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Role of N-(carboxymethyl)lysine in the development of ischemic heart disease in type 2 diabetes mellitus.

A Ahmed K, Muniandy S, S Ismail I - J Clin Biochem Nutr (2007)

Bottom Line: Serum CML levels were measured by enzyme-linked immunosorbent assay using polyclonal anti-CML antibodies.In conclusion, this study demonstrates the effect of both diabetes and oxidative stress on the higher levels of circulating CML.These results showed that increased serum levels of CML are associated with the development of IHD in Type 2 diabetes mellitus.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
This study aims to determine the levels of N(epsilon)-(carboxymethyl)lysine (CML) in patients with Type 2 diabetic patients with and without ischemic heart disease (IHD) and to find for a possible association between circulating CML and a number of clinical parameters including lipids, hemoglobin A1c (HbA1c) and malondialdehyde (MDA) in Type 2 diabetic IHD patients. Serum CML levels were measured by enzyme-linked immunosorbent assay using polyclonal anti-CML antibodies. Serum levels of CML and MDA were assessed in 60 IHD patients with Type 2 diabetes, 43 IHD patients without Type 2 diabetes, 64 Type 2 diabetics without IHD, and 80 sex- and age-matched healthy subjects. Correlations studies between CML levels and lipids, HbA1c, and lipid peroxidation were performed in Type 2 diabetes patients with and without IHD. A statistical significance was observed in the levels of serum glucose, lipids (triglyceride, total cholesterol, HDL-cholesterol), MDA, HbA1c, CML and LDL-cholesterol (p<0.05) between the groups of the study. CML levels were significantly increased in diabetic IHD patients compared with Type 2 diabetes patients but without IHD (537.1 +/- 86.1 vs 449.7 +/- 54.9, p<0.001). A positive correlation was observed between serum levels of CML and MDA, r = 0.338 (p = 0.008) in Type 2 diabetes patients with IHD. However, age, HbA1c and lipids had no significant influence on CML levels among diabetics (p>0.05). In conclusion, this study demonstrates the effect of both diabetes and oxidative stress on the higher levels of circulating CML. These results showed that increased serum levels of CML are associated with the development of IHD in Type 2 diabetes mellitus.

No MeSH data available.


Related in: MedlinePlus

Agarose gel electrophoresis of glycated and nonglycated proteins. Native BSA (lane 1), CML-BSA (lane 2), CEL-BSA (lane 3), CEL-Hb (lane 4), CML-Hb (lane 5) and Hb (lane 6). Samples in loading buffer (bromophenol blue, 1.5 M Tris and 20% glycerol, pH 9.2) were loaded into agarose gel (1.5%) in 0.1 M Tris and 0.038 M glycine buffer (pH 8.7). The gel tray was run for 40–60 min at 100 V. The gel was stained with 1% Coomassie blue (1 h) followed by overnight destaining.
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Figure 1: Agarose gel electrophoresis of glycated and nonglycated proteins. Native BSA (lane 1), CML-BSA (lane 2), CEL-BSA (lane 3), CEL-Hb (lane 4), CML-Hb (lane 5) and Hb (lane 6). Samples in loading buffer (bromophenol blue, 1.5 M Tris and 20% glycerol, pH 9.2) were loaded into agarose gel (1.5%) in 0.1 M Tris and 0.038 M glycine buffer (pH 8.7). The gel tray was run for 40–60 min at 100 V. The gel was stained with 1% Coomassie blue (1 h) followed by overnight destaining.

Mentions: Agarose gel electrophoresis was used to evaluate the purity of the modified proteins employing 1.5% agarose in Tris-glycine buffer. Figure 1 showed that CML- and CEL-glycated proteins preparations revealed a faster anodic migration than native BSA or Hb, indicating that significant glycation had occurred. Quantitatively, the modified proteins were subjected to TNBS assay to estimate the amount of glycation. The degree of glycation was calculated from the reduction of absorbance compared with native proteins and expressed as percentage of modification. The percentages of lysine residues modifications in CML-BSA, CEL-BSA, CML-Hb, and CEL-Hb CML- and CEL-glycated proteins versus total lysine residues in the native proteins are 66, 48, 62, and 39%, respectively.


