黄芪甲苷通过调控miR-33a及ABCA1信号发挥抗动脉粥样硬化作用  

Anti-atherosclerosis effect of astragaloside is related to miR-33a/ABCA1 signaling

作　　者：秦合伟 李彦杰[1] 张志鑫 任锟 李斯锦 卢永保 QIN He-wei;LI Yan-jie;ZHANG Zhi-xin;REN Kun;LI Si-jin;LU Yong-bao(Henan Province Hospital of TCM, Second Affiliated Hospital of Henan University of TCM, Zhengzhou 450002, China;Henan University of TCM, Zhengzhou 450002, China)

机构地区：[1]河南省中医院,河南中医药大学第二附属医院,河南郑州450002 [2]河南中医药大学,河南郑州450002

出　　处：《中国病理生理杂志》2018年第12期2120-2126,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81704030);河南省高等学校重点科研项目计划(No.18A360008);河南省中医药科学研究专项课题(No.2017ZY2067);河南省科技攻关计划项目(No.182102311158);河南省中医院专项课题(No.2017YJKT02);河南省中医临床学科领军人才培养计划(No.2100202).

摘　　要：目的:观察黄芪甲苷是否通过调控微小RNA-33a(miR-33a)而影响下游信号三磷酸腺苷结合盒转运体A1(ABCA1)的表达,促进巨噬细胞内胆固醇流出,从而研究黄芪甲苷抗动脉粥样硬化的作用机制。方法:体内实验:采用HE染色检测各组小鼠主动脉横截面病理损伤程度real-time PCR和Western blot分别检测小鼠主动脉ABCA1的mRNA和蛋白表达。体外实验:建立THP-1巨噬细胞源性泡沫细胞,然后用黄芪甲苷含药SD大鼠血清处理,采用real-time PCR检测细胞miR-33a表达;细胞随机分为空白血清组、黄芪甲苷含药血清组及黄芪甲苷含药血清+miR-33a mimic处理组,real-time PCR和Western blot法检测细胞ABCA1的mRNA和蛋白表达,油红O染色和高效液相色谱法检测细胞内脂质含量,[3H]法检测细胞内胆固醇流出。结果:体内实验显示,黄芪甲苷组小鼠血管各层结构正常,排列整齐,局部有小灶性的钙化颗粒物沉积,病变轻,斑块小,泡沫细胞和脂质减少,弹力板基本完整,病变程度明显轻于模型组。与模型组相比,黄芪甲苷组小鼠主动脉的miR-33a表达量降低,ABCA1 mRNA和蛋白相对表达量均升高(P <0. 05)。体外实验显示,在不影响THP-1巨噬细胞源性泡沫细胞活性的情况下,黄芪甲苷含药血清能明显上调ABCA1的mRNA和蛋白表达,但能被转染miR-33 mimic抑制;黄芪甲苷含药血清可减少细胞中的脂质蓄积,但能被细胞中转染miR-33 mimic所弱化;黄芪甲苷减少细胞内胆固醇蓄积与其促进细胞内胆固醇流出有关,细胞中转入过量miR-33a可以抑制胆固醇流出。结论:黄芪甲苷可通过减少miR-33a的生成,进而上调ABCA1表达,促进巨噬细胞中胆固醇流出,这可能是黄芪甲苷抗动脉粥样硬化作用的分子机制之一。AIM:To study whether astragaloside affects the expression of ATP binding cassette transporter A1(ABCA1)by regulating miR-33a and promotes the outflow of cholesterol in macrophages.METHODS:In the in vivo experiments,HE staining was used to detect the pathological damage of the cross section of aorta in the mice.The expression of ABCA1at mRNA and protein levels in mouse aorta was determined by real-time PCR and Western blot.In the in vitro experiments,THP-1macrophage-derived foam cells were established and then treated with astragaloside-containing serum.Real-time PCR was used to detect the expression of miR-33a.The cells were randomly divided into blank serum group,astragaloside serum group and astragaloside serum+miR-33a mimic group.The expression of ABCA1at mRNA and protein levels was determined by real-time PCR and Western blot.Oil red O staining and high-performance liquid chromatography were used to detect intracellular lipid content.The method of[3H]incorporation was used to detect intracellular cholesterol outflow.RESULTS:In vivo experiments showed that the blood vessels of the mice in astragaloside group were structurally normal,with neat arrangement,localized small calcified particles,mild lesions,small plaques,reduced foam cells and li-pid,and basically complete elastic plates,indicating that the pathological changes were significantly lighter than those in model group.Compared with model group,the expression of miR-33a in the aorta of the mice in astragaloside group was decreased and the relative expression of ABCA1at mRNA and protein levels was increased(P<0.05).In vitro experiments showed that astragaloside significantly up-regulated the expression of ABCA1at mRNA and protein levels,but this effect was inhibited by the transfection of miR-33mimic without affecting the cell viability.Astragaloside reduced the lipid accumulation in the cells,but this effect was attenuated by miR-33mimic.Astragaloside reduced intracellular cholesterol accumulation in relation to its promotion of intracellular cholesterol

关 键 词：黄芪甲苷 微小RNA-33a 三磷酸腺苷结合盒转运体A1 动脉粥样硬化 脂质代谢 

分 类 号：R543.5[医药卫生—心血管疾病] R285.5[医药卫生—内科学]