Shenzhen Center for Disease Control and Prevention, Shenzhen, 518055, Guangdong, China.
School Hygiene Division, Shenzhen Center for Disease Control and Prevention, Shenzhen, 518055, Guangdong, China.
Biomed Pharmacother. 2019 Oct;118:109332. doi: 10.1016/j.biopha.2019.109332. Epub 2019 Aug 22.
Atherosclerosis (AS), a severe disease characterized by an accumulation of lipids and fibers in the large arteries, is the most important contributor to ischemic stroke (IS). Although microRNAs (miRNAs) have been found in circulating blood, the role of miRNAs in the progression of AS remains unknown. In a previous study, we demonstrated that the spleen tyrosine kinase (Syk) gene plays a vital role in the process of IS. In the present study, we aimed to clarify whether the miRNAs targeting the Syk gene might slow the development of AS. Candidate miRNAs were screened in U937 and THP-1 cells via Bioinformatics analyses, RT-qPCR and dual-luciferase reporter assay. ApoE mice were used as an AS animal model. RAAV transfection was performed to identify the roles of Syk gene and miRNAs in the development of AS in ApoE mice. HE staining, Oil red O staining and immunohistochemistry were used to determine the mechanism of AS. RT-qPCR and western blotting were performed to determine the expressions of miRNAs and proteins, respectively. Over-expression of the Syk gene accelerated the development of AS. miR-377 effectively mediated the expression of the Syk gene in vitro and in vivo experiments. Further analysis indicated that over-expression of miR-377 partly alleviated the development of AS by down-regulating the expression of the Syk gene. This study identifies a novel role of miR-377 in AS via targeting Syk.
动脉粥样硬化（AS）是一种严重的疾病，其特征是大动脉中脂质和纤维的积累，是缺血性中风（IS）的最重要诱因。尽管已经在循环血液中发现了 microRNAs（miRNAs），但 miRNAs 在 AS 进展中的作用仍不清楚。在之前的研究中，我们证明了脾酪氨酸激酶（Syk）基因在 IS 过程中起着至关重要的作用。在本研究中，我们旨在阐明靶向 Syk 基因的 miRNAs 是否可能减缓 AS 的发展。通过生物信息学分析、RT-qPCR 和双荧光素酶报告基因检测，在 U937 和 THP-1 细胞中筛选候选 miRNAs。用载脂蛋白 E（ApoE）小鼠作为 AS 动物模型。通过 RAAV 转染鉴定 Syk 基因和 miRNAs 在 ApoE 小鼠 AS 发展中的作用。用 HE 染色、油红 O 染色和免疫组织化学染色确定 AS 的发生机制。用 RT-qPCR 和 Western blot 分别测定 miRNAs 和蛋白质的表达。Syk 基因的过表达加速了 AS 的发展。miR-377 在体外和体内实验中有效地介导了 Syk 基因的表达。进一步的分析表明，miR-377 通过下调 Syk 基因的表达部分缓解了 AS 的发展。本研究通过靶向 Syk 确定了 miR-377 在 AS 中的新作用。
