摘要
目的:通过观察血管软化丸是否通过调控miR-33a,进而影响下游信号ABCA1的表达,促进巨噬细胞内胆固醇流出,研究血管软化丸抗动脉粥样硬化的作用机制。方法:通过病理学检测观察血管软化丸对动脉粥样硬化的影响,通过体外实验建立THP-1巨噬细胞源性泡沫细胞,然后用血管软化丸含药血清处理,Real-Time PCR技术检测细胞miR-33a表达。细胞随机分为空白对照组、血管软化丸含药血清处理组、血管软化丸含药血清+miR-33a mimic处理组,RT-PCR和Western blotting法检测细胞ABCA1 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,各中药组血管各层结构正常,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,病变程度明显低于空白对照组。与空白对照组相比,血管软化丸高、中、低三个剂量组小鼠主动脉的miR-33a表达量降低、ABCA1基因和蛋白相对表达量均升高,差异有统计学意义(P<0.05)。体外实验显示,在不影响细胞活性状况下,血管软化丸含药血清呈浓度和时间依赖性下调miRNA33a表达;血管软化丸含药血清能明显上调ABCA1 mRNA和蛋白的表达,但能被转染miRNA33mimic抑制;血管软化丸含药血清可减少THP-1巨噬细胞源性泡沫细胞中的脂质蓄积,但能被细胞中转染miRNA33 mimic所弱化;EGCG减少细胞内胆固醇蓄积是与其促进细胞内胆固醇流出有关,细胞中转入过量miRNA33a可以抑制胆固醇流出。结论:血管软化丸可通过减少miRNA33a的生成,进而上调ABCA1表达,促进巨噬细胞中胆固醇流出,这可能是血管软化丸抗动脉粥样硬化作用的分子机制之一。
Objective： To study the anti - atherosclerosis mechanism of Vascular Softening Pill by observing whether it can regulate miR -33a to influence the expression of ABCA1, and promote cholesterol efflux from macrophagocyte. Methods： Pathological examination was used to observe the effects of Vascular Softening Pill on atherosclerosis. THP - 1 macrophage - derived foam cells were established by in vitro experiment. The cells were intervened with serum containing Vascular Softening Pill. lit - PCtl was applied to detect the expression of mill-33a. The cells were randomly divided into three groups, i.e. group A（blank control） , group B （serum containing Vascular Softening Pill） , and group C （serum containing Vascular Softening Pill ＋ mill-33a mimic treatment）. Protein and mRNA expressions of ABCA1 were tested by RT- PCll and Western blotting method; intracellular lipid content was detected by Red O staining and HPLC; intracellular cholesterol efflux was examined by [ 3 H 1 method. Results： In vivo experiment, blood vessels were normally and neatly arranged in all TCM groups, with focal deposition of calcified particles; the lesions were mild, the patches were small, the foam cells and the lipids were reduced, the elastic plates were basically complete, and the pathological changes were less than those of the control group. The expression of mill -33a in the aor- ta was significantly decreased ; however, protein and mRNA expressions of ABCA1 were significantly increased in the Vascular Softening Pill groups of high - dose, medium - dose, and low - dose, compared to those of the control group （P 〈 0.05 ）. In vitro experiments, serum containing Vascular Softening Pill down - regulated the expression of miRNA33a in a dose - and - time - dependent manner; it significantly up - regulated protein and mllNA expressions of ABCA1, which could be inhibited by transfected miRNA33 mimic; it reduced lipid accumulation of THP -1 macrophage -derived foam cells, which could be weakened by transfected miRNA33 m
作者
秦合伟
李彦杰
任锟
张志鑫
邢若星
卢永保
QIN Hewei;LI Yanjie;REN Kun;ZHANG Zhixin;XING Ruoxing;LU Yongbao(Henan Province Hospital of TCM,the Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,China)
出处
《中医药信息》
2018年第6期1-7,共7页
Information on Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(No.81704030)
河南省高校重点科研项目(No.18A360008)
河南省中医药科学研究专项课题(No.2017ZY2067)
河南省中医院专项课题(No.2017YJKT02)
