Department of Pharmacology, School of Chemical Biology & Pharmaceutical Sciences, Capital Medical University, Beijing 100069, China.
J Ethnopharmacol. 2009 Nov 12;126(2):197-206. doi: 10.1016/j.jep.2009.08.045. Epub 2009 Sep 6.
Danxiongfang (DF) is a new Chinese medicine formula used to treat atherosclerosis and vascular restenosis. The active ingredients in DF are danshensu (DSS), tanshinones (cryptotanshinone, CT) and ferulic acid (FA). The aim of present study was to evaluate the inhibitory effects of DF and its active ingredients on cell proliferation and protection against hydrogen peroxide (H(2)O(2))-induced injury in rat vascular smooth muscle cells (VSMC) in vitro.
VSMC proliferation was assayed by cell counting and measurement of cell viability using the 3-(4, 5-dimethylthiazol -2yl)-2, 5-diphenyltetrazolium bromide (MTT) method and protein content was measured by the Bradford method. The nitric oxide (NO) level was detected by an assay kit. The endothelin-1 (ET-1) level was measured by ELISA. The protective effects of DF and its active ingredients on H(2)O(2)-induced cell injury was evaluated in terms of cell viability (MTT assay), superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels. Hydroxyl free radicals generated by the Fenton reaction was detected with the spin-trapping technique on an electron spin resonance spectrometer.
The results suggest that DSS, CT, FA and DF inhibited VSMC proliferation by increasing the NO level and decreasing the ET-1 content. In rat VSMCs exposed to H(2)O(2), FA, DSS, CT and the six formulations of DF increased cell viability and SOD activity, and reduced the levels of MDA and hydroxyl free radicals. These effects of FA, DSS and CT occurred in a dose-dependent manner. Of the six formulas, DF 4 and DF 5 had the more significant activities. The effects of DF were much greater than those of the individual ingredients, even though the concentrations of these ingredients in the DF formulas were much lower than the doses of the individual ingredients used in each study, indicating markedly synergistic effects of DSS, CT and FA in DF on rat VSMCs.
these findings provide a pharmacological foundation for the clinical use of DF in the prevention and treatment of hyperlipidemia and atherosclerosis relevant to endothelial cell proliferation and damage.
丹芎方（DF）是一种用于治疗动脉粥样硬化和血管再狭窄的新中药配方。DF 的活性成分是丹参素（DSS）、丹参酮（隐丹参酮、CT）和阿魏酸（FA）。本研究旨在评估 DF 及其活性成分对体外大鼠血管平滑肌细胞（VSMC）增殖的抑制作用及对过氧化氢（H（2）O（2））诱导损伤的保护作用。
通过细胞计数和 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）法测量细胞活力来测定 VSMC 增殖，通过 Bradford 法测量蛋白质含量。通过试剂盒检测一氧化氮（NO）水平。通过 ELISA 法测量内皮素-1（ET-1）水平。通过 MTT 测定法、超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平评估 DF 及其活性成分对 H（2）O（2）诱导细胞损伤的保护作用。使用电子自旋共振谱仪上的自旋捕获技术检测 Fenton 反应产生的羟基自由基。
结果表明，DSS、CT、FA 和 DF 通过增加 NO 水平和降低 ET-1 含量来抑制 VSMC 增殖。在暴露于 H（2）O（2）的大鼠 VSMC 中，FA、DSS、CT 和 DF 的 6 种配方均增加了细胞活力和 SOD 活性，降低了 MDA 和羟基自由基水平。FA、DSS 和 CT 的这些作用呈剂量依赖性。在这 6 种配方中，DF 4 和 DF 5 的活性更强。DF 的作用比单独成分的作用大得多，尽管 DF 配方中的这些成分的浓度远低于每种研究中单独成分的剂量，这表明 DSS、CT 和 FA 在 DF 中对大鼠 VSMC 具有明显的协同作用。
这些发现为 DF 在预防和治疗与内皮细胞增殖和损伤相关的高脂血症和动脉粥样硬化中的临床应用提供了药理学基础。
