南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (7): 1067-1072.doi: 10.12122/j.issn.1673-4254.2021.07.15

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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

许名东，李丽华，李晚泉，刘家超，李小丹，叶浩彬，陈健芳，邹文毕   

  • 出版日期:2021-07-20 发布日期:2021-07-19

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

  • Online:2021-07-20 Published:2021-07-19

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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。

关键词: 半乳糖凝集素-3；冠状动脉粥样硬化；lncARSR；斑块稳定

Abstract: 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 PI3K 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 PI3K and Akt and the mRNA expression of TNF-α, IL-β and IL-6 in high-fat diet group were higher than those in normal diet group. The protein expression of PI3K and Akt and the mRNA expression of TNF-α, IL-β and 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 and lncARSR are overexpressed in coronary atherosclerosis. Through a mechanism for targeted inhibition of lncARSR, Gal-3 regulates the PI3K/Akt signaling pathway to suppress inflammation and thus regulate the stability of vulnerable coronary atherosclerosis plaques.

Key words: galectin-3; coronary atherosclerosis; lncARSR; plaque stability