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Folic acid inhibits homocysteine-induced cell apoptosis in human umbilical vein endothelial cells

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Abstract

Homocysteine may be responsible for vascular endothelial cell injury, which occurs early in the pathology of cardiovascular disease. Homocysteine metabolism requires enzymatic interaction with vitamins such as folic acid, vitamin B12, and vitamin B6. We hypothesized that folic acid alleviated homocysteine-induced vascular injury by regulating the metabolic pathway of apoptosis. Human umbilical vein endothelial cells were incubated for 48 h with folic acid at the concentrations of 0–1000 nmol/L, in combination with either 1000 μmol/L homocysteine or vehicle for the first 24 h. We then assessed cell viability and apoptosis by methyl thiazolyl tetrazolium assay and flow cytometry, respectively. To further investigate how folic acid influenced cell apoptosis, we also analyzed the activities of caspase-3/7 and the mRNA and protein expressions of BCL2, BAX, TP53, CASP3, and CASP8 in human umbilical vein endothelial cells. We showed that folic acid increased cell viability and decreased apoptosis in a dose-dependent manner, and that this effect was mediated by decreased caspase-3/7 activity, upregulated BCL2/BAX ratio, and downregulated TP53, CASP3, and CASP8 expressions. Thus, we conclude that folic acid inhibits cell apoptosis and ameliorates homocysteine toxicity by regulating the expression of apoptosis-related genes in human umbilical vein endothelial cells.

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Abbreviations

AMV:

Avian myeloblastosis virus

ANOVA:

Analysis of variance

BCL2:

B-cell lymphoma 2

BAX:

Bcl-2-associated protein X

Caspase:

Cysteinyl aspartate-specific proteinase

Hcy:

Homocysteine

HUVEC:

Human umbilical vein endothelial cells

HRP:

Horseradish peroxidase

MTT:

Methyl thiazolyl tetrazolium

PCR:

Polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel

TP53:

Tumor suppressor p53

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Acknowledgements

This research was supported by a grant from the National Natural Science Foundation of China (No. 81373002).

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Authors and Affiliations

  1. Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin, 300070, China

    Shanshan Cui, Wen Li, Pengyan Wang, Xin Lv & Guowei Huang

  2. Department of Cardiology, General Hospital of Tianjin Medical University, Tianjin, 300052, China

    Yuxia Gao

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  1. Shanshan Cui
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  2. Wen Li
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Correspondence to Yuxia Gao or Guowei Huang.

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Cui, S., Li, W., Wang, P. et al. Folic acid inhibits homocysteine-induced cell apoptosis in human umbilical vein endothelial cells. Mol Cell Biochem 444, 77–86 (2018). https://doi.org/10.1007/s11010-017-3232-5

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