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Effects of Homocysteine on ERK Signaling and Cell Proliferation in Fetal Neural Stem Cells In Vitro

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Abstract

The aim of the present study was to determine if the excitatory amino acid homocysteine (Hcy) alters ERK signaling and cell proliferation in fetal neural stem cells (NSCs) in vitro. NSCs were isolated from fetal rats and grown in serum-free suspension medium. The cells were identified as NSCs by their expression of immunoreactive Sox2. NSCs were assigned to one of four treatment groups: vehicle control, low-dose Hcy group (Hcy-L, medium contained 30 μmol/L Hcy), middle-dose Hcy group (Hcy-M, 100 μmol/L Hcy) and high-dose Hcy group (Hcy-H, 300 μmol/L Hcy). Cell proliferation was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Protein expression levels of ERK1/2 and phosphorylated ERK1/2 were detected by Western blot. The effects of Hcy on NSC death, including apoptosis, were assessed by using flow cytometry and trypan blue exclusion. The results showed that NSCs grew as neurospheres in the serum-free medium. Hcy decreased ERK1/2 protein phosphorylation and NSC proliferation, but it did not induce cell death or apoptosis within the concentration from 30 to 300 μmol/L. The above results are consistent with the hypothesis that Hcy decreases fetal NSC proliferation by inhibiting ERK signaling.

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Acknowledgments

This project was supported by the National Natural Science Foundation of China (No. 81072289 and 30901192).

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Authors and Affiliations

  1. Department of Nutrition and Food Hygiene, School of Public Health, Tianjin Medical University, Tianjin, 300070, China

    Hai Yan, Xumei Zhang, Suhui Luo, Huan Liu, Xuan Wang & Guowei Huang

  2. Department of Cardiology, General Hospital of Tianjin Medical University, Tianjin, 300152, China

    Yuxia Gao

  3. Department of Exercise and Nutrition Sciences, School of Public Health and Health Professions, University at Buffalo, Buffalo, NY, 14214-8028, USA

    John X. Wilson

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  1. Hai Yan
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  2. Xumei Zhang
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  3. Suhui Luo
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  4. Huan Liu
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  5. Xuan Wang
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  8. Guowei Huang
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Correspondence to Guowei Huang.

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Yan, H., Zhang, X., Luo, S. et al. Effects of Homocysteine on ERK Signaling and Cell Proliferation in Fetal Neural Stem Cells In Vitro. Cell Biochem Biophys 66, 131–137 (2013). https://doi.org/10.1007/s12013-012-9461-z

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  • DOI: https://doi.org/10.1007/s12013-012-9461-z

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