title: Effects of different sustained hydrostatic pressures on connexin 43 in human bladder smooth muscle cells.
abstract: OBJECTIVE
  To cultivate primary human bladder smooth muscle cells (hBSMCs) in vitro and to study the influences of different sustained hydrostatic pressures upon hBSMCs in terms of cell proliferation and expression of gap-junction connexin 43 (Cx43).
  
  
  METHODS
  hBSMCs were exposed to 0, 20, 40, 60, 80 and 100 cm H(2)O sustained hydrostatic pressures for 6, 12, 24 and 72 h, respectively. hBSMC proliferation was quantified with Cell Counting Kit-8 (CCK-8) afterwards. Immunoﬂuorescence staining and Western blotting were employed to determine hBSMC Cx43 expression.
  
  
  RESULTS
  hBSMC proliferation increased with time (6-72 h) without significant differences between pressure levels at each time point. Expression of Cx43 on Western blotting and immunoﬂuorescence staining was similar under varied pressures for 6 and 12 h while seeing significant decreases under hydrostatic pressures >60 cm H(2)O for 24 h or pressures >40 cm H(2)O for 72 h.
  
  
  CONCLUSIONS
  Different sustained hydrostatic pressures (<100 cm H(2)O) bear no significant effects on proliferation of hBSMCs. Interestingly, hydrostatic pressures above the physiological level seem to cause the gap junctions of hBSMCs to decrease significantly and affect the cellular functions in the cultivation group over a longer period of time.
authors.affiliation: Department of Urology, West China Hospital/West China School of Medicine, Sichuan University, Chengdu, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Kunjie
authors.initials: K
authors.lastname: Wang
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
mesh_terms: D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D006874:Hydrostatic Pressure; D040542:Mechanotransduction, Cellular; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D013997:Time Factors; D001743:Urinary Bladder / Q000166:cytology / Q000378:metabolism*
chemical_list: D018031:Connexin 43; C500216:GJA1 protein, human