科研之友
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摘要
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. Apaf1 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.