Role of N-(carboxymethyl)lysine in the development of ischemic heart disease in type 2 diabetes mellitus.

A Ahmed K, Muniandy S, S Ismail I - J Clin Biochem Nutr (2007)

Agarose gel electrophoresis of glycated and nonglycated proteins. Native BSA (lane 1), CML-BSA (lane 2), CEL-BSA (lane 3), CEL-Hb (lane 4), CML-Hb (lane 5) and Hb (lane 6). Samples in loading buffer (bromophenol blue, 1.5 M Tris and 20% glycerol, pH 9.2) were loaded into agarose gel (1.5%) in 0.1 M Tris and 0.038 M glycine buffer (pH 8.7). The gel tray was run for 40–60 min at 100 V. The gel was stained with 1% Coomassie blue (1 h) followed by overnight destaining.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2170948&req=5

Figure 1: Agarose gel electrophoresis of glycated and nonglycated proteins. Native BSA (lane 1), CML-BSA (lane 2), CEL-BSA (lane 3), CEL-Hb (lane 4), CML-Hb (lane 5) and Hb (lane 6). Samples in loading buffer (bromophenol blue, 1.5 M Tris and 20% glycerol, pH 9.2) were loaded into agarose gel (1.5%) in 0.1 M Tris and 0.038 M glycine buffer (pH 8.7). The gel tray was run for 40–60 min at 100 V. The gel was stained with 1% Coomassie blue (1 h) followed by overnight destaining.
Mentions: Agarose gel electrophoresis was used to evaluate the purity of the modified proteins employing 1.5% agarose in Tris-glycine buffer. Figure 1 showed that CML- and CEL-glycated proteins preparations revealed a faster anodic migration than native BSA or Hb, indicating that significant glycation had occurred. Quantitatively, the modified proteins were subjected to TNBS assay to estimate the amount of glycation. The degree of glycation was calculated from the reduction of absorbance compared with native proteins and expressed as percentage of modification. The percentages of lysine residues modifications in CML-BSA, CEL-BSA, CML-Hb, and CEL-Hb CML- and CEL-glycated proteins versus total lysine residues in the native proteins are 66, 48, 62, and 39%, respectively.

Bottom Line: Serum CML levels were measured by enzyme-linked immunosorbent assay using polyclonal anti-CML antibodies.In conclusion, this study demonstrates the effect of both diabetes and oxidative stress on the higher levels of circulating CML.These results showed that increased serum levels of CML are associated with the development of IHD in Type 2 diabetes mellitus.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
This study aims to determine the levels of N(epsilon)-(carboxymethyl)lysine (CML) in patients with Type 2 diabetic patients with and without ischemic heart disease (IHD) and to find for a possible association between circulating CML and a number of clinical parameters including lipids, hemoglobin A1c (HbA1c) and malondialdehyde (MDA) in Type 2 diabetic IHD patients. Serum CML levels were measured by enzyme-linked immunosorbent assay using polyclonal anti-CML antibodies. Serum levels of CML and MDA were assessed in 60 IHD patients with Type 2 diabetes, 43 IHD patients without Type 2 diabetes, 64 Type 2 diabetics without IHD, and 80 sex- and age-matched healthy subjects. Correlations studies between CML levels and lipids, HbA1c, and lipid peroxidation were performed in Type 2 diabetes patients with and without IHD. A statistical significance was observed in the levels of serum glucose, lipids (triglyceride, total cholesterol, HDL-cholesterol), MDA, HbA1c, CML and LDL-cholesterol (p<0.05) between the groups of the study. CML levels were significantly increased in diabetic IHD patients compared with Type 2 diabetes patients but without IHD (537.1 +/- 86.1 vs 449.7 +/- 54.9, p<0.001). A positive correlation was observed between serum levels of CML and MDA, r = 0.338 (p = 0.008) in Type 2 diabetes patients with IHD. However, age, HbA1c and lipids had no significant influence on CML levels among diabetics (p>0.05). In conclusion, this study demonstrates the effect of both diabetes and oxidative stress on the higher levels of circulating CML. These results showed that increased serum levels of CML are associated with the development of IHD in Type 2 diabetes mellitus.

No MeSH data available.


Related in: MedlinePlus