PMID: 26770639Jan 16, 2016Paper

Alteration of apoptotic protease activating factor 1 expression and possible role in ONOO(-)-induced apoptosis in human cerebral vascular smooth muscle cells

International Journal of Clinical and Experimental Medicine
Lin LiJianfeng Li

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...Continue Reading

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis