氟中毒及补钙干预大鼠骨骼差异蛋白的分离与鉴定
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国家自然科学基金 (No. 31001089),山西省自然科学基金 (No. 2014011027-3) 资助。


Separation and identification of differential protein in rat’s bone with fluorosis and calcium supplementation intervention
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National Natural Science Foundation of China (No. 31001089), National Science Foundation of Shanxi Province (No. 2014011027-3).

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    摘要:

    为从蛋白水平探索钙缓解氟中毒的作用机制,本试验建立氟中毒和补钙干预动物模型进行骨骼差异蛋白的分离鉴定。通过双向凝胶电泳技术(Two-dimensional electrophoresis,2-DE) 和基质辅助质量飞行时间质谱技术 (MALDI-TOF MS) 鉴定骨骼中的差异蛋白质,并通过gene ontology (GO) 注释、pathway富集和互作网络对差异蛋白进行分析。结果表明,与对照组 (C) 相比,在氟中毒组 (F) 和氟中毒补钙组 (F+Ca) 中有17种差异性的蛋白,包括Ⅰ型胶原蛋白 (Col1a1)、肌动蛋白 (Actb)、蛋白质谷氨酰胺转移酶2 (Tgm2) 等。这些差异蛋白富集到黏着斑 (Focal adhesion)、PI3K-Akt信号通路、AMPK信号通路等38条骨代谢通路。并且这些蛋白的功能主要与细胞骨架、能量代谢、物质转运、离子通道、细胞凋亡有关。因此推测,钙可能通过调控黏着斑、PI3K-Akt、AMPK等信号通路进而缓解氟对骨骼带来的损伤,具体作用机制还需要进一步验证。

    Abstract:

    In order to explore the mechanisms underlying the calcium alleviating fluorosis at protein level, we made an attempt to establish fluorosis and calcium supplementation rat models to isolate and identify bone differential proteins. The bone proteins of different groups were compared by two-dimensional electrophoresis (2-DE) and mass spectrometry (MALDI-TOF MS), and analyzed by gene ontology annotation, pathway enrichment and interaction networks. The 17 proteins were identified in the fluorosis group (F) and the fluorosis calcium supplement group (F+Ca), including type I collagen (Col1a1), actin (Actb), protein glutamine transferase 2 (Tgm2), compared with the control group (C). These differential proteins are enriched in 38 bone metabolic pathways such as focal adhesion, PI3K-Akt signaling pathway, and AMPK signaling pathway. And the functions of these proteins are mainly related to cytoskeleton, energy metabolism, substance transport, ion channel, and apoptosis. Therefore, it is speculated that calcium may alleviate the fluoride-induced bone damage by regulating the focal adhesion, PI3K-Akt, AMPK and other signaling pathway, but the specific mechanism needs further research.

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王金明,徐慧淼,闫子鹏,杨镓蓉,朱雅雅,成小芳,王俊东. 氟中毒及补钙干预大鼠骨骼差异蛋白的分离与鉴定[J]. 生物工程学报, 2019, 35(6): 1097-1108

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  • 收稿日期:2018-11-29
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  • 在线发布日期: 2019-06-21
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