中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (32): 6836-6842.doi: 10.12307/2025.941

• 血管组织构建 vascular tissue construction • 上一篇    下一篇

亚精胺/精胺N1-乙酰转移酶1参与血管平滑肌细胞钙化

王凯茹1，2，3，4，付士哲1，2，3，4，李家辉1，3，4，燕  茹1，3，4，5，马玉茹1，3，4，5，师  波1，2，3，4，冶从燕1，2，3，4，闫  瑞1，2，3，4， 丛广志1，3，4，5，贾绍斌1，3，4，5   

  1. 宁夏医科大学总医院，1医学科学研究院，5心内科，宁夏回族自治区银川市  750004；2宁夏医科大学第一临床医学院，宁夏回族自治区银川市  750004；宁夏医科大学，3国家卫生健康委员会代谢性心血管疾病研究重点实验室，4血管损伤与修复研究重点实验室，宁夏回族自治区银川市  750004
  • 收稿日期:2024-10-14 接受日期:2024-12-10 出版日期:2025-11-18 发布日期:2025-04-25
  • 通讯作者: 贾绍斌，博士生导师，教授，主任医师，宁夏医科大学总医院，心内科，医学科学研究院，宁夏回族自治区银川市 750004；宁夏医科大学，国家卫生健康委员会代谢性心血管疾病研究重点实验室、血管损伤与修复研究重点实验室，宁夏回族自治区银川市 750004 共同通讯作者：丛广志，博士，硕士生导师，副主任医师，宁夏医科大学总医院，心内科，医学科学研究院，宁夏回族自治区银川市 750004；宁夏医科大学，国家卫生健康委员会代谢性心血管疾病研究重点实验室、血管损伤与修复研究重点实验室，宁夏回族自治区银川市 750004
  • 作者简介:王凯茹，女，1994年生，山东省菏泽市人，汉族，宁夏医科大学在读硕士，主要从事血管钙化方面的研究。
  • 基金资助:
    国家自然科学基金项目(82260086)，项目负责人：贾绍斌；宁夏心脏瓣膜疾病规范化诊疗技术应用示范(2022FRD05046)，项目负责人：贾绍斌

Spermidine/spermine N1-acetyltransferase 1 participates in vascular smooth muscle cell calcification

Wang Kairu1, 2, 3, 4, Fu Shizhe1, 2, 3, 4, Li Jiahui1, 3, 4, Yan Ru1, 3, 4, 5, Ma Yuru1, 3, 4, 5, Shi Bo1, 2, 3, 4, Ye Congyan1, 2, 3, 4, Yan Rui1, 2, 3, 4, Cong Guangzhi1, 3, 4, 5, Jia Shaobin1, 3, 4, 5   

  1. 1Institute of Medical Research, 5Department of Cardiology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2First Clinical Medical College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Key Laboratory of Metabolic Cardiovascular Disease Research, National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 4Key Laboratory of Vascular Injury and Repair Research, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2024-10-14 Accepted:2024-12-10 Online:2025-11-18 Published:2025-04-25
  • Contact: Jia Shaobin, Doctoral supervisor, Professor, Chief physician, Department of Cardiology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Institute of Medical Research, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Metabolic Cardiovascular Disease Research, National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Vascular Injury and Repair Research, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Co-corresponding author: Cong Guangzhi, PhD, Master’s supervisor, Associate chief physician, Department of Cardiology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Institute of Medical Research, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Metabolic Cardiovascular Disease Research, National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Vascular Injury and Repair Research, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Wang Kairu, Master candidate, Institute of Medical Research, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; First Clinical Medical College, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Metabolic Cardiovascular Disease Research, National Health Commission, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Vascular Injury and Repair Research, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82260086 (to JSB); Ningxia Heart Valve Disease Standardized Diagnosis and Treatment Technology Application Demonstration, No. 2022FRD05046 (to JSB) 

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摘要：

文题释义：

血管钙化：是指血管壁异常沉积钙盐的病理过程，与多种心血管疾病密切相关。正常情况下，血管平滑肌细胞维持血管结构和功能，然而在病理条件下，血管平滑肌细胞会发生表型转换，从收缩型转变为类骨细胞型，这种病理性钙化过程导致血管弹性降低、僵硬度增加。
生物信息学：是结合生物学、计算机科学和统计学的交叉学科。它主要用于分析和解释大量生物数据，如基因组序列、蛋白质结构等，应用范围包括基因组学、蛋白质组学和系统生物学。

