Comparative Study Of Native And Fructose Glycated Human Placental DNA

Abstract

Objective : The aim of our study was to glycate human placental DNA with Fructose and conduct a comparative study of properties of native and glycated DNA on the basis of UV spectrometry, fluorescence, and agarose gel electrophoresis. Methodology : Human placental DNA (10g/ml) was incubated with 25mM fructose for 5, 10 and 15 days in phosphate buffer .Absorption profile and fluorescence emission spectra of native and glycated DNA samples were recorded. Native and glycated DNA was run on 0.8% agarose gel and electrophoresis was conducted for 2 hours at 30 mA. The gels were stained with Ethidium bromide (0.5 g/ml) and viewed under UV illumination and photographed. Results : DNA incubated with 25mM fructose for 5 days generated appreciable amount of DNA- AGE (as yellow products in the assay tube). On spectrophotometric analysis, Native DNA gave a peak at 260nm but in glycated DNA, a peak at 260nm and a new peak around 360nm were seen. On Agarose gel electrophoresis, native DNA showed a pattern typical of genomic DNA and glycated DNA showed decreased ethidium assisted fluorescence. Conclusion : Fructose glycates Human Placental DNA resulting in formation of strand breaks and modification of bases.