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Research article

Effect of inhibition of glutathione synthesis on insulin action: in vivo and in vitro studies using buthionine sulfoximine

Mogher KHAMAISI, Oren KAVEL, Moti ROSENSTOCK, Michal PORAT, Michal YULI, Nurit KAISER, Assaf RUDICH
Biochemical Journal Jul 15, 2000, 349 (2) 579-586; DOI: 10.1042/bj3490579
Mogher KHAMAISI
Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 Israel
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Oren KAVEL
Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 Israel
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Moti ROSENSTOCK
The S. Daniel Abraham Center for Health and Nutrition, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 Israel
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Michal PORAT
Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 Israel
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Michal YULI
Department of Endocrinology and Metabolism, P. O. Box 12000, Hebrew University, Hadassah Medical Center, Jerusalem, 91120 Israel
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Nurit KAISER
Department of Endocrinology and Metabolism, P. O. Box 12000, Hebrew University, Hadassah Medical Center, Jerusalem, 91120 Israel
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Assaf RUDICH
Department of Clinical Biochemistry, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 IsraelThe S. Daniel Abraham Center for Health and Nutrition, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, IL-84105 Israel
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Abstract

Decreased cellular GSH content is a common finding in experimental and human diabetes, in which increased oxidative stress appears to occur. Oxidative stress has been suggested to play a causative role in the development of impaired insulin action on adipose tissue and skeletal muscle. In this study we undertook to investigate the potential of GSH depletion to induce insulin resistance, by utilizing the GSH synthesis inhibitor, L-buthionine-[S,R]-sulfoximine (BSO). GSH depletion (20-80% in various tissues), was achieved in vivo by treating rats for 20 days with BSO, and in vitro (80%) by treating 3T3-L1 adipocytes with BSO for 18 h. No demonstrable change in the GSH/GSSG ratio was observed following BSO treatment. GSH depletion was progressively associated with abnormal glucose tolerance test, which could not be attributed to impaired insulin secretion. Skeletal muscle insulin responsiveness was unaffected by GSH depletion, based on normal glucose response to exogenous insulin, 2-deoxyglucose uptake measurements in isolated soleus muscle, and on normal skeletal muscle expression of GLUT4 protein. Adipocyte insulin responsiveness in vitro was assessed in 3T3-L1 adipocytes, which displayed decreased insulin-stimulated tyrosine phosphorylation of insulin-receptor-substrate proteins and of the insulin receptor, but exaggerated protein kinase B phosphorylation. However, insulin-stimulated glucose uptake was unaffected by GSH depletion. In accordance, normal adipose tissue insulin sensitivity was observed in BSO-treated rats in vivo, as demonstrated by normal inhibition of circulating non-esterified fatty acid levels by endogenous insulin secretion. In conclusion, GSH depletion by BSO results in impaired glucose tolerance, but preserved adipocyte and skeletal muscle insulin responsiveness. This suggests that alternative oxidation-borne factors mediate the induction of peripheral insulin resistance by oxidative stress.

  • 3T3-L1 adipocytes
  • adipose tissue
  • insulin resistance
  • oxidative stress
  • skeletal muscle
  • The Biochemical Society, London © 2000
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July 2000

Volume: 349 Issue: 2

Biochemical Journal: 349 (2)
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Effect of inhibition of glutathione synthesis on insulin action: in vivo and in vitro studies using buthionine sulfoximine
Mogher KHAMAISI, Oren KAVEL, Moti ROSENSTOCK, Michal PORAT, Michal YULI, Nurit KAISER, Assaf RUDICH
Biochemical Journal Jul 2000, 349 (2) 579-586; DOI: 10.1042/bj3490579
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Effect of inhibition of glutathione synthesis on insulin action: in vivo and in vitro studies using buthionine sulfoximine
Mogher KHAMAISI, Oren KAVEL, Moti ROSENSTOCK, Michal PORAT, Michal YULI, Nurit KAISER, Assaf RUDICH
Biochemical Journal Jul 2000, 349 (2) 579-586; DOI: 10.1042/bj3490579

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Keywords

3T3-L1 adipocytes
adipose tissue
insulin resistance
oxidative stress
skeletal muscle

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