Effect of Si-Miao-Yong-An decoction on the differentiation of monocytes, macrophages, and regulatory T cells in ApoE-/- mice
Effect of Si-Miao-Yong-An decoction on the differentiation of monocytes, macrophages, and regulatory T cells in ApoE-/- mice
复制标题
四妙永安汤对ApoE(-/-)小鼠单核细胞、巨噬细胞和调节性T细胞分化的影响
DOI：
10.1016/j.jep.2021.114178
复制
发表时间：
2021-05-12
影响因子：
5.4
通讯作者：
Zhang, Jun-Ping
中科院分区：
文献类型：
--
作者：
Chen, Xin-Nong;Ge, Qi-Hui;Zhang, Jun-Ping
民族药理学相关性：四妙永安汤（SMYAD）是一个著名的传统中药配方。《医史秘录》是清代医学家陈士铎主编的《医史秘录》中的记载。SMYAD传统上用于治疗血栓闭塞性脉管炎。目前主要用于心血管疾病的临床应用和研究。研究目的：探讨参麦益气汤对载脂蛋白E基因敲除（ApoE- / -）小鼠动脉粥样硬化（AS）病变及单核细胞、巨噬细胞和调节性T细胞（Treg细胞）分化的影响。材料与方法：8只C57 BL/6 J小鼠，正常饮食喂养16周作为对照组。将40只ApoE-/-小鼠随机分为模型组、阿托伐他汀组、四味地黄汤低剂量组、四味地黄汤中剂量组和四味地黄汤高剂量组。ApoE- /-小鼠用西方饮食（WD）喂养8周，并连续给药8周。采用酯酶法测定血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）水平。HE染色观察主动脉窦形态学变化，油红O染色观察主动脉及主动脉窦脂质浸润情况，计算脾脏指数。流式细胞术检测脾脏中Ly 6C高和Ly 6C低单核细胞亚群、巨噬细胞及其M1表型以及Treg细胞的比例。免疫组织化学染色检测主动脉窦内分化抗原36（CD 36）、清道夫受体A1（SRA 1）、凝集素样氧化低密度脂蛋白受体1（LOX-1）、F4/80和叉头框架蛋白3（FOXP 3）的表达。采用酶联免疫吸附法（ELISA）检测血清氧化低密度脂蛋白（ox-LDL）、白细胞介素-113（IL-113）、IL-18、转化生长因子-13（TGF-13）和IL-10水平。结果如下：与模型组比较，四味养阴汤组血清TC、LDL-C水平降低，主动脉窦病理改变减轻，主动脉及主动脉窦脂质浸润减少。这些降低伴随着CD 36、SRA 1和LOX-1的显著下调。此外，脾脏中Ly 6Chigh促炎单核细胞亚群、巨噬细胞及其M1表型的比例显著降低，而Treg细胞的比例增加。此外，在主动脉窦中，F4/80的表达减少的同时，FOXP 3的表达增加。血清促炎因子IL-113和IL-18水平降低。结论：参麦益气汤能改善AS相关的病理变化，并能抑制WD饮食诱导的Apoe- /-小鼠的脂质沉积。可能的机制是抑制单核细胞和巨噬细胞的分化和募集，促进Treg细胞的分化和募集，以及减少促炎因子的分泌。
Ethnopharmacological relevance: Si-Miao-Yong-An decoction (SMYAD) is a renowned traditional Chinese medicinal formula. SMYAD was originally recorded in the "Shi Shi Mi Lu", which was edited by medical scientist Chen Shi'duo during the Qing Dynasty. SMYAD has been traditionally used to treat thromboangiitis obliterans. At present, it is mainly used in clinical applications and research of cardiovascular diseases. Aim of the study: To explore the effects of SMYAD on the pathological changes of atherosclerosis (AS) and the differentiation of monocytes, macrophages, and regulatory T (Treg) cells in apolipoprotein E knockout (ApoE- / -) mice. Materials and methods: Eight C57BL/6J mice, which were fed with normal diet for 16 weeks, were used as control group. Forty ApoE-/- mice were randomly divided into model group, atorvastatin group, SMYAD low-dose (SMYAD-LD) group, SMYAD medium-dose (SMYAD-MD) group, and SMYAD high-dose (SMYAD-HD) group. ApoE- /- mice were fed with western diet (WD) for 8 weeks, and the drugs were continuously administered for 8 weeks. The levels of serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were measured by the esterase method. Morphological changes of the aortic sinus in mice were observed by hematoxylin-eosin (HE) staining, the lipid infiltration of the aorta and aortic sinus were observed by oil red O staining, and the spleen index was calculated. The proportion of Ly6Chigh and Ly6Clow monocyte subsets, macrophages, and their M1 phenotype, as well as Treg cells in spleen were measured by flow cytometry. The expressions of cluster of differentiation 36 (CD36), scavenger receptor A1 (SRA1), lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), F4/80, and fork head frame protein 3 (FOXP3) in aortic sinus were assessed by immunohistochemical staining. The serum levels of oxidized low density lipoprotein (ox-LDL), interleukin-113 (IL-113), IL-18, transforming growth factor-13 (TGF-13), and IL-10 were measured by enzyme-linked immunosorbent assays (ELISA). Results: Compared with the model group, the level of serum TC and LDL-C decreased in the SMYAD group, the pathological changes of aortic sinus decreased, and lipid infiltration of aorta and aortic sinus also decreased. These decreases were accompanied by a significant downregulation of CD36, SRA1, and LOX-1. Furthermore, the proportions of Ly6Chigh pro-inflammatory monocyte subsets, macrophages, and their M1 phenotypes in spleen decreased significantly, while the proportion of Treg cells increased. In addition, while the expression of F4/80 decreased, the expression of FOXP3 increased in the aorta sinus. The levels of serum pro-inflammatory factors IL113 and IL-18 decreased. Conclusions: SMYAD can improve the pathological changes associated with AS and can inhibit lipid deposition in ApoE- /- mice induced by WD diet. The likely mechanism is the inhibition of the differentiation and recruitment of monocytes and macrophages, the promotion of the differentiation and recruitment of Treg cells, as well as the reduction of the secretion of pro-inflammatory factors.