title: Agkistrodon acutus-purified protein C activator protects human umbilical vein endothelial cells against H2O2-induced apoptosis.
abstract: BACKGROUND
  Recent studies show that the Agkistrodon acutus (Viperidae) (syn. Deinagkistrodon acutus) protein C activator (PCA) treats acute myocardial infarction and ischaemia-reperfusion animal models effectively, while the underlying mechanism remains unknown.
  
  
  OBJECTIVE
  To study the effect of PCA on the injury of human umbilical vein endothelial cells (HUVECs) induced by H2O2and the underlying mechanism.
  
  
  METHODS
  Primary cultured HUVECs were pretreated with PCA (20, 40 and 80 μg/mL) for 1 h, then HUVEC apoptosis was induced by 300 μmol/mL H2O2. Apoptosis was analyzed by AnnexinV-FITC/PI, and reactive oxygen species (ROS) level was tested by flow cytometry. Colorimetric methods were used to detect the levels of NO and IL-1. In addition, real-time PCR and western blot analyses were used to detect the expression of eNOS and phospho-p38/MAPK.
  
  
  RESULTS
  Morphological changes were induced by H2O2in HUVECs. The cell survival rate was increased by 43.9, 64.0 and 80.6% in each PCA pretreated group (20, 40 and 80 μg/mL) compared to the model group. In each PCA pretreated group, oxidative stress level was also decreased to 54.7, 42.7 and 25.1%. Moreover, the level of IL-1 was decreased to 83.3, 62.2 and 30.7%. The level of NO was increased by 155.9, 232.4 and 317.6%. Apoptosis rate was decreased to 59.0, 47.7 and 32.7%. Phospho-p38 expression was downregulated, but eNOS expression was upregulated.
  
  
  CONCLUSIONS
  The results suggest that PCA can effectively protect the endothelial cells from injury induced by H2O2, which may be associated with antioxidation, upregulation of eNOS and downregulation of p38-MAPK.
authors: a Life Science College, Anhui Normal University , Wuhu , China.|b Department of Pathophysiology , Wannan Medical College , Wuhu , China.
authors: Shu
authors: S
authors: Li
authors: c Department of Ultrasonography , Yijishan Hospital , Wuhu , China.
authors: Yun
authors: Y
authors: Hong
authors: d Department of Basic Medicine , Wannan Medical College , Wuhu , China.
authors: Xin
authors: X
authors: Jin
authors: d Department of Basic Medicine , Wannan Medical College , Wuhu , China.
authors: Xianwei
authors: X
authors: Li
authors: d Department of Basic Medicine , Wannan Medical College , Wuhu , China.
authors: Entao
authors: E
authors: Sun
authors: b Department of Pathophysiology , Wannan Medical College , Wuhu , China.
authors: Genbao
authors: G
authors: Zhang
authors: d Department of Basic Medicine , Wannan Medical College , Wuhu , China.
authors: Linming
authors: L
authors: Lu
authors: a Life Science College, Anhui Normal University , Wuhu , China.
authors: Liuwang
authors: L
authors: Nie
mesh_terms: D017836:Agkistrodon; D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D011486:Protein C / Q000378:metabolism; D014757:Viper Venoms / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D011486:Protein C; D014757:Viper Venoms; D006861:Hydrogen Peroxide
keywords: HUVECs; PCA; ROS; eNOS; p38/MAPK