title: Reduced proliferation of aged human vascular smooth muscle cells--role of oxygen-derived free radicals and BubR1 expression.
abstract: BACKGROUND
  Aging is a risk factor for atherosclerosis. Recent studies suggest cell cycle events as well as reactive oxygen species (ROS) contribute to vascular cell dysfunction associated with aging. Mice expressing low levels of the spindle assembly checkpoint protein BubR1 develop aging-associated vascular changes at a young age, including decreased smooth muscle cells and increased reactive oxygen species (ROS) production. This study was designed to determine the effect of aging and production of oxygen-derived free radicals on expression of BubR1.
  
  
  METHODS
  To assess cell proliferation capacity, human aortic smooth muscle cells (hAoSMC) derived from a young group (17-30 y) or an aged group (57-62 y) were cultured, and cell numbers were directly counted in using a Neubauer chamber. RT-PCR assay was used to evaluate BubR1 expression in cultured hAoSMC stimulated with Angiotensin II or H(2)O(2).
  
  
  RESULTS
  No significant difference in BubR1 expression or hAoSMC proliferative ability was demonstrated at passage 5, but both were significantly decreased at passage 8 in the aged hAoSMC. Angiotensin II and H(2)O(2) up-regulated BubR1 expression in young hAoSMC, and the up-regulation was abrogated by a p38 MAPK inhibitor or an inhibitor of the NADH/NADPH oxidase. siRNA against BubR1 reduced proliferative activity and increased ROS production in hAoSMC.
  
  
  CONCLUSIONS
  These findings demonstrate BubR1 mRNA expression decreases along with proliferation in aged hAoSMC. Aging-related loss of BubR1 and subsequent impairment of reactivity to ROS may explain reduced proliferative capacity of aged smooth muscle cells.
authors.affiliation: Department of Surgery, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Guntani
authors.forename: Takuya
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Ryoichi
authors.initials: R
authors.lastname: Kyuragi
authors.forename: Kazuomi
authors.initials: K
authors.lastname: Iwasa
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Onohara
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Itoh
authors.forename: Zvonimir S
authors.initials: ZS
authors.lastname: Katusic
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Maehara
mesh_terms: D000293:Adolescent; D000328:Adult; D000375:Aging / Q000378:metabolism / Q000473:pathology*; D000804:Angiotensin II / Q000494:pharmacology; D018797:Cell Cycle Proteins; D049109:Cell Proliferation; D002478:Cells, Cultured; D005609:Free Radicals; D005786:Gene Expression Regulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008875:Middle Aged; D009131:Muscle, Smooth, Vascular / Q000473:pathology*; D032389:Myocytes, Smooth Muscle / Q000473:pathology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: C584278:Bub1b protein, mouse; D018797:Cell Cycle Proteins; D005609:Free Radicals; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D000804:Angiotensin II; D006861:Hydrogen Peroxide; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases