title: Shenfu injection protects human ECV304 cells from hydrogen peroxide via its anti-apoptosis way.
abstract: BACKGROUND
  The pathogenesis of thromboangiitis obliterans (TAO) has not been fully elucidated until now. Shenfu injection (SFI), a traditional Chinese formula has been widely used clinically for the treatment of cardiovascular diseases for more than two decade. Our previous results first suggested that SFI can cause a significant therapeutic effect on experimental TAO model rats. This experiment was designed to further investigate the protective effect of SFI on VEC damaged by hydrogen peroxide (H2O2) oxidative stress in vitro.
  The cell viability was evaluated by the MTT assay, the activities of SOD and GSH-PX and the content of MDA in the supernatants of the cultured ECV304 cells were evaluated by a colorimetry method, cell apoptosis was detected by flow cytometry and an AO/EB double staining method. The protein expressions of Bcl2, Bax and caspase-3 were examined by Western blotting.
  
  
  RESULTS
  When compared with control group, lower survival rate of ECV304 cells was observed in H2O2 group (p<0.01) ; 20μl/ml, 30μl/ml and 40μl/ml SFI increased the survival rate of ECV304 cells under H2O2 oxidative stress (p<0.05 and p<0.01). The activities of SOD and GSH-PX were higher and MDA level was lower in H2O2 group than those in control group. These effects of H2O2 on SOD, GSH-PX activities and MDA content were reversed by SFI in concentration-dependent way (p<0.05 and p<0.01). Flow cytometry and AO-EB double staining discovered that SFI pretreatment inhibited the ECV304 cells apoptosis. The protein expression of caspase3 in 30μl/ml and 40μl/ml SFI groups significantly decreased whereas Bcl2 protein expressions in 20μl/ml, 30μl/ml and 40μl/ml SFI groups were higher than H2O2 group, with Bax protein expression much lower than H2O2 group (p<0.05 and p<0.01).
  
  
  CONCLUSIONS
  Our findings suggest that SFI could prevent the ECV304 cells against H2O2 oxidative-stress by enhancing antioxidant enzyme activities, reducing the membrane lipid peroxidation, as well as upregulating antiapoptotic and downregulating apoptosis protein expressions.
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China; Department of Experimental Teaching, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Hong
authors: H
authors: Fen-fang
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Guo
authors: G
authors: Fa-xian
authors: Department of Tissue Embryology of Basic Medical College, Nanchang 310006, China.
authors: Zhou
authors: Z
authors: Ying
authors: The First Affiliated Hospital of Obstetrics and Gynecology, Nanchang 310006, China.
authors: Min
authors: M
authors: Qin-hua
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Zhang
authors: Z
authors: Da-lei
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Yang
authors: Y
authors: Bei
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Zhou
authors: Z
authors: Wei-ying
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Wu
authors: W
authors: Lei
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Wei
authors: W
authors: Zhi-ping
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China.
authors: Liu
authors: L
authors: Hui
authors: Department of Physiology, College of Medicine, Nanchang University, Nanchang 310006, China. Electronic address: slyang@ncu.edu.cn.
authors: Yang
authors: Y
authors: Shu-long
mesh_terms: D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D004365:Drugs, Chinese Herbal; D006801:Humans; D006861:Hydrogen Peroxide; D007267:Injections; D018384:Oxidative Stress / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000975:Antioxidants; C494826:BAX protein, human; D004365:Drugs, Chinese Herbal; D019253:Proto-Oncogene Proteins c-bcl-2; C492793:Shen-Fu; D051028:bcl-2-Associated X Protein; D006861:Hydrogen Peroxide; D053148:Caspase 3
keywords: Anti-oxidation; Apoptosis; H(2)O(2); Shenfu injection; Vascular endothelial cell