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Mangiferin Reduced Etoposide-induced DNA Damage in Mononuclear Human Umbilical Cord Blood Cells Via Activating Nrf2-ARE Signaling

Author: LiZuoZuo
Tutor: ChenYan
School: Huazhong University of Science and Technology
Course: Department of Hematology
Keywords: Mangiferin Nrf2 ARE NQO1 DNA damage Human umbilical cord blood cells
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Objective: In this experiment, normal human cord blood mononuclear cells as experimental subjects , umbilical cord blood cells DNA damage induced by chemical mutagens VP-16 in the in vitro studies mangiferin (MA) whether preventive protective role , and Nrf2/ARE antioxidant pathway in anti- carcinogens link between the role of the DNA damage , and further explore prevent leukemia Mangiferin possible molecular mechanisms . Methods : In vitro separation of normal cord blood mononuclear cell DNA damage model induced by VP-16 , the mangiferin early intervention in the umbilical cord blood cells that divided into VP-16 and MA VP- 16 groups. , Respectively, using the single cell gel electrophoresis experiment that comet experiment SCGE ( comet assay ) and micronucleus test (MNT) to detect DNA damage ; indirect immunofluorescence ( confocal microscopy) and immunoblotting (Western Blotting) detection Nrf2 positioning mangiferin role of Nrf2 and NQO1 protein expression ; gel electrophoretic mobility shift assay (EMSA) detection of nuclear Nrf2 ARE combination of cis-acting elements . Results: chemical mutagen VP-16 in human umbilical cord blood cells have a significant role in DNA damage 10μg/μl , 2h can render the whole-cell DNA damage , the extent of the damage was time and concentration-dependent ; the Mangiferin intervention group and not the plus Mangiferin group comparison , mangiferin group ( is early intervention 4h ) DNA damage extent than not low plus Mangiferin group ; either in the umbilical cord blood cells in the control group or the intervention group in MA , Nrf2 are mainly located in the nucleus . Mangiferin - induced a Nrf2 expression of rendering time and concentration -dependent nuclear Nrf2 ARE specific binding nucleus cis-acting element induced ARE downstream detoxification gene NQO1 expression , also has a time-dependent manner , which exerts its anti- activity of detoxification oxide . Conclusion : In this experiment , confirmed Mangiferin inhibit and protection of the umbilical cord blood cells DNA damage induced by chemical mutagens VP-16 , this protective mechanism may activating Nrf2/ARE antioxidant pathway to complete . Therefore Mangiferin early intervention activity to protect cells from DNA damage may be making it a new tumor drug prevention , in particular, is to provide a preventive therapy related leukemia may be worthy of further study .

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