title: [Effect of C5a on expression of thrombomodulin in endothelial cells in vitro].
abstract: OBJECTIVE
  To investigate the effect of C5a on the expression of thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs).
  
  
  METHODS
  HUVECs cultured in vitro were stimulated with C5a for 8, 12, 16, 20 hours in a concentration of 200 microg/L, and also with different concentrations of 100, 200, 300 microg/L for 12 hours respectively. The levels of both mRNA and protein expression of TM was detected by real-time polymerase chain reaction (PCR) and Western blotting respectively, and the dose and time dependent effects of C5a on the expression of TM were evaluated.
  
  
  RESULTS
  C5a down-regulated the TM expression at both protein and mRNA level. The down-regulation was time-dependent [(93.11+/- 1.57) x10(-2) , (71.05+/-3.39)x10(-2), (65.48+/-4.28)x10(-2), (62.69+/-4.03)x10(-2)at protein level and (301.71+/-80.40)x10(-6), (38.29+/-20.24)x10(-6), (8.82+/-2.66)x10(-6), (7.05+/-0.80)x10(-6) at mRNA level, all P<0.05] and dose-dependent [(113.25+/-3.97)x10(-2), (80.18+/-2.56)x10(-2), (73.22+/- 4.36)x10(-2) at protein level and (401.77+/-20.46)x10(-6), (31.12+/-3.51)x10(-6), (18.19+/-1.46)x10(-6) at mRNA level , all P<0.05]. When concentration of C5a at 300 microg/L was used to stimulate HUVECs for longer than 12 hours, the lowering of TM at protein level was slowed down obviously. And concentration of C5a at 200 microg/L was used to stimulate HUVECs for 12 hours, the lowering of TM at mRNA level was slowed down obviously.
  
  
  CONCLUSIONS
  C5a can depress the gene expression of TM, and then affect the protein's translation. By this means, C5a can lead to hypercoagulability and inflammatory injuries in sepsis.
authors.affiliation: Guangzhou Medical College, Guangzhou 510010, Guangdong, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Fang
authors.forename: Zhen-hui
authors.initials: ZH
authors.lastname: Guo
authors.forename: Bao-quan
authors.initials: BQ
authors.lastname: Zhang
authors.forename: Xiao-feng
authors.initials: XF
authors.lastname: Wu
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Feng-lin
authors.initials: FL
authors.lastname: Lv
authors.forename: Lei
authors.initials: L
authors.lastname: Su
mesh_terms: D002478:Cells, Cultured; D015936:Complement C5a / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D018180:Thrombomodulin / Q000235:genetics / Q000378:metabolism*; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D012333:RNA, Messenger; D018180:Thrombomodulin; D015936:Complement C5a