheimer disease-like pathology in transgenic mice. Neurol Res. 2003;25:665–674. [ PubMed ] [ Google Scholar ] 45. Yorek MA. The role of oxidative stress in diabetic vascular and neural disease. Free Radic Res. 2003;37:471–480. [ PubMed ] [ Google Scholar ] 46. Schrauwen P, Hesselink MK. Oxidative capacity, lipotoxicity, and mitochondrial damage in type 2 diabetes. Diabetes. 2004;53:1412–1417. [ PubMed ] [ Google Scholar ] 47. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001;414:813–820. [ PubMed ] [ Google Scholar ] 48. Arkan MC, Hevener AL, Greten FR, Maeda S, Li ZW, Long JM, Wynshaw-Boris A, Poli G, Olefsky J, Karin M. IKK-beta links inflammation to obesity-induced insulin resistance. Nat Med. 2005;11:191–198.[ PubMed ] [ Google Scholar ] 49. Cai D, Yuan M, Frantz DF, Melendez PA, Hansen L, Lee J, Shoelson SE. Local and systemic insulin resistance resulting from hepatic activation of IKK-beta and NF-kappaB. Nat Med. 2005;11:183–190.[ PMC free article ] [ PubMed ] [ Google Scholar ] 50. Ho E, Bray TM. Antioxidants, NFkappaB activation, and diabetogenesis. Proc Soc Exp Biol Med. 1999;222:205–213. [ PubMed ] [ Google Scholar ] 51. Kim JK, Kim YJ, Fillmore JJ, Chen Y, Moore I, Lee J, Yuan M, Li ZW, Karin M, Perret P, Shoelson SE, Shulman GI. Prevention of fat-induced insulin resistance by salicylate. J Clin Invest. 2001;108:437–446. [ PMC free article ] [ PubMed ] [ Google Scholar ] 52. Evans L. Antioxidants: do they have a role in the management of insulin resistance. Indian J Med Res. 2007;125:355–375. [ PubMed ] [ Google Scholar ] 53. Honjo T, Inane N. Antioxidants drugs as the strategy for treatment of metabolic syndrome. Nippon Rinsho. 2006;28:660–667. [ PubMed ] [ Google Scholar ] 54. Davì G, Ciabottoni G, Consoli A, Messetti A, Falco A, Santarone S, Pennese E, Vitacolonna E, Bucciarelli T, Costantini F, Capani F, Patrono C. In vivo formation of 8-iso-prostaglandin F2α and platelet activation in diabetes mellitus. effects of improved metabolic control. Circulation. 1999;99:224–229.[ PubMed ] [ Google Scholar ] 55. Cai H, Harrison DG. Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res. 2000;87:840–844. [ PubMed ] [ Google Scholar ] 56. Mehta JL, Lopez LM, Chen L, Cox OE. Alterations in nitric oxide synthase activity, superoxide anion generation, and platelet aggregation in systemic hypertension, and effects of celiprolol. Am J Cardiol. 1994;74:901–905. [ PubMed ] [ Google Scholar ] 57. Lacy F, O'Connor DT, Schmid-Schonbein GW. Plasma hydrogen peroxide production in hypertensives and normotensive subjects at genetic risk of hypertension. J Hypertens. 1998;16:291–303. [ PubMed ] [ Google Scholar ] 58. Zalba G, San Jose G, Moreno M, Fortuno M, Fortuno A, Beaumont F, Diez J. Oxidative stress in arterial hypertension: role of NAD(P)H oxidase. Hypertension. 2001;38:1395–1399. [ PubMed ] [ Google Scholar ] 59. Nickenig G, Harrison DG. The AT(1)-type angiotensin receptor in oxidative stress and atherogenesis: part I: oxidative stress and atherogenesis. Circulation. 2002;105:393–396. [ PubMed ] [ Google Scholar ] 60. Senel O, Cetinkale O, Ozbay G, Ahçioğlu F, Bulan R. Oxygen free radicals impair wound healing in ischemic rat skin. Ann Plast Surg. 1997;39:516–23. [ PubMed ] [ Google Scholar ] 61. Foschi D, Trabucchi E, Musazzi M, Castoldi L, Di Mattia D, Radaelli E, Marazzi M, Franzini P, Berlusconi A. The effects of oxygen free radicals on wound healing. Int J Tissue React. 1988;10(6):373–9.[ PubMed ] [ Google Scholar ] 62. Ceriello A. Oxidative stress and glycemic regulation. Metabolism. 2000;49:27–29. [ PubMed ] [ Google Scholar ] 63. Tesfamariam B, Cohen RA. Free radicals mediate endothelial cell dysfunction caused by elevated glucose. Am J Physiol. 1992;263:H321–H326. [ PubMed ] [ Google Scholar ] 64. Marfella R, Verrazzo G, Acampora R, La Marca C, Giunta R, Lucarelli C, Paolisso G, Ceriello A, Giugliano D. Glutathione reverses systemic hemodynamic changes by acute hyperglycemia in healthy subjects. Am J Physiol. 1995;268:E1167–E1173. [ PubMed ] [ Google Scholar ] 65. Ting HH, Timimi FK, Boles KS, Creager SJ, Ganz P, Creager MA. Vitamin C improves endothelium-dependent vasodilation in patients with non-insulin-dependent diabetes mellitus. J Clin Invest. 1996;97:22–28. [ PMC free article ] [ PubMed ] [ Google Scholar ] 66. Marchioli R, Schweiger C, Levantesi G, Gavazzi L, Valagussa F. Antioxidant vitamins and prevention of cardiovascular disease: epidemiological and clinical trial data. Lipids. 2001;36:S53–S63. [ PubMed ] [ Google Scholar ] 67. Cuzzocrea S, Riley DP, Caputi AP, Salvemini D. Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury. Pharmacol Rev. 2001;53:135–159. [ PubMed ] [ Google Scholar ] 68. Ceriello A. New insights on oxidative stress and diabetic complications may lead to a "Causal" antioxidant therapy. Diabetes Care. 2003;26:1589–1596. [ PubMed ] [ Google Scholar ] 69. Lee, Bok Y et al. “Ultra-low microcurrent in the management of diabetes mellitus, hypertension and chronic wounds: report of twelve cases and discussion of mechanism of action.” International journal of medical sciences vol. 7,1 29-35. 6 Dec. 2009, doi:10.7150/ijms.7.29