Effects of folic acid and vitamin B12 on human umbilical vein endothelial cells against homocysteine-induced apoptosis |
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| Authors: | MA Peng-jun ZHANG Ming-hao GUO Wei MA Sheng-chao SUN Lei LI Gui-zhong JIANG Yi-deng |
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| Institution: | 1. College of Clinical Medicine, Ningxia Medical University, Yinchuan 750004, China;
2. Ningxia Laboratory of Vascular Injury and Repair Research, Ningxia Medical University, Yinchuan 750004, China;
3. Department of Preclinical Medicine College, Ningxia Medical University, Yinchuan 750004, China |
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| Abstract: | AIM: To investigate the effect of folic acid and vitamin B12 on homocysteine (Hcy)-induced apoptosis of human umbilical vein endothelial cells (HUVECs) through mammalian sterile 20-like kinase 1 (MST1). ME-THODS: HUVECs were cultured in the absence (control group), or presence of 100 μmol/L Hcy alone (Hcy group) or 100 μmol/L Hcy plus 30 μmol/L folic acid and vitamin B12 (intervention group) for 72 h. The effect of Hcy on the apoptosis of HUVECs was analyzed by flow cytometry. The transfection efficiency of DNA methyltransferase 1 (DNMT1)-overexpressing adenovirus was observed under fluorescence inverted microscope. The mRNA and the protein levels of DNMT1 and MST1 were determined by RT-qPCR and Western blot. The DNA methylation level of MST1 promoter was detected by methylation-specific PCR. RESULTS: Compared with control group, the apoptotic rate (P<0.01) and the expression of MST1 at mRNA (P<0.01) and protein (P<0.05) levels in the HUVECs were significantly increased, while the mRNA levels of DNMT1 was decreased in Hcy group (P<0.01). In addition, folic acid and vitamin B12 treatment significantly inhibited Hcy-mediated apoptosis of HUVECs (P<0.01), increase in MST1 mRNA level (P<0.01) and decrease in DNMT1 mRNA level (P<0.01). Meantime, the mRNA level of MST1 was positively correlated with the apoptotic rate of the HUVECs (r=0.943 9, P<0.001). The expression of DNMT1 at mRNA and protein levels was significantly increased after the transfection of DNMT1-overexpressing adenovirus into HUVECs (P<0.01), and a large amount of green fluorescent protein expression was observed. Meanwhile, the DNA methylation level of MST1 promoter was increased (P<0.01), while the protein level of MST1 was decreased (P<0.01).CONCLUSION: Up-regulation of MST1 promotes Hcy-induced apoptosis of HUVECs, while folic acid and vitamin B12 exert an anti-apoptosis effect, which might be regulated by hypermethylation of MST1 promoter region. |
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| Keywords: | Homocysteine Folic acid Vitamin B12 Mammalian sterile 20-like kinase 1 DNA methylation |
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