背景:动脉粥样硬化(AS)是一种常见的血管疾病,其主要影响因素是氧化低密度脂蛋白(ox-LDL)引起的内皮损伤。作为人参的主要活性成份之一,人参皂苷Rb3具有抗炎和抗氧化作用。然而,人参皂苷Rb3在ox-LDL诱导的内皮损伤中的作用尚不清楚。
目的:本研究旨在评估人参皂苷Rb3对ox-LDL处理的人主动脉内皮细胞(HAECs)的作用和潜在机制。
方法:用ox-LDL处理的HAECs建立体外AS模型。用细胞计数试剂盒-8(CCK-8)分析HAEC的活力。进行流式细胞术以评估细胞凋亡。氧化应激,使用酶联免疫吸附测定(ELISA)和蛋白质印迹法评估炎症和内皮功能障碍。使用定量实时聚合酶链反应(qPCR)评估miR-513a-5p的水平。进行双荧光素酶测定以分析miR-513a-5p与锌指和含BTB结构域的蛋白(ZBTB20)之间的关系。
结果:HAECs暴露于ox-LDL(50μg/mL)会降低细胞活力,超氧化物歧化酶(SOD)活性和内皮型一氧化氮合酶(eNOS)表达,在增加丙二醛(MDA)水平的同时,白细胞介素6(IL-6),肿瘤坏死因子α(TNF-α),和可溶性细胞间粘附分子-1(sICAM-1)。用Rb3预处理显着增强细胞活力并降低ox-LDL诱导的氧化应激,HAECs的炎症和内皮功能障碍。ox-LDL降低miR-513a-5p水平,通过Rb3预处理逆转。ZBTB20是HAECs中miR-513a-5p的靶标,和ox-LDL上调ZBTB20表达,通过Rb3预处理逆转。miR-513a-5p抑制Rb3对ox-LDL诱导的HAECs的保护作用减弱,被ZBTB20敲低逆转。
结论:人参皂苷Rb3通过调节miR-513a-5p/ZBTB20轴降低ox-LDL对HAECs的影响,为AS的治疗提供了理论依据。
BACKGROUND: Atherosclerosis (AS) is a common vascular disease, and its main influencing factor is endothelial damage caused by oxidized low-density lipoprotein (ox-LDL). As one of the main active ingredients of ginseng, ginsenoside Rb3 has anti-inflammatory and anti-oxidative effects. However, the role of ginsenoside Rb3 in endothelial injury induced by ox-LDL is not clear.
OBJECTIVE: This study aimed to evaluate the effect and potential mechanism of ginsenoside Rb3 action on ox-LDL-treated human aortic endothelial cells (HAECs).
METHODS: The HAECs treated with ox-LDL were used to establish an in vitro AS model. The viability of the HAECs was analyzed with Cell Counting Kit-8 (CCK-8). Flow cytometry was performed to assess the apoptosis. Oxidative stress, inflammation and endothelial dysfunction were evaluated using enzyme-linked immunosorbent assay (ELISA) and western blotting. The levels of miR-513a-5p were assessed using quantitative real-time polymerase chain reaction (qPCR). A dual-luciferase assay was performed to analyze the relationship between miR-513a-5p and a zinc finger and BTB domain-containing protein (
ZBTB20).
RESULTS: Exposure of HAECs to ox-LDL (50 μg/mL) reduced cell viability, superoxide dismutase (SOD) activity and endothelial nitric oxide synthase (eNOS) expression, while increasing the levels of malondialdehyde (MDA), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), and soluble intercellular adhesion molecule-1 (sICAM-1). The pretreatment with Rb3 markedly enhanced cell viability and decreased ox-LDL-induced oxidative stress, inflammation and endothelial dysfunction in HAECs. The ox-LDL decreased the level of miR-513a-5p, which was reversed by Rb3 pretreatment. The
ZBTB20 was a target of miR-513a-5p in HAECs, and ox-LDL upregulated
ZBTB20 expression, which was reversed by Rb3 pretreatment. The protective effect of Rb3 on ox-LDL-induced HAECs was diminished by miR-513a-5p inhibition, which was reversed by
ZBTB20 knockdown.
CONCLUSIONS: Ginsenoside Rb3 reduces the effects of ox-LDL on HAECs by regulating the miR-513a-5p/
ZBTB20 axis, which provides a theoretical basis for the treatment of AS.