半乳糖凝集素-3通过lncARSR影响小鼠冠状动脉粥样硬化易损斑块稳定的机制  

Balectin-3 affects the stability of vulnerable coronary atherosclerosis plaques by targeted modulation of lncARSR

作　　者：许名东 李丽华 李晚泉 刘家超 李小丹 叶浩彬 陈健芳 邹文毕 XU Mingdong;LI Lihua;LI Wanquan;LIU Jiachao;LI Xiaodan;YE Haobin;CHEN Jianfang;ZOU Wenhua(Sanshui Hospital Affiliated to Guangdong Medical University,Foshan 528100,China)

机构地区：[1]广东医科大学附属三水医院,广东佛山528100

出　　处：《南方医科大学学报》2021年第7期1067-1072,共6页Journal of Southern Medical University

基　　金：佛山市科技攻关项目(1920001000275)。

摘　　要：目的探讨半乳糖凝集素-3(Gal-3)通过lncARSR影响冠状动脉粥样硬化易损斑块稳定的机制。方法将60只雄性BALB/c小鼠随机分为3组(n=20):正常饮食组、高脂饮食组(15%脂肪+0.25%胆固醇饮食)、高脂饮食+lncARSR抑制剂组(15%脂肪和0.25%胆固醇饮食,隔日尾静脉注射50 nmol/L lncARSR抑制剂),12周后取材。通过苏丹IV和油红O染色分析小鼠冠状动脉粥样硬化及斑块损伤,Westen blot分析小鼠冠状动脉Gal-3和ARSR蛋白表达,免疫组化检测小鼠冠状动脉中PI3K和Akt表达。同时取冠状动脉组织进行细胞培养,分为对照转染组、Gal-3转染组和Gal-3抑制组,转染24 h后用双荧光素酶报告基因测定Gal-3与lncARSR的靶标关系,RT-qPCR检测转染细胞中肿瘤坏死因子-α(TNF-α)和白介素-β(IL-β)、IL-6的mRNA表达。结果正常饮食组苏丹IV阳性面积和红油染色面积以及ARSR的蛋白表达最低,而高脂饮食组最高(P<0.05);Gal-3转染组较对照转染组、Gal-3抑制组的WT-lncARSR酶活性升高(P=0.026),Gal-3抑制组较对照转染组WT-lncARSR酶活性降低(P=0.017)。高脂饮食组较正常饮食组PI3K和Akt的蛋白以及TNF-α、IL-β、IL-6的mRNA表达升高,高脂饮食+lncARSR抑制剂组较高脂饮食组PI3K和Akt的蛋白以及TNF-α、IL-β、IL-6的mRNA表达降低(P<0.05)。结论冠状动脉粥样硬化中Gal-3和lncARSR过量表达。Gal-3能靶向调控lncARSR,通过抑制lncARSR表达,调控PI3K/Akt信号通路,降低炎症反应,调节冠状动脉粥样硬化易损斑块稳定。Objective To investigate the mechanism by which galectin-3(Gal-3)affects the stability of vulnerable coronary atherosclerosis plaques through long non-coding RNA ARSR(lncARSR).Method Male BALB/c mice were randomly divided into normal diet group,high-fat diet group,high-fat diet+lncARSR inhibitor group(n=20).The high-fat diet contained 15%fat and 0.25%cholesterol,and lncARSR inhibitor was injected intravenously at 50 nmol/L every other day.After 12 weeks of high-fat diet feeding and treatment,the mice were euthanized for analyzing coronary atherosclerosis and plaque damage using Sudan IV and oil red O staining.The protein expressions of Gal-3 and ARSR in the coronary artery of the mice were analyzed with Western blotting,and the expressions of PI3 K and Akt were detected with immunohistochemistry.The coronary artery tissues were harvested from normal mice for cell culture,and the isolated cells were transfected with a Gal-3 mimic or a Gal-3 inhibitor.At 24 h after the transfection,dual luciferase reporter gene assay was performed to determine the target relationship between Gal-3 and lncARSR;the mRNA expressions of tumor necrosis factor(TNF-α),interleukin-β(IL-β)and IL-6 in the transfected cells were detected with RT-qPCR.Results The positively stained areas by Sudan IV and red oil O and the protein expression of lncARSR were the lowest in normal diet group and the highest in high-fat diet group(P<0.05).The protein expression of PI3 K and Akt and the mRNA expression of TNF-α,IL-βand IL-6 in high-fat diet group were higher than those in normal diet group.The protein expression of PI3 K and Akt and the mRNA expression of TNF-α,IL-βand IL-6 in high-fat diet+lncARSR inhibitor group were significantly lower than those in high-fat diet group(P<0.05).In the cell experiment,the activity of WT-lncARSR was significantly higher in Gal-3 mimic transfection group than in the control group and Gal-3 inhibition group(P=0.026),and was the lower in Gal-3 inhibition group than in the control group(P=0.017).Conclusion Gal-3 a

关 键 词：半乳糖凝集素-3 冠状动脉粥样硬化 lncARSR 斑块稳定 

分 类 号：R543.5[医药卫生—心血管疾病]