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山东大学学报 (医学版) ›› 2026, Vol. 64 ›› Issue (8): 48-55.doi: 10.6040/j.issn.1671-7554.0.2025.0847

• 基础医学 • 上一篇    

AC16心肌细胞缺氧/复氧损伤后N-糖基化蛋白质组学分析

任妍欣1,2,黄庆文2,贾文娟2,杨军1,2   

  1. 1.滨州医学院第二临床医学院, 山东 烟台 264003;2.烟台毓璜顶医院心内科, 山东 烟台 264001
  • 发布日期:2026-08-14
  • 通讯作者: 杨军. E-mail:yangjyhd@163.com
  • 基金资助:
    山东省自然科学基金(ZR2023QH348)

N-glycoproteomic analysis of AC16 cardiomyocytes after hypoxia/reoxygenation injury

REN Yanxin1,2, HUANG Qingwen2, JIA Wenjuan2, YANG Jun1,2   

  1. 1. Second Clinical Medical College of Binzhou Medical University, Yantai 264003, Shandong, China;
    2. Department of Cardiology, Yantai Yuhuangding Hospital, Yantai 264001, Shandong, China
  • Published:2026-08-14

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摘要： 目的 基于质谱技术的定量N-糖蛋白质组学,对正常培养的AC16心肌细胞和缺氧/复氧(hypoxia/reoxygenation, H/R)处理后的AC16心肌细胞进行分析,从而为心肌缺血再灌注损伤(myocardial ischemia reperfusion injury, MIRI)的临床诊疗提供新靶点。 方法 采用缺氧培养箱对细胞进行H/R处理后,采用定量N-糖蛋白质组学质谱技术鉴定对照组与H/R组差异表达糖蛋白,通过生物信息学分析筛选关键调控分子,并通过蛋白质印迹实验验证核心靶点的表达特征。 结果 蛋白质组学鉴定到698种蛋白,其中66种显著上调(含42种差异糖基化蛋白,186个修饰位点),29种显著下调(含7种差异糖基化蛋白,27个修饰位点)。通路富集分析显示差异蛋白主要参与PI3K/AKT信号通路(P<0.05)和ECM-受体互作(P<0.05)。机制研究发现分化群44(cluster of differentiation 44, CD44)以及低氧上调蛋白 1(hypoxia up-regulated 1, HYOU1)在H/R模型中呈现显著表达上调(P<0.05)。 结论 本研究分析出AC16细胞H/R损伤中发生改变的 N-糖蛋白及 N-糖基化位点的系统层面视图。发现与MIRI相关的一系列具有异常 N-糖基化水平的糖蛋白,包括 CD44、HYOU1,将为糖基化表位的精准干预或为心肌保护提供新型治疗策略。

关键词: AC16心肌细胞, 心肌缺血再灌注损伤, 蛋白质组学, N糖基化, 质谱分析

Abstract: Objective To profile the quantitative N-glycoproteome of AC16 cardiomyocytes under normoxia and following hypoxia/reoxygenation(H/R)via mass spectrometry-based glycoproteomics, aiming to identify novel therapeutic targets for myocardial ischemia-reperfusion injury(MIRI). Methods After subjecting the cells to hypoxia/reoxygenation treatment, quantitative N-glycoproteomics mass spectrometry was employed to identify differentially expressed glycoproteins between the control group and the H/R group. Bioinformatics analysis was performed to screen for key regulatory molecules. The expression profiles of core candidate targets were subsequently validated by Western blotting analysis. Results Proteomics identified 698 proteins, including 66 upregulated(42 differentially glycosylated, 186 modification sites)and 29 downregulated(7 differentially glycosylated, 27 sites). Pathway analysis showed these proteins were mainly involved in the PI3K/AKT signaling pathway(P<0.05)and ECM-receptor interaction(P<0.05). Mechanistic studies found cluster of differentiation 44(CD44)and hypoxia up-regulated 1(HYOU1)to be significantly upregulated in the H/R model(P<0.05). Conclusion The study offers a systems-level view of altered N-glycoproteins and glycosylation sites in AC16 cells after H/R injury. It discovered a series of glycoproteins with abnormal N-glycosylation levels related to MIRI, such as CD44 and HYOU1. These findings may offer novel treatment strategies for targeted glycotope intervention and myocardial protection.

Key words: AC16 cardiomyocyte, Myocardial ischemia-reperfusion injury, Proteomics, N-glycosylation, Mass spectrometry analysis

中图分类号: 

  • R33
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