Expression changes of DNA methyltransferase in microglia after acute hypoxia
Objective:To investigate the changes of DNA methyltransferase in microglia after hypoxia.Methods:BV2 cells were used as the research object,and they were divided into the control group,H-0 group(hypoxia for 12 h without reoxygenation),H-2 group(hypoxia for 12 h with reoxygenation for 2 h),H-4 group(hypoxia for 12 h with reoxygenation for 4 h),H-8 group(hypoxia for 12 h with reoxygenation for 8 h),H-12 group(hypoxia for 12 h with reoxygenation for 12 h),and H-24 group(hypoxia for 12 h with reoxygenation for 24 h).The BV2 cells were cultured under hypoxia condition for 12 h and reoxygenated at different time points to construct the model,and the cell morphology was observed in the bright field by fluorescence inverted microscope.The expression changes of DNMT1,DNMT 3A,and DNMT 3B at the mRNA level were detected by Real-time PCR,and the expression changes of DNMT1,DNMT 3A,and DNMT 3B at the protein level were detected by Western blot.Results:Compared to the Control group,the H-0 group had a decreased number of cells,wrinkled cell morphology,and elongated tentacles in most cells.The cells were dull and the refractive index was reduced.With the reoxygenation time increasing(H-2,H-4,H-8,H-12,H-24),the adherent cells gradually de-creased.Compared with the control group,the protein expression level of DNMT1 was significantly increased in the H-0 group(P＜0.05),and the mRNA and protein expression level of DNMT1 was significantly decreased in the H-8,H-12,and H-24 groups(P＜0.05).mRNA and protein expression level of DNMT 3 A was significantly decreased in the H-8,H-12 groups(P＜0.05).mRNA and protein expression of DNMT 3B in cells of H-0,H-4,H-8,H-12,and H-24 groups did not change significantly.Conclusion:DNMT1 and DNMT3A play important roles in microglia after hypoxia.
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