title: Effects of Danhong Injection (丹红注射液) and its main components on anticoagulation and fibrinolysis in cultured vein endothelial cells.
abstract: OBJECTIVE
  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).
  
  
  METHODS
  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.
  
  
  RESULTS
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Research Institution of Cardio-Cerebral-Vascular Disease, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
authors.forename: Yu-yan
authors.initials: YY
authors.lastname: Zhang
authors.affiliation: Research Institution of Cardio-Cerebral-Vascular Disease, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
authors.forename: Hui-fen
authors.initials: HF
authors.lastname: Zhou
authors.affiliation: Research Institution of Cardio-Cerebral-Vascular Disease, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
authors.forename: Jie-hong
authors.initials: JH
authors.lastname: Yang
authors.affiliation: Pharmaceutical College, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: He
authors.affiliation: School of Food and Pharmaceutical Engineering, Hubei University of Technology, Wuhan, 430068, China.
authors.forename: Xiao-qiang
authors.initials: XQ
authors.lastname: Chen
authors.affiliation: School of Food and Pharmaceutical Engineering, Hubei University of Technology, Wuhan, 430068, China.
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Nishinari
authors.affiliation: Pharmaceutical College, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China.
authors.forename: Hao-fang
authors.initials: HF
authors.lastname: Wan
authors.affiliation: Research Institution of Cardio-Cerebral-Vascular Disease, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China. whtong@126.com.
authors.forename: Hai-tong
authors.initials: HT
authors.lastname: Wan
mesh_terms: D015121:6-Ketoprostaglandin F1 alpha / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001777:Blood Coagulation / Q000187:drug effects*; D002452:Cell Count; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D016232:Endothelins / Q000378:metabolism; D005169:Factor VIII / Q000378:metabolism; D005342:Fibrinolysis / Q000187:drug effects*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007231:Infant, Newborn; D007267:Injections; D008315:Malondialdehyde / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D015849:Plasminogen Inactivators / Q000378:metabolism; D011817:Rabbits; D013482:Superoxide Dismutase / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology*
chemical_list: D004365:Drugs, Chinese Herbal; D016232:Endothelins; D015849:Plasminogen Inactivators; C540581:danhong; D009569:Nitric Oxide; D008315:Malondialdehyde; D015121:6-Ketoprostaglandin F1 alpha; D005169:Factor VIII; D013482:Superoxide Dismutase; D010959:Tissue Plasminogen Activator
keywords: Chinese medicine; Danhong Injection; anticoagulation; daiclzein; fibrinolysis; hydroxysafflor yellow A; vein endothelial cells