Mitochondrial nutrients improve immune dysfunction in the type 2 diabetic Goto-Kakizaki rats

The development of type 2 diabetes is accompanied by decreased immune function and the mechanisms are unclear. We hypothesize that oxidative damage and mitochondrial dysfunction may play an important role in the immune dysfunction in diabetes. In the present study, we investigated this hypothesis in diabetic Goto‐Kakizaki rats by treatment with a combination of four mitochondrial‐targeting nutrients, namely, R‐α‐lipoic acid, acetyl‐L‐carnitine, nicotinamide and biotin. We first studied the effects of the combination of these four nutrients on immune function by examining cell proliferation in immune organs (spleen and thymus) and immunomodulating factors in the plasma. We then examined, in the plasma and thymus, oxidative damage biomarkers, including lipid peroxidation, protein oxidation, reactive oxygen species, calcium and antioxidant defence systems, mitochondrial potential and apoptosis‐inducing factors (caspase 3, p53 and p21). We found that immune dysfunction in these animals is associated with increased oxidative damage and mitochondrial dysfunction and that the nutrient treatment effectively elevated immune function, decreased oxidative damage, enhanced mitochondrial function and inhibited the elevation of apoptosis factors. These effects are comparable to, or greater than, those of the anti‐diabetic drug pioglitazone. These data suggest that a rational combination of mitochondrial‐targeting nutrients may be effective in improving immune function in type 2 diabetes through enhancement of mitochondrial function, decreased oxidative damage, and delayed cell death in the immune organs and blood.

[1]  M. Belvisi,et al.  Peroxisome proliferator-activated receptor gamma agonists as therapy for chronic airway inflammation. , 2006, European journal of pharmacology.

[2]  C. Palmeira,et al.  Alterations of liver mitochondrial bioenergetics in diabetic Goto-Kakizaki rats. , 1999, Metabolism: clinical and experimental.

[3]  G. Steele,et al.  Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[4]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[5]  P. Oliveira,et al.  Diabetes induces metabolic adaptations in rat liver mitochondria: role of coenzyme Q and cardiolipin contents. , 2003, Biochimica et biophysica acta.

[6]  C. Palmeira,et al.  Brain and liver mitochondria isolated from diabeticGoto‐Kakizaki rats show different susceptibility to induced oxidative stress , 2001, Diabetes/metabolism research and reviews.

[7]  Jiankang Liu,et al.  Stress, Aging, and Brain Oxidative Damage , 1999, Neurochemical Research.

[8]  M. Kuwajima,et al.  A role of nicotinamide-induced increase in pancreatic beta-cell mass on blood glucose control after discontinuation of the treatment in partially pancreatectomized OLETF rats. , 1998, Diabetes research and clinical practice.

[9]  M. De la Fuente,et al.  The immune system in the oxidative stress conditions of aging and hypertension: favorable effects of antioxidants and physical exercise. , 2005, Antioxidants & redox signaling.

[10]  M. Gronski,et al.  Death pathways in T cell homeostasis and their role in autoimmune diabetes. , 2006, The review of diabetic studies : RDS.

[11]  Ken-ichi Isobe,et al.  An inhibitor of mitochondrial complex I, rotenone, inactivates proteasome by oxidative modification and induces aggregation of oxidized proteins in SH‐SY5Y cells , 2003, Journal of neuroscience research.

[12]  R. Hotchkiss,et al.  Apoptosis and caspases regulate death and inflammation in sepsis , 2006, Nature Reviews Immunology.

[13]  B. Ames,et al.  (R)-alpha-lipoic acid protects retinal pigment epithelial cells from oxidative damage. , 2005, Investigative ophthalmology & visual science.

[14]  G. Mingrone.  Carnitine in Type 2 Diabetes , 2004, Annals of the New York Academy of Sciences.

[15]  Jiankang Liu.  The Effects and Mechanisms of Mitochondrial Nutrient α-Lipoic Acid on Improving Age-Associated Mitochondrial and Cognitive Dysfunction: An Overview , 2007, Neurochemical Research.

[16]  B. Ames,et al.  (R)-α-Lipoic Acid Protects Retinal Pigment Epithelial Cells from Oxidative Damage , 2005 .

[17]  F. Russo,et al.  Estrogenic regulation of cholesterol biosynthesis and cell growth in DLD-1 human colon cancer cells , 2005, Scandinavian journal of gastroenterology.

[18]  C. Palmeira,et al.  Higher efficiency of the liver phosphorylative system in diabetic Goto‐Kakizaki (GK) rats , 1999, FEBS letters.

[19]  M. Mccarty,et al.  Nutraceutical resources for diabetes prevention--an update. , 2005, Medical hypotheses.

[20]  D. Chan.  Mitochondria: Dynamic Organelles in Disease, Aging, and Development , 2006, Cell.

[21]  J. Satoh,et al.  Improvement of glucose tolerance with immunomodulators in type 2 diabetic animals , 1996, Biotherapy.

