iTRAQ蛋白质组学方法发现高同型半胱氨酸诱导的鸡胚神经管畸形中存在氧化磷酸化通路异常

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摘要目的筛选同型半胱氨酸硫代内酯(HTL)诱导鸡胚神经管畸形差异蛋白以明确高同型半胱氨酸(Hcy)导致神经管畸形的发病机制。方法采用白来航鸡种蛋于孵化28 h进行神经沟注射HTL处理,于第5天(E5)收集胚胎脑组织,采用iTRAQ 联合二维液相色谱-串联质谱技术 (2D-LC-MS/MS) 对差异蛋白质进行筛选,并进一步用蛋白质非标记 (1abel-free) 定量技术进行差异蛋白的验证。结果在鸡脑组织中共鉴定出369种差异蛋白质;KEGG通路分析提示差异蛋白富集于代谢通路和氧化磷酸化通路;1abel-free(PRM)验证氧化磷酸化通路蛋白ATP5H和ATP5MF在NTDs中表达异常。结论 HTL诱导鸡胚神经管畸形脑中氧化磷酸化通路蛋白异常表达。

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	张勤
	李丹
	白宝玲
	万春蕾
	肖宗慧

关键词 ： iTRAQ, 神经管畸形, 同型半胱氨酸硫代内酯, 鸡胚

Abstract：Objective To screen differentially expressed proteins of neural tube defects (NTDs) induced by homocysteine thiolactone (HTL) in chicken embryos and to elucidate the pathogenesis of NTDs induced by high Hcy . Methods White Leghorn chicken eggs were incubated for 28 hours and then were injected with HTL into nerve groove. Fetal brain tissues were collected in E5 day. Differential proteins were screened by iTRAQ combined with two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS), and further verified by 1abel-free quantitative technique. Results A total of 369 differentially expressed proteins were identified in brain tissues; KEGG pathway analysis indicated that differentially expressed proteins were enriched in metabolic pathways and oxidative phosphorylation pathways; PRM confirmed that the expression of oxidative phosphorylation pathway proteins ATP5H and ATP5MF were decrease in NTDs. Conclusion HTL induced abnormal expression of oxidative phosphorylation pathway proteins in chicken NTDs.

Key words： iTRAQ Neural tube defects HTL chicken embryos

收稿日期: 2019-08-29

基金资助:北京市自然科学基金(7182024)

引用本文:

张勤, 李丹, 白宝玲, 万春蕾, 肖宗慧. iTRAQ蛋白质组学方法发现高同型半胱氨酸诱导的鸡胚神经管畸形中存在氧化磷酸化通路异常[J]. 中国生育健康杂志, 2020, 31(3): 227-232.
ZHANG Qin, LI Dan, BAI Baoling, WAN Chunlei, XIAO Zonghui.. ITRAQ proteomics revealed that abnormal expressed protein of oxidative phosphorylation pathway was involved in the development of neural tube defects in chicken embryos. Chinese Journal of Reproductive Health, 2020, 31(3): 227-232.

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http://cjrh.bjmu.edu.cn/CN/ 或 http://cjrh.bjmu.edu.cn/CN/Y2020/V31/I3/227

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