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Effect of glucose-mediated LDL oxidation on the P388D1 macrophage-like cell line
S A Millican1, M Bagga, R Eddy
1University of Cambridge, Department of Pathology, UK.
Abstract:
Oxidised human low density lipoprotein (LDL) is thought to play a role in the development of atherosclerosis. Recent reports suggest that glucose-derived oxidants are capable of oxidising LDL. In this report, the effect of glucose-mediated oxidation of LDL upon the macrophage like cell line, P388D(1), was examined. Glucose-mediated oxidation of LDL was assessed by changes in the electrophoretic mobility of LDL and by analysis of lipid content using gas chromatography. The presence of Cu(II) (0.5 microM) was essential for the oxidation of LDL. The oxidation was potentiated by glucose in a dose- and time-dependent manner. At the concentration of LDL used (1 mg/ml), high concentrations of glucose (up to 500 mM) were required to oxidise LDL. The electrophoretic mobility of LDL correlated with the degree of lipid oxidation; both correlated with an inhibitory effect of oxidised LDL upon P388D(1) DNA synthesis. Diethylenetriaminepentaacetic acid (DETAPAC), a transition metal chelator, and aminoguanidine (AMG), an anti-glycation agent, inhibited the oxidation of LDL and attenuated the effects on DNA synthesis. Thus, glucose can mediate transition metal-dependent oxidation of LDL to a level that can affect P388D(1) cells, a mechanism which might have relevance to accelerated atherosclerosis in diabetic patients.
Insights
High glucose levels can oxidize low-density lipoprotein (LDL), potentially contributing to atherosclerosis. This glucose-mediated LDL oxidation impacts cell DNA synthesis, suggesting a link to diabetic complications.
Area of Science:
- Biochemistry
- Cell Biology
- Pathophysiology
Background:
- Oxidized low-density lipoprotein (LDL) is implicated in atherosclerosis development.
- Glucose-derived oxidants may contribute to LDL oxidation.
Purpose of the Study:
- To investigate the effect of glucose-mediated LDL oxidation on the P388D(1) macrophage cell line.
- To explore the mechanism by which glucose influences LDL oxidation and its cellular impact.
Main Methods:
- Assessed LDL oxidation via electrophoretic mobility and lipid content analysis (gas chromatography).
- Examined the influence of glucose concentration, time, and copper(II) on LDL oxidation.
- Evaluated the effect of oxidized LDL on P388D(1) cell DNA synthesis.
- Investigated the role of transition metal chelators (DETAPAC) and anti-glycation agents (aminoguanidine).
Main Results:
- Copper(II) was essential for LDL oxidation, potentiated by glucose in a dose- and time-dependent manner.
- High glucose concentrations (up to 500 mM) were required to oxidize LDL (at 1 mg/ml).
- LDL oxidation correlated with inhibited P388D(1) DNA synthesis.
- DETAPAC and aminoguanidine inhibited LDL oxidation and attenuated the effects on DNA synthesis.
Conclusions:
- Glucose can mediate transition metal-dependent oxidation of LDL.
- This glucose-mediated LDL oxidation can affect macrophage-like cells (P388D(1)).
- The findings suggest a potential mechanism for accelerated atherosclerosis in diabetic patients.