BACKGROUND AND OBJECTIVE: The aim of the current study was to investigate in detail the effect of the active metabolite of vitamin D3 [1, 25 (OH)2 D3] in ameliorating the induced oxidative damage to DNA. MATERIALS AND METHODS: Primary cortical neuron cultures from one week old Wister rats were set up in sterile conditions. The neuron cultures were maintained for up to 72 h in culture in the presence of varying doses of vitamin D. Cells were exposed to (0.5 mM H2O2) for 2 h prior to collection of condition medium and cell pellet for Biochemical Assays. Control and H2O2 treated cultures were maintained without any treatment with vitamin D. RESULTS: Pre-treatment with 0.25 μg mL-1 for 24 and 48 h significantly reduced the oxidative stress. 8-hydroxydeoxyguanosine a ubiquitous marker of oxidative stress had also shown to be significantly reduced. The DNA damage marker PolyUB of histones was observed in the neuron treated with H2O2 only. CONCLUSION: This study revealed that oxidation of DNA by hydrogen peroxide caused extensive DNA damage, resulting in polyubiquitination of histones. The pre-treatment with vitamin D3 however completely reversed the DNA damage cascade induced by hydrogen peroxide and protected the DNA.
BACKGROUND AND OBJECTIVE: The aim of the current study was to investigate in detail the effect of the active metabolite of vitamin D3 [1, 25 (OH)2 D3] in ameliorating the induced oxidative damage to DNA. MATERIALS AND METHODS: Primary cortical neuron cultures from one week old Wister rats were set up in sterile conditions. The neuron cultures were maintained for up to 72 h in culture in the presence of varying doses of vitamin D. Cells were exposed to (0.5 mM H2O2) for 2 h prior to collection of condition medium and cell pellet for Biochemical Assays. Control and H2O2 treated cultures were maintained without any treatment with vitamin D. RESULTS: Pre-treatment with 0.25 μg mL-1 for 24 and 48 h significantly reduced the oxidative stress. 8-hydroxydeoxyguanosine a ubiquitous marker of oxidative stress had also shown to be significantly reduced. The DNA damage marker PolyUB of histones was observed in the neuron treated with H2O2 only. CONCLUSION: This study revealed that oxidation of DNA by hydrogen peroxide caused extensive DNA damage, resulting in polyubiquitination of histones. The pre-treatment with vitamin D3 however completely reversed the DNA damage cascade induced by hydrogen peroxide and protected the DNA.
Entities:
Keywords:
DNA damage; Oxidative stress; monoubiquitination; neurodegeneration; polyubiquitin (polyUb); vitamin D