[22]  B. Ames,et al.  Reducing mitochondrial decay with mitochondrial nutrients to delay and treat cognitive dysfunction, Alzheimer's disease, and Parkinson's disease , 2005, Nutritional neuroscience.

[23]  David W. Killilea,et al.  Age-associated mitochondrial oxidative decay: Improvement of carnitine acetyltransferase substrate-binding affinity and activity in brain by feeding old rats acetyl-l- carnitine and/or R-α-lipoic acid , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[24]  A. Abbas,et al.  Cellular and Molecular Immunology , 1991 .

[25]  C. Cotman,et al.  R-α-Lipoic acid and acetyl-l-carnitine complementarily promote mitochondrial biogenesis in murine 3T3-L1 adipocytes , 2007, Diabetologia.

[26]  N. Wiernsperger,et al.  Metformin delays the manifestation of diabetes and vascular dysfunction in Goto–Kakizaki rats by reduction of mitochondrial oxidative stress , 2006, Diabetes/metabolism research and reviews.

[27]  M. S. Santos,et al.  Diabetes and mitochondrial bioenergetics: Alterations with age , 2003, Journal of biochemical and molecular toxicology.

[28]  K. Yagi,et al.  Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. , 1979, Analytical biochemistry.

[29]  E. Donadi,et al.  Effect of metabolic control on the in vitro proliferation of peripheral blood mononuclear cells in type 1 and type 2 diabetic patients. , 2006, Sao Paulo medical journal = Revista paulista de medicina.

[30]  H. Kaneto,et al.  Involvement of oxidative stress in the pathogenesis of diabetes. , 2006, Antioxidants & redox signaling.

[31]  F. Chang,et al.  Decreased cell-mediated immunity in patients with non-insulin-dependent diabetes mellitus. , 1995, Diabetes research and clinical practice.

[32]  B. Ames,et al.  Acetyl‐L‐carnitine and α‐lipoic acid supplementation of aged beagle dogs improves learning in two landmark discrimination tests , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[33]  James C Bartholomew,et al.  Feeding acetyl-l-carnitine and lipoic acid to old rats significantly improves metabolic function while decreasing oxidative stress , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[34]  B. Portha.  Programmed disorders of β‐cell development and function as one cause for type 2 diabetes? The GK rat paradigm , 2005, Diabetes/metabolism research and reviews.

[35]  S. Khandelwal,et al.  Role of oxidative stress and apoptosis in cadmium induced thymic atrophy and splenomegaly in mice. , 2007, Toxicology letters.

[36]  K. Maiese,et al.  Oxidative stress biology and cell injury during type 1 and type 2 diabetes mellitus. , 2007, Current neurovascular research.

[37]  B. Ames,et al.  Oxidative damage and mitochondrial decay in aging. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[38]  A. De Gaetano,et al.  Acetyl-L-carnitine infusion increases glucose disposal in type 2 diabetic patients. , 2000, Metabolism: clinical and experimental.

[39]  M. Hanefeld,et al.  Treatment of symptomatic diabetic peripheral neuropathy with the anti-oxidant alpha-lipoic acid. A 3-week multicentre randomized controlled trial (ALADIN Study). , 1995, Diabetologia.

[40]  S. Riley,et al.  The quest for a model of type II diabetes with nephropathy: the Goto Kakizaki rat. , 2004, Journal of nephrology.

[41]  S. Yamamoto,et al.  Exercise training restores decreased cellular immune functions in obese Zucker rats. , 1998, Journal of applied physiology.

[42]  W. Habig,et al.  Glutathione S-transferase A. A novel kinetic mechanism in which the major reaction pathway depends on substrate concentration. , 1974, The Journal of biological chemistry.

[43]  J. Fanzo,et al.  p53 protein and p21 mRNA levels and caspase-3 activity are altered by zinc status in aortic endothelial cells. , 2002, American journal of physiology. Cell physiology.

[44]  L. Packer,et al.  Cytosolic and Mitochondrial Systems for NADH- and NADPH-dependent Reduction of α-Lipoic Acid , 1997 .

[45]  M. Kirsch,et al.  NAD(P)H, a directly operating antioxidant? , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[46]  E. Stadtman,et al.  Carbonyl assays for determination of oxidatively modified proteins. , 1994, Methods in enzymology.

[47]  C. Östenson,et al.  Diabetic type II Goto-Kakizaki rats show progressively decreasing exploratory activity and learning impairments in fixed and progressive ratios of a lever-press task , 2007, Behavioural Brain Research.

[48]  B. Ames,et al.  Memory loss in old rats is associated with brain mitochondrial decay and RNA/DNA oxidation: Partial reversal by feeding acetyl-l-carnitine and/or R-α-lipoic acid , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[49]  R. Mccaman,et al.  Carnitine acetyltransferase in nervous tissue. , 1966, The Journal of biological chemistry.

[50]  J. Hayes,et al.  Glutathione S‐transferases , 2002 .