Anti-proliferative and anti-migratory effects of Scutellaria strigillosa Hemsley extracts against vascular smooth muscle cells
Anti-proliferative and anti-migratory effects of Scutellaria strigillosa Hemsley extracts against vascular smooth muscle cells
复制标题
黄芩提取物对血管平滑肌细胞的抗增殖和抗迁移作用
DOI：
10.1016/j.jep.2019.02.016
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发表时间：
2019-05-10
影响因子：
5.4
通讯作者：
Zhu, Jing
中科院分区：
文献类型：
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作者：
Li, Jiankun;Wang, Hairong;Zhu, Jing
民族药理学相关性：血管平滑肌细胞（VSMC）增殖和迁移的异常增加是包括再狭窄和动脉粥样硬化在内的心血管疾病（CVD）发病机制中的关键事件。临床和现代药理研究证实黄芩（中国通用名：黄芩）的干燥根对心血管疾病具有有益作用。黄芩中的黄酮类化合物具有抗增殖和抗迁移作用。与黄芩类似，黄芩（SSH）也被用于清热利湿，尤其富含汉黄芩素、汉黄芩苷、黄芩素和黄芩甙等黄酮类化合物。然而，关于SSH的药理活性报道较少。研究目的：研究黄芩Hemsley提取物(SSHE)的体内外抗增殖和抗迁移特性，并探讨其可能的作用机制。材料与方法：采用超高效液相色谱-三重飞行时间质谱法对SSHE的化学成分进行分析。 （UPLC-三重TOF-MS/MS）。分别使用 BrdU 掺入测定和细胞划痕试验研究细胞增殖和迁移。通过蛋白质印迹法测定蛋白质表达。在体内，我们建立了 C57BL/6 小鼠的动脉结扎模型，并口服 50 或 100 mg/kg/天的 SSHE。结扎后28天收获颈动脉并检查内膜中层厚度。结果：鉴定出12种化合物并初步表征。 SSHE显着抑制PDGF-BB刺激的VSMC增殖和迁移，并降低调节信号中间体的相对蛋白表达。此外，在 SSHE 预处理的 VSMC 中，SM22 α 的表达显着升高，而 SM22 α 的敲低则损害了 PDGF-BB 诱导的增殖和迁移停滞。同时，SSHE 预处理的 VSMC 中 ROS 生成和 ERK 磷酸化均减少。在颈动脉结扎小鼠模型中，SSHE处理显着抑制新生内膜增生。结论：SSHE显着抑制PDGF-BB诱导的VSMC增殖、迁移以及结扎引起的颈动脉新生内膜增生。 SM22 α 表达的上调、ROS 生成的抑制和 ERK 磷酸化至少是 SSHE 对 VSMC 影响的部分原因。
Ethnopharmacological relevance: The abnormal increase in vascular smooth muscle cell (VSMC) proliferation and migration are critical events in the pathogenesis of cardiovascular diseases (CVDs) including restenosis and atherosclerosis. The dried roots of Scutellaria baicalensis Georgi (common name: Huangqin in China) have been confirmed to possess beneficial effects on CVD by clinical and modern pharmacological studies. Flavonoids in Huangqin exert anti-proliferative and anti-migratory effects. Similar to Huangqin, Scutellaria strigillosa Hemsley (SSH) has been used to clear heat and damp and is especially rich in flavonoids including wogonin, wogonoside, baicalein, and baicalin. However, there have been few of reports about pharmacological activities of SSH.Aim of the study: To investigate the anti-proliferative and anti-migratory properties of Scutellaria strigillosa Hemsley extract (SSHE) in vitro and in vivo and explore its possible mechanism of action.Materials and methods: The chemical constituents of SSHE were analyzed by ultra-high performance liquid chromatography coupled with triple time-of-flight mass spectrometry (UPLC-Triple-TOF-MS/MS). Cell proliferation and migration were investigated using BrdU incorporation assay and cell scratch test, respectively. The protein expression was determined by western blotting. In vivo, we established an artery ligation model of C57BL/6 mice and orally administered them with 50 or 100 mg/kg/day of SSHE. The carotid arteries were harvested and the intima-media thickness was examined 28 days post-ligation.Results: Twelve compounds were identified and tentatively characterized. SSHE significantly inhibited the VSMC proliferation and migration stimulated by PDGF-BB and decreased the relative protein expression of regulatory signaling intermediates. Furthermore, the expression of SM22 alpha was significantly elevated in SSHE-pretreated VSMCs, whereas knockdown of SM22 alpha impaired the PDGF-BB-induced proliferation and migration arrest. Meanwhile, both ROS generation and the phosphorylation of ERK decreased in SSHE-pretreated VSMCs. In carotid artery ligation mice model, SSHE treatment significantly inhibited neointimal hyperplasia.Conclusions: SSHE significantly inhibited the PDGF-BB-induced VSMC proliferation, migration, and neointimal hyperplasia of carotid artery caused by ligation. Upregulation of SM22 alpha expression, inhibition of ROS generation and ERK phosphorylation were, at least, partly responsible for the effects of SSHE on VSMCs.