靶向干预Sestrin 1对动脉粥样硬化小鼠斑块内巨噬细胞极化和血管内皮损伤的影响  

Impact of targetedly intervening Sestrin 1 on intra-plaque macrophage polarization and vascular endothelial injury in atherosclerotic mice

作　　者：肖彭卓 杨克平 XIAO Peng-zhuo;YANG Ke-ping(Medical College,Yangtze University,Jinzhou 434023,China;Jinzhou Central Hospital/the Second Medical College of Yangtze University,Jinzhou 434000,China)

机构地区：[1]长江大学医学院,湖北省荆州市434023 [2]湖北省荆州市中心医院/长江大学第二临床医学院,荆州市434000

出　　处：《广西医学》2021年第11期1340-1345,共6页Guangxi Medical Journal

摘　　要：目的分析Sestrin 1对动脉粥样硬化小鼠斑块内巨噬细胞极化的调控作用及对血管内皮炎性损伤的影响。方法将40只ApoE^(-/-)小鼠随机分为对照组、模型组、si-NC组和si-Sestrin 1组,对照组小鼠正常饲养,其余小鼠均采用高脂饮食法建立动脉粥样硬化模型。建模成功2周后,si-NC组和si-Sestrin 1组分别给予尾静脉注射携带对照序列和Sestrin 1干扰序列的RAW 264.7细胞,其他两组输入等量细胞培养液。末次输注后次日,分离各组小鼠胸主动脉,油红O染色后测量斑块总面积,并测定斑块内活性氧活性;分离斑块中的巨噬细胞,采用流式细胞仪测定巨噬细胞表面标志物CD64、CD197、CD163、CD206阳性率,实时定量PCR测定肿瘤坏死因子α、白细胞介素(IL)-6、IL-1β、诱导型一氧化氮合酶(iNOS)、IL-10、转化生长因子β(TGF-β)的mRNA表达量,蛋白免疫印迹法测定Sestrin 1蛋白表达量。结果(1)对照组、模型组、si-Sestrin 1组的斑块面积依次增加,斑块内活性氧活性依次升高,Sestrin 1蛋白表达量依次降低(均P<0.05),但si-NC组上述指标与模型组比较差异无统计学意义(均P>0.05)。(2)对照组、模型组、si-Sestrin 1组M1型巨噬细胞的表面标志物CD64和CD197阳性率、iNOS mRNA表达量依次升高,M2型巨噬细胞表面标志物CD163和CD206阳性率、IL-10和TGF-βmRNA表达量依次降低(均P<0.05),但si-NC组上述指标与模型组比较差异无统计学意义(均P>0.05)。结论下调Sestrin 1可促进动脉粥样硬化斑块内巨噬细胞向M1型的极化,并由此参与血管内皮细胞的炎性损伤,这可能是它参与动脉粥样硬化进展的重要机制之一。Objective To analyze the regulatory effect of Sestrin 1 on the polarization of intra-plaque macrophages and its impact on vascular endothelial inflammatory injury in atherosclerotic mice.Methods Forty ApoE^(-/-)mice were randomly divided into control group,model group,si-NC group,and si-Sestrin 1 group.Mice in the control group received normal feeding,while mice in the remaining groups received high-fat diet to establish an atherosclerosis model.Two weeks after successful modeling,the si-NC and si-Sestrin 1 groups were injected with RAW 264.7 cells carrying control sequences and Sestrin 1 interfere sequences,respectively,through caudal vein,and the other two groups were infused with isometric cell culture solution.On the next day after final infusion,the thoracic aorta of mice in all groups were separated,then the total area of plaque was measured after oil red O staining,and the reactive oxygen species(ROS)activity in the plaques was determined.The macrophages were removed from plaque,then the positive rates of macrophage surface markers CD64,CD197,CD163 and CD206 were determined by flow cytometry,the mRNA expression of tumor necrosis factorα,interleukin(IL)-6,IL-1β,inducible nitric oxide synthase(iNOS),IL-10 and transforming growth factorβ(TGF-β)was determined by real-time quantitative PCR,and the protein expression of Sestrin 1 was determined by Western blotting.Results(1)The plaque area and intra-plaque ROS activity increased,and Sestrin 1 protein expression decreased in the control group,model group and si-Sestrin 1 group successively(all P<0.05),but there was no statistically significant difference in the aforementioned indices between the si-NC group and the model group(all P>0.05).(2)The positive rates of M1-type macrophage surface markers(CD64 and CD197)and the iNOS mRNA expression increased,while the positive rates of M2-type macrophage surface markers(CD163 and CD206)and the expression of IL-10 and TGF-βmRNAs declined in the control group,model group and si-Sestrin 1 group successively(all P<0.05)

关 键 词：动脉粥样硬化 Sestrin 1 巨噬细胞极化 血管内皮 炎性损伤 小鼠 

分 类 号：R543.5[医药卫生—心血管疾病]