title: [An ischemic postconditioning model of human dermal microvascular endothelial cells].
abstract: OBJECTIVE
  To establish a model of hypoxic postconditioning of dermal microvascular endothelial cells.
  
  
  METHODS
  During reoxygenation after hypoxia, cells received three times of hypoxic/ reoxygenation alternate treatment for a certain time. The cells were seeded on 6-well plates, with one plate for one group. They were divided into 6 groups as group 1 ( Control), group 2 (8h hypoxia + 24h reoxygenation), group 3 (8h hypoxia + 2 min x 3 times post-hypoxia treatment) , group 4 (8h hypoxia + 5 min x 3 times post-hypoxia treatment), group 5 (8h hypoxia + 10 min x 3 times post-hypoxia treatment), 6 group (8h hypoxia + 20 min x 3 times post-hypoxia treatment). Each group underwent 8 h hypoxia + 24 h hypoxia Buffer and reoxygenation. Lactate dehydrogenase (LDH) was detected during the process. Apoptosis rate was calculated by staining Tunel method. Bcl-2, Bax and activated caspase-3 protein were detected by Western Blot.
  
  
  RESULTS
  In the continuous hypoxia process, the LDH was (1563 ± 83.35) IU/L at 8h and (582.85 ± 58.25 ) IU/L at 0h, showing a statistical difference (P = 0.0001). Western blotting results showed that the expression of Bax/Bcl-2 in group 2 was 0.38 ± 0.02, showing a significant difference when compared with that in group 3 (0.23 ± 0.01) and group 4 (0.22 ± 0.02) (P = 0.012, P = 0.005), while not when compared with that in group 5 (0.33 ± 0.02) and 6 groups (0.34 ± 0.01) P > 0.05). The ratio of Activated caspase-3/caspase-3 in group 2 (6.30 ± 1.50) was significantly higher than that in group 3 (2.17 ± 0.26) and group 4 (2.63 ± 0.31) (P = 0.008, P = 0.019); while not in group 5 (4.36 ± 0.29) and group 6 (4.97 ± 0.51) (P > 0.05).
  
  
  CONCLUSIONS
  The model of hypoxic postconditioning of human dermal microvascular endothelial cells is successfully established.
authors.forename: Lu
authors.initials: L
authors.lastname: Kun
authors.forename: Chu
authors.initials: C
authors.lastname: Xiaoling
authors.forename: Wang
authors.initials: W
authors.lastname: Lyuya
authors.forename: Lin
authors.initials: L
authors.lastname: Huang
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D053148:Caspase 3 / Q000032:analysis; D015687:Cell Hypoxia; D042783:Endothelial Cells / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D057775:Ischemic Postconditioning / Q000379:methods*; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D010101:Oxygen Consumption; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000032:analysis
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3