中国血液净化

中国血液净化 ›› 2025, Vol. 24 ›› Issue (12): 1004-1009.doi: 10.3969/j.issn.1671-4091.2025.12.008

• 基础研究 • 上一篇    下一篇

线粒体氧化应激在小鼠动静脉内瘘内膜增生致狭窄中的作用

谢婷妃   钟晓玲   陈家辉   张艾莎   隋晓露   张燕子   许云鹏   陈继红   

  1. 518000 深圳，1深圳市宝安区人民医院/深圳大学第二附属医院/广东医科大学深圳宝安临床医学院肾内科
  • 收稿日期:2025-01-13 修回日期:2025-09-30 出版日期:2025-12-12 发布日期:2025-12-12
  • 通讯作者: 陈继红 E-mail:chenjihong0606@hotmail.com
  • 基金资助:
    深圳市宝安区高层次人才创新项目，深圳市宝安区科技计划基础研究项目(2022JD139)； 深圳市宝安区医学会医疗卫生科研项目(BAYXH2023008)； 深圳市宝安区引进高层次医学团队项目(202401)

Role of  mitochondrial oxidative stress in intimal hyperplasia-induced stenosis in a mouse arteriovenous fistula  model

XIE Ting-fei, ZHONG Xiao-ling, CHEN Jia-hui, ZHANG Ai-sha, SUI Xiao-lu, ZHANG Yan-zi, XU Yun-peng, CHEN Ji-hong   

  1. Department of Nephrology, The People’s Hospital of Baoan Shenzhen, The Second Affiliated Hospital of Shenzhen University, Shenzhen Baoan Clinical Medical School of Guangdong Medical University, Shenzhen 518000, China
  • Received:2025-01-13 Revised:2025-09-30 Online:2025-12-12 Published:2025-12-12
  • Contact: 518000 深圳，1深圳市宝安区人民医院/深圳大学第二附属医院/广东医科大学深圳宝安临床医学院肾内科 E-mail:chenjihong0606@hotmail.com

摘要： 目的  构建小鼠颈内动静脉内瘘(arteriovenous fistula,AVF)模型，筛选AVF内膜增生狭窄过程中的差异表达蛋白，分析研究参与AVF内膜增生狭窄的异常表达信号通路。 方法  采用C57BL/6小鼠，通过显微外科技术建立小鼠颈总动脉-颈外静脉AVF模型，分为假手术组(n=10)和AVF手术组(n=10)。评估AVF血管病理改变。应用串联质谱标签(tandem mass tag,TMT)定量蛋白组学分析2组差异表达的基因和蛋白，利用生物信息学方法对差异蛋白进行富集分析。采用人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)进行细胞实验分为对照组和剪切力组，检测活性氧和线粒体形态。 结果  动物实验，与假手术组相比，AVF手术组静脉新生内膜面积显著增生(t=6.469,P＜0.001)，由胶原纤维增生与肌层相互混杂构成。蛋白组分析共筛选出595个差异表达蛋白，富集分析表明显著差异蛋白主要参与线粒体稳态、能量代谢等生物过程，在糖脂代谢通路和过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptors,PPAR)通路中富集。细胞实验，剪切力组HUVECs中活性氧产生高于对照组(t=8.840,P＜0.001)，线粒体面积、线粒体长度低于对照组(t=5.926、4.579,均P＜0.001)，线粒体内超氧化物高于对照组(t=28.170,P＜0.001)。 结论  线粒体氧化应激损伤参与AVF内膜增生致狭窄过程。

关键词: 动静脉内瘘, 内膜增生, 蛋白组测序, 氧化应激

Abstract: Objective  To establish a mouse  model of internal carotid arteriovenous fistula (AVF) and screen for differentially expressed proteins during AVF intimal hyperplasia and stenosis, and analyze the aberrant signaling pathways associated with this process.  Methods  A common carotid artery-external jugular vein AVF model was established in C57BL/6 mice using microsurgical techniques. The mice were divided into a sham operation group (n=10) and an AVF operation group (n=10). Vascular pathological changes in the AVF were evaluated. Tandem mass tag (TMT) quantitative proteomics analysis was used to identify differentially expressed genes and proteins between the two groups, and bioinformatics methods were applied to perform enrichment analysis of the differentially expressed proteins. For the cell experiment, human umbilical vein endothelial cells (HUVECs) were divided into a control group and a shear stress group, to detect reactive oxygen species (ROS) and mitochondrial morphology.  Results  In the animal experiment, compared with the sham operation group, the AVF operation group exhibited significant venous intimal hyperplasia, characterized by a mixture of collagen fiber proliferation and muscular layer components. Proteomic analysis identified a total of 595 differentially expressed proteins. Enrichment analysis revealed that these genes were primarily involved in biological processes such as mitochondrial homeostasis and energy metabolism, and were enriched in glycolipid metabolism pathways and the peroxisome proliferators-activated receptors (PPAR) signaling pathway.  In the cell experiment, ROS production in HUVECs in the shear stress group was higher than in the control group (t=8.840, P＜0.001). Mitochondrial area and mitochondrial length were lower in the shear stress group compared to the control group (t=5.926, 4.579, both P＜0.001), while mitochondrial superoxide levels were higher  (t=28.170, P＜0.001).  Conclusion  Mitochondrial oxidative stress injury plays a role in the process of intimal hyperplasia and stenosis in AVF.

Key words: Arteriovenous fistula, Intimal hyperplasia, Proteome sequencing, Oxidative stress

中图分类号: