微小RNA-1270靶向调控血管生成素样蛋白7抑制巨噬细胞炎症和脂质蓄积  

miR-1270-targeted regulation of angiopoietin-like protein 7 inhibits macrophage inflammation and lipid accumulation

作　　者：杜艾家 张曼[1] 陈鹤 王丽新 尚应殊 DU Aijia;ZHANG Man;CHEN He;WANG Lixin;SHANG Yingshu(Department of Cardiology,Central Hospital of Shenyang Medical College,Shenyang 110024,Liaoning,China)

机构地区：[1]沈阳医学院附属中心医院心血管内科,辽宁沈阳110024

出　　处：《山东大学学报(医学版)》2025年第2期1-9,20,共10页Journal of Shandong University:Health Sciences

基　　金：沈阳市科学技术计划项目(22-321-33-93)。

摘　　要：目的应用氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的小鼠巨噬细胞株制备动脉粥样硬化模型,观察微小RNA-1270(microRNA-1270,miRNA-1270)是否经血管生成素样蛋白7(angiopoietin-like protein 7,ANGPTL7)/p38途径干预巨噬细胞炎症及血脂代谢。方法培养小鼠单核巨噬细胞(RAW264.7),并加入ox-LDL构建动脉粥样硬化模型。根据干预条件不同分为空白组、ox-LDL组、ANGPTL7组、p38蛋白抑制组、p65蛋白抑制组、miR-1270模拟物组、miR-1270模拟物阴性对照组、miR-1270抑制物组和miR-1270抑制物阴性对照组。采用实时定量PCR法检测mRNA表达水平,采用Western blotting法检测蛋白质表达水平,采用油红染色法检测脂质蓄积。结果ox-LDL暴露的巨噬细胞中含红色脂肪微粒的巨噬细胞数量增加,ANGPTL7高表达,p38高表达,白介素-6(interleukin-6,IL-6)高表达,白介素-10(interleukin-10,IL-10)低表达,且ANGPTL7与p38及脂质蓄积呈正相关(P<0.01)。与ox-LDL组相比,ANGPTL7组ANGPTL7、p38及IL-6蛋白相对表达量增加,IL-10蛋白相对表达量减少,含红色脂肪微粒的巨噬细胞数量增加(P<0.01)。与ANGPTL7组相比,p38蛋白抑制组ANGPTL7和p38蛋白相对表达量无明显差异,IL-6蛋白相对表达量减少,IL-10蛋白相对表达量增加,含红色脂肪微粒的巨噬细胞数量减少(P<0.01)。与ANGPTL7组相比,p65蛋白抑制组各项指标表达量、含红色脂肪微粒的巨噬细胞数量差异无统计学意义(P>0.05)。与空白组相比,ox-LDL组miR-1270基因相对表达量减少,ANGPTL7基因及蛋白相对表达量增加,miR-1270与ANGPTL7基因相对表达量呈负相关(P<0.01)。与ox-LDL组相比,miR-1270模拟物组miR-1270基因相对表达量增加,ANGPTL7基因及蛋白相对表达量减少,p38及IL-6蛋白相对表达量减少,IL-10蛋白相对表达量增加,含红色脂肪微粒的巨噬细胞数量减少(P<0.01);miR-1270抑制物组miR-1270基因相对表达量减少,ANGPTL7基因及蛋白相对表�Objective To prepare an atherosclerosis(As)model employing an oxidised low-density lipoprotein(ox-LDL)-induced mouse macrophage cell line,so as to see whether microRNA-1270(miRNA-1270)interfered with macrophage inflammation and lipid metabolism via the angiopoietin-like protein 7(ANGPTL7)/p38 pathway.Methods Mouse mononuclear macrophages(RAW264.7)were cultured and ox-LDL was added to construct macrophage models.According to the different intervention conditions,the groups were as follows:blank group,ox-LDL group,ANGPTL7 group,p38 protein inhibition group,p65 protein inhibition group,miR-1270 mimic group,miR-1270 mimic negative control group,miR-1270 inhibitor group,and miR-1270 inhibitor negative control group.The mRNA expression level was detected by real-time quantitative PCR,the protein expression level was detected by Western bloting,and the lipid accumulation was detected by oil red staining.Results The number of macrophages containing red fat particles was increased in ox-LDL-exposed macrophages,with high expressions of ANGPTL7,p38,and IL-6,and low expression of IL-10,and a positive correlation between ANGPTL7 and p38 and lipid accumulation(P<0.01).Compared with the ANGPTL7 group,there was no significant difference in the relative expression of ANGPTL7 and p38 proteins in the p38 protein inhibition group,a decrease in the relative expression of IL-6 protein,an increase in the relative expression of IL-10 protein,and a decrease in the number of red adipose microparticle-containing macrophages(P<0.01).Compared with the ANGPTL7 group,there was no statistically significant difference in the expression of the indicators in the p65 protein inhibition group,and there was no statistically significant difference in the number of macrophages containing red fat particles(P>0.05).Compared with the blank group,the relative expression of miR-1270 gene was decreased and the relative expression of ANGPTL7 gene and protein was increased in the ox-LDL group,and miR-1270 was negatively correlated with the relative expressions

关 键 词：基因表观遗传学 动脉粥样硬化 微小RNA-1270 血管生成素样蛋白7 P38蛋白 炎症反应 脂质蓄积 

分 类 号：R543.5[医药卫生—心血管疾病]