亚精胺/精胺N1-乙酰转移酶1参与血管平滑肌细胞钙化  

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

作　　者：王凯茹 付士哲 李家辉 燕茹[1,3,4,5] 马玉茹[1,3,4,5] 师波 冶从燕 闫瑞 丛广志[1,3,4,5] 贾绍斌[1,3,4,5] Wang Kairu;Fu Shizhe;Li Jiahui;Yan Ru;Ma Yuru;Shi Bo;Ye Congyan;Yan Rui;Cong Guangzhi;Jia Shaobin(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;Department of Cardiology,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学总医院医学科学研究院,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 [3]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,宁夏回族自治区银川市750004 [4]宁夏医科大学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 [5]宁夏医科大学总医院心内科,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2025年第32期6836-6842,共7页Chinese Journal of Tissue Engineering Research

基　　金：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~~。

摘　　要：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/Lβ-甘油磷酸钠和3 mmol/L氯化钙的DMEM培养基诱导细胞钙化,50,100μmol/L二乙酰胺三氮脒组在钙化培养基的基础上,用50,100μmol/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,50,100μmol/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�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 singlecell 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/Lβ-glycerophosphate sodium and 3 mmol/L calcium chloride,and the 50,100μmol/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 Runtrelated 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 phosphata

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 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R543.5