title: Alteration of apoptotic protease activating factor 1 expression and possible role in ONOO(-)-induced apoptosis in human cerebral vascular smooth muscle cells.
abstract: The study was to investigate the change of apoptotic protease activating factor 1 (Apaf1) expression in the human cerebral vascular smooth muscle cells (HCVSMCs) in response to peroxynitrite (ONOO(-)). HCVSMCs were cultured in vitro and ONOO(-) of different concentrations was directly added to the culture medium. The total proteins were extracted and the alteration of Apaf1 expression was examined by the means of Western-blot. Apaf-1 siRNA (h) was put into another plate of HCVSMCs for transfection. The transfected cells were also incubated in different concentrations of ONOO(-). The change of Apaf1 protein expression after siRNA transfection was examined by the means of Western-blot. The morphological changes were observed by acridine orange staining to determine whether cells experienced apoptosis in response to ONOO(-) before and after siRNA. The Flow cytometry analysis was used to examine the change of cells apoptotic rates in response to ONOO(-) before and after siRNA. Obviously, there was up-regulated Apaf1 expression at protein level in the course of HCVSMCs apoptosis induced by ONOO(-). When Apaf1 expression was suppressed, the apoptotic sum of HCVSMCs didn't change. This study demonstrates that Apaf1 gene is involved in ONOO(-)-induced apoptosis in HCVSMCs. Whether HCVSMCs treated by ONOO(-) undergo apoptosis depends on Apaf1 level.
authors.affiliation: Binzhou Medical UniversityYantai, Shandong, P. R. China; Cancer Research Center, Shandong UniversityJinan, Shandong, P. R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: Binzhou Medical University Yantai, Shandong, P. R. China.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zhao
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Provincial Hospital Affiliated to Shandong University Jinan, Shandong, P. R. China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Li
keywords: ONOO-; apoptosis; apoptotic protease activating factor 1; human cerebral vascular smooth muscle cell