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首页> 外文期刊>Journal of applied toxicology >Effects of homocysteine on mesenchymal cell proliferation and differentiation during chondrogenesis on limb development
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Effects of homocysteine on mesenchymal cell proliferation and differentiation during chondrogenesis on limb development

机译：同型半胱氨酸对软骨发育过程中间充质细胞增殖和分化的影响

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摘要

High levels of homocysteine (Hcy) are related to an increased risk of the occurrence of congenital anomalies, including limb defects. However, few evaluations about how toxic levels of Hcy affect limb development have been reported. We investigated whether Hcy can affect the cell cycle proteins and proteins involved in mesenchymal cell differentiation during limb development, in a chicken embryo model. Embryos were treated with 20 mu mol d-l Hcy/50 mu l saline at embryonic day 2 and analyzed at embryonic day 6. Untreated control embryos received exclusively 50 mu l saline solution. To identify cells in proliferation and cell cycle proteins, as well as Pax1/9 and Sox9 proteins, we performed immunolocalization and flow cytometry analyses using the antibodies anti-phosphohistone H3, anti-p53, anti-p21, anti-proliferating cell nuclear antigen, anti-Pax1, anti-Pax9 and anti-Sox9. No significant differences in cell proliferation were observed between Hcy-treated and untreated embryos. We observed a decrease of the proliferating cell nuclear antigen and p21 proteins, both involved in the G(1) phase of cell cycle progression. On the other hand, in mesenchymal cells of the limbs, Hcy induces an increase of p53 protein, which can be activated by DNA damage. In cell differentiation, Hcy induced an increase mainly of Pax9 and Sox9 proteins. Our data indicate that the treatment with Hcy changes the mesenchymal cell dynamics during limb development, but does not change the morphology of the cartilage molds. These findings provide information to understand better the cellular basis of the toxicity of Hcy on chondrogenesis during limb development. Copyright (c) 2015 John Wiley & Sons, Ltd.
机译：高半胱氨酸（Hcy）水平与包括肢体缺损在内的先天性异常发生的风险增加有关。但是，关于Hcy的毒性水平如何影响肢体发育的评估报道很少。我们在鸡胚胎模型中研究了Hcy是否能影响细胞周期蛋白以及在肢体发育过程中参与间充质细胞分化的蛋白。在胚胎第2天用20μmold-1 Hcy /50μl盐水处理胚胎，并在胚胎第6天进行分析。未处理的对照胚胎仅接受50μl盐溶液。为了鉴定增殖和细胞周期蛋白以及Pax1 / 9和Sox9蛋白中的细胞，我们使用了抗磷酸化组蛋白H3，抗p53，抗p21，抗增殖细胞核抗原， anti-Pax1，anti-Pax9和anti-Sox9。在Hcy处理和未处理的胚胎之间，未观察到细胞增殖的显着差异。我们观察到增殖细胞核抗原和p21蛋白的减少，两者都参与细胞周期进程的G（1）阶段。另一方面，在四肢间质细胞中，Hcy诱导p53蛋白增加，而p53蛋白可以被DNA损伤激活。在细胞分化中，Hcy主要诱导Pax9和Sox9蛋白增加。我们的数据表明，Hcy处理可改变肢体发育过程中的间充质细胞动力学，但不会改变软骨模具的形态。这些发现为更好地了解Hcy对肢体发育过程中软骨形成毒性的细胞基础提供了信息。版权所有（c）2015 John Wiley＆Sons，Ltd.

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