Acceleration of protein glycation by oxidative stress and comparative role of antioxidant and protein glycation inhibitor
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Hyperglycemia in diabetes causes protein glycation that leads to oxidative stress, release of cytokines, and establishment of secondary complications such as neuropathy, retinopathy, and nephropathy. Several other metabolic disorders, stress, and inflammation generate free radicals and oxidative stress. It is essential to study whether oxidative stress independently enhances protein glycation leading to rapid establishment of secondary complications. Oxidative stress was experimentally induced using rotenone and Fenton reagent for in vivo and in vitro studies, respectively. Results showed significant increase in the rate of modification of BSA in the form of fructosamine and protein-bound carbonyls in the presence of fenton reagent. Circular dichroism studies revealed gross structural changes in the reduction of alpha helix structure and decreased protein surface charge was confirmed by zeta potential studies. Use of rotenone demonstrated enhanced AGE formation, ROS generation, and liver and kidney tissue glycation through fluorescence measurement. Similar findings were also observed in cell culture studies. Use of aminoguanidine, a protein glycation inhibitor, demonstrated reduction in these changes; however, a combination of aminoguanidine along with vitamin E demonstrated better amelioration. Thus, oxidative stress accelerates the process of protein glycation causing gross structural changes and tissue glycation in insulin-independent tissues. Use of antioxidants and protein glycation inhibitors in combination are more effective in preventing such changes and could be an effective therapeutic option for preventing establishment of secondary complications of diabetes.
KeywordsProtein glycation Oxidative stress Fenton reaction Rotenone Aminoguanidine Vitamin E
Bovine serum albumin
- Vit E
Hepatocellular carcinoma cells
Advanced Glycation End-Products
Laxman Naghnath Bavkar acknowledges the University Grant Commission (UGC), New Delhi, India for a fellowship under the Special Assistance Program for Basic Scientific Research. Rahul Shivaji Patil acknowledges the Department of Science and Technology (DST), New Delhi, India for DST-INSPIRE fellowship. Chemicals and research work were supported by RUSA, Government of Maharashtra funded to Prof. (Mrs.) A. U. Arvindekar. The authors are thankful to the Department of Biochemistry, Shivaji University, Kolhapur for providing work place and laboratory.
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
- 5.Watkins NG, Thorpe SR, Baynes JW (1985) Glycation of amino groups in protein. Studies on the specificity of modification of RNase by glucose. J Biol Chem 260:10629–10636Google Scholar
- 12.Rival T, Page RM, Chandraratna DS, Sendall TJ, Ryder E, Liu B et al (2009) Fenton chemistry and oxidative stress mediate the toxicity of the beta-amyloid peptide in a Drosophila model of Alzheimer’s disease. Eur J Neurosci 29:1335–1347. https://doi.org/10.1111/j.1460-9568.2009.06701.x CrossRefGoogle Scholar
- 14.Li D, Devaraj S, Fuller C, Bucala R, Jialal I (1996) Effect of tocopherol on LDL oxidation and glycation: in vitro and in vivo studies. J Lipid Res 37:1978–1986Google Scholar
- 18.Nakamura A, Goto S (1996) Analysis of protein carbonyls with 2, 4-dinitrophenyl hydrazine and its antibodies by immunoblot in two-dimensional gel electrophoresis. J Biochem 119:768–774. https://doi.org/10.1093/oxfordjournals.jbchem.a021306 CrossRefGoogle Scholar
- 21.Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175Google Scholar
- 22.Beers RF, Sizer IW (1952) Spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195:133–140Google Scholar
- 33.Miyata T, Kurokawa K, De Strihou CVY (2000) Advanced glycation and lipoxidation end products: role of reactive carbonyl compounds generated during carbohydrate and lipid metabolism. J Am Soc Nephrol 11:1744–1752Google Scholar