Zhang Yu-yan, Zhou Hui-fen, Yang Jie-hong, He Yu, Chen Xiao-qiang, Nishinari Katsuyoshi, Wan Hao-fang, Wan Hai-tong
Research Institution of Cardio-Cerebral-Vascular Disease, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
Pharmaceutical College, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
Chin J Integr Med. 2016 Apr;22(4):276-83. doi: 10.1007/s11655-016-2498-x. Epub 2016 Apr 9.
To observe the effects of Danhong Injection (丹红注射液) and its main components, including daiclzein and hydroxysafflor yellow A (HSYA), on the anticoagulation, fibrinolysis, anti-apoptosis in hypoxia model of vein endothelial cells (VECs).
VECs were prepared and were put in a hypoxia environment, which consisted of mixed gas of 95% N and 5% CO mixed gas, when reached confluent culture. Five groups used different treatments, including normal control group, hypoxia group, daiclzein group, HSYA group and Danhong Injection group. The VECs were identified by fluorescence double labeling methods. The morphology was observed by a phase contrast microscopy. The effects of Danhong Injection, daiclzein and HSYA on 6 keto prostaglandin F1α (6-keto-PGF1α) level was measured by the method of radioimmunoassay (RIA). Superoxide dismutase (SOD) activity was tested by water soluble tetrazolium salt. The content of malondialdehyde (MDA) was measured by thiobarbituric acid. The activities of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) were measured by the method of chromogenic substrate. The contents of endothelin (ET) and nitric oxide (NO) were detected by non-equilibrium RIA and enzymelinked immunosorbent assay. Cells apoptosis rate was determined by flow cytometry.
Compared with the normal control group, the floating cells number, PAI activity, ET and MDA contents, and cells apoptosis rate in the culture solution of hypoxia group were all significantly increased, whereas the 6-keto-PGF1α and NO contents, and t-PA and SOD activities were decreased significantly (P<0.01). Compared with the hypoxia group, Danhong Injection markedly increased the 6-keto-PGF1α content and SOD activity, regulated PAI and t-PA activities, ET and NO contents, and decreased MDA content and cells apoptosis rate (P<0.05 or P<0.01).
Danhong Injection and its main components played an important role in protecting primary VECs from hypoxic damage by regulating the secretion and vasomotor function of VECs. The function of Danhong Injection was most remarkable.
观察丹红注射液及其主要成分大豆苷元、羟基红花黄色素A（HSYA）对静脉内皮细胞（VECs）缺氧模型抗凝、纤溶及抗凋亡的影响。
制备VECs，待其达到融合培养后置于由95%N和5%CO混合气体组成的缺氧环境中。五组采用不同处理，包括正常对照组、缺氧组、大豆苷元组、HSYA组和丹红注射液组。采用荧光双标记法鉴定VECs。用相差显微镜观察细胞形态。采用放射免疫分析法（RIA）检测丹红注射液、大豆苷元及HSYA对6-酮前列腺素F1α（6-keto-PGF1α）水平的影响。采用水溶性四氮唑盐检测超氧化物歧化酶（SOD）活性。采用硫代巴比妥酸检测丙二醛（MDA）含量。采用发色底物法检测组织型纤溶酶原激活剂（t-PA）和纤溶酶原激活剂抑制剂（PAI）活性。采用非平衡RIA和酶联免疫吸附测定法检测内皮素（ET）和一氧化氮（NO）含量。采用流式细胞术测定细胞凋亡率。
与正常对照组相比，缺氧组培养液中漂浮细胞数、PAI活性、ET和MDA含量以及细胞凋亡率均显著升高，而6-keto-PGF1α和NO含量以及t-PA和SOD活性显著降低（P<0.01）。与缺氧组相比，丹红注射液显著提高了6-keto-PGF1α含量和SOD活性，调节了PAI和t-PA活性、ET和NO含量，降低了MDA含量和细胞凋亡率（P<0.05或P<0.01）。
丹红注射液及其主要成分通过调节VECs的分泌和血管舒缩功能，在保护原代VECs免受缺氧损伤方面发挥重要作用。丹红注射液的作用最为显著。
