南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (10): 1804-1809.doi: 10.12122/j.issn.1673-4254.2023.10.20

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转录组测序鉴定兔移植静脉桥血管再狭窄相关基因

周海深，杨利博，谢 诗，杨家明，张家庆   

  1. 南方医科大学珠江医院胸外科，广东 广州 510280
  • 出版日期:2023-10-20 发布日期:2023-11-03

Transcriptome sequencing for identification of the genes associated with restenosis of venous grafts in rabbits

ZHOU Haishen, YANG Libo, XIE Shi, YANG Jiaming, ZHANG Jiaqing   

  1. Department of Thoracic Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China
  • Online:2023-10-20 Published:2023-11-03

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摘要： 目的 使用转录组测序技术鉴定兔移植静脉桥血管再狭窄相关的基因。方法 选取40只实验用新西兰兔，分为实验组、对照组，上述两组再分为2周组和4周组，10只/组。实验组将兔左侧颈外静脉移植至左颈总动脉上，分别于2周、4周时取下桥血管标本，对不同组别标本进行病理学检查，观察血管内膜、中膜厚度，血管平滑肌细胞的增殖情况；对照组不做手术处理，2周或4周后取出左侧颈外静脉。之后使用转录组测序技术对术后4周的移植血管进行检测，与对照组对比，寻找差异表达的基因，并利用GO数据库及KEGG数据库对表达差异的基因进行筛选分析。结果 术后2周桥血管病理提示：内膜及中膜增厚，血管平滑肌细胞增殖，术后4周时，上述改变进一步加重。与对照组静脉相比，术后4周的移植静脉标本明显上调基因1583个，下调608个。用GO以及KEGG数据库分析筛选出前10个处于中心位置基因分别为CD4、ZAP70、SYK、CD28、PIK3CD、CXCR4、CCR5、ITK、CCL5、BTK。结论 CD4、ZAP70、SYK、CD28、PIK3CD、CXCR4、CCR5、ITK、CCL5和BTK可能是引起兔静脉桥血管再狭窄的基因。

关键词: 静脉移植；再狭窄；转录组；基因

Abstract: Objective To identify the genes associated with venous graft restenosis in rabbits using transcriptome sequencing. Methods Forty New Zealand rabbits were randomly divided into experimental group and control group, and in the experimental group, the left external jugular veins of the rabbits were engrafted to the left common carotid artery with continuous running suture; the rabbits in the control group received no operation. At 2 and 4 weeks after the operation, 10 rabbits from each group were euthanized and the venous grafts (in experimental group) or left external jugular vein (in control group) were harvested for measurement of the intima-media thickness using HE staining. RNA high- throughput sequencing (RNA-seq) was performed to identify the differentially expressed genes (DEGs) between the venous grafts and the control veins, and the biological functions of the DEGs were analyzed using GO and KEEG databases. Results In the experimental group, intima-media thickening with increased extracellular matrix and vascular smooth muscle cell proliferation occurred in the venous grafts at 2 weeks and aggravated at 4 weeks after the operation. RNA high-throughput sequencing identified 1583 up-regulated genes and 608 down-regulated genes in the venous grafts in the experimental group, and GO and KEGG analysis of the DEGs pinpointed 10 hub genes, namely CD4, ZAP70, SYK, CD28, PIK3CD, CXCR4, CCR5, ITK, CCL5 and BTK. Conclusion CD4, ZAP70, SYK, CD28, PIK3CD, CXCR4, CCR5, ITK, CCL5 and BTK are probably the key genes associated with vein graft restenosis in rabbits.

Key words: vein graft; restenosis; transcriptome; gene