背景：多胺在组织钙化中发挥重要作用。亚精胺/精胺N1-乙酰转移酶1作为调控细胞内多胺代谢的关键限速酶，其异常表达与多种病理过程相关，但在血管钙化中的作用尚未明确。
目的：探讨亚精胺/精胺N1-乙酰转移酶1在大鼠血管平滑肌细胞钙化中的作用。
方法：①生物信息学分析：通过GEO数据集分析亚精胺/精胺N1-乙酰转移酶1在人颈动脉粥样硬化斑块及其周围健康颈动脉组织中的表达差异；通过PanglaoDB公共基因表达数据库单细胞测序观察亚精胺/精胺N1-乙酰转移酶1细胞群的表达丰度和定位。②将大鼠血管平滑肌细胞分为3组，对照组用DMEM培养基培养，钙化组用含10 mmol/L β-甘油磷酸钠和3 mmol/L氯化钙的DMEM培养基诱导细胞钙化，50，
100 μmol/L二乙酰胺三氮脒组在钙化培养基的基础上，用50，100 μmol/L亚精胺/精胺N1-乙酰转移酶1抑制剂二乙酰胺三氮脒处理细胞。培养7-10 d，进行茜素红S染色以及细胞钙含量和碱性磷酸酶活性检测，Western blot检测Runt相关转录因子2、骨形态发生蛋白2、α-平滑肌肌动蛋白和亚精胺/精胺N1-乙酰转移酶1的蛋白表达，免疫荧光染色检测Runt相关转录因子2和亚精胺/精胺N1-乙酰转移酶1的表达。
结果与结论：①生物信息学分析显示，与健康颈动脉组织相比，颈动脉粥样硬化斑块中亚精胺/精胺N1-乙酰转移酶1和Runt相关转录因子2的表达显著上调 (P < 0.05)；单细胞测序数据库分析证实了亚精胺/精胺N1-乙酰转移酶1在血管平滑肌细胞中表达；②与对照组比较，钙化组Runt相关转录因子2、骨形态发生蛋白2、亚精胺/精胺N1-乙酰转移酶1、钙含量和碱性磷酸酶活性明显增加，而α-平滑肌肌动蛋白表达明显减少(均P < 0.05)；与钙化组比较，50，100 μmol/L二乙酰胺三氮脒组Runt相关转录因子2、骨形态发生蛋白2、钙含量和碱性磷酸酶活性明显降低，而α-平滑肌肌动蛋白表达明显增加 (均P < 0.05)；③免疫荧光实验显示，与钙化组比较，50，100 μmol/L二乙酰胺三氮脒组Runt相关转录因子2的表达强度明显降低。结果证实，亚精胺/精胺N1-乙酰转移酶1可能通过上调Runt相关转录因子2的表达，促进血管平滑肌细胞钙化。  

https://orcid.org/0009-0008-3106-0598(王凯茹)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 大鼠血管平滑肌细胞, 亚精胺/精胺N1-乙酰转移酶1(SAT1), Runt相关转录因子2, 血管钙化, 生物信息学, 平滑肌肌动蛋白

Abstract: BACKGROUND: Polyamines play a crucial role in tissue calcification. Spermidine/spermine N1-acetyltransferase 1(SAT1), as a key rate-limiting enzyme regulating intracellular polyamine metabolism, has been associated with various pathological processes. However, its role in vascular calcification remains unclear.
OBJECTIVE: To investigate the role of SAT1 in rat vascular smooth muscle cell calcification. 
METHODS: (1) Bioinformatics analysis: Differential expression of SAT1 in human carotid atherosclerotic plaques and their surrounding healthy carotid artery tissues were using GEO datasets. PanglaoDB database was used to analyze SAT1 expression abundance and localization across different cell types through single-cell sequencing. (2) Rat vascular smooth muscle cells were divided into three groups: a control group cultured in DMEM medium, a calcification group induced by DMEM medium containing 10 mmol/L β-glycerophosphate sodium and 3 mmol/L calcium chloride, and the 50, 100 μmol/L diacetylaminotriazamidine groups treated with the SAT1 inhibitor, diacetylaminotriazamidine, in addition to the calcification medium. After 7-10 days of culture, alizarin red S staining was performed, and cellular calcium content and alkaline phosphatase activity were assessed. Western blot was used to detect the protein expression of Runt-related transcription factor 2, bone morphogenetic protein 2, alpha-smooth muscle actin, and SAT1. Immunofluorescence staining was conducted to examine the expression of Runt-related transcription factor 2 and SAT1.   
RESULTS AND CONCLUSION: (1) Bioinformatics analysis revealed significantly upregulated expression of SAT1 and Runt-related transcription factor 2 (P < 0.05) in carotid atherosclerotic plaques compared with healthy carotid tissues (P < 0.05). Single-cell sequencing database analysis confirmed SAT1 expression in vascular smooth muscle cells. (2) Compared with the control group, the calcification group showed significantly increased Runt-related transcription factor 2, bone morphogenetic protein 2, SAT1, calcium content, and alkaline phosphatase activity, while alpha-smooth muscle actin expression was significantly decreased (all P < 0.05). Compared with the calcification group, the 50 and 100 μmol/L diacetylaminotriazamidine groups showed significantly decreased Runt-related transcription factor 2, bone morphogenetic protein 2, calcium content, and alkaline phosphatase activity, while alpha-smooth muscle actin expression was significantly increased (all P < 0.05). (3) Immunofluorescence experiments demonstrated that compared with the calcification group, the expression intensity of Runt-related transcription factor 2 was significantly reduced in the 50 and 100 μmol/L diacetylaminotriazamidine groups. Overall, SAT1 may promote vascular smooth muscle cell calcification by upregulating Runt-related transcription factor 2 expression.  

Key words: rat vascular smooth muscle cells, spermidine/spermine N1-acetyltransferase 1 (SAT1), Runt-related transcription factor 2, vascular calcification, bioinformatics, smooth muscle actin

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