DeMeester S L, Buchman T G, Qiu Y, Jacob A K, Dunnigan K, Hotchkiss R S, Karl I, Cobb J P
Department of Surgery, Washington University School of Medicine, St Louis, Mo, USA.
Arch Surg. 1997 Dec;132(12):1283-7; discussion 1287-8. doi: 10.1001/archsurg.1997.01430360029005.
To determine whether prior heat shock would attenuate endothelial cell apoptosis and whether any effect of preemptive heat shock is mediated through a nuclear factor kappa B and inhibitor kappa B alpha mechanism.
A randomized, controlled in vitro study.
A laboratory in a large, academic medical center.
Cultured primary porcine endothelial cells were treated with increasing doses of sodium arsenite (40-160 micromol/L), after which the interval until subsequent apoptotic (lipopolysaccharide-arsenite) challenge was varied (4-16 hours). The degree of cell death and apoptosis were determined using neutral red uptake and staining with annexin V and propidium iodide, respectively. Inducible heat shock protein 70 and inhibitor kappa B alpha levels in treated cells were determined by Western blot analysis. Lipopolysaccharide-induced nuclear factor kappa B activity was assessed using an electrophoretic mobility shift assay.
Prior arsenite treatment decreased cell death by apoptosis in a time- and dose-dependent manner. Specifically, a higher sodium arsenite concentration and shorter intervals afforded better protection (P=.01, 160 micromol/L at 4 hours). Protection against apoptosis correlated with increased heat shock protein 70 and inhibitor kappa B alpha levels and decreased nuclear factor kappa B binding activity.
Arsenite, an inducer of the heat shock response, decreased stress-induced endothelial cell apoptosis. The mechanism of this protection may include decreased nuclear factor kappa B activity or increased inducible heat shock protein 70 levels. Heat shock protein 70 may serve as a molecular marker to determine not only the phenotypic state of the cell but also the durability of protection afforded by heat shock. These data support the hypothesis that stress-induced changes in transcription factor activity and protein expression can regulate the induction of apoptosis.
确定预先热休克是否会减轻内皮细胞凋亡，以及预先热休克的任何效应是否通过核因子κB和抑制因子κBα机制介导。
一项随机对照体外研究。
一家大型学术医学中心的实验室。
用递增剂量的亚砷酸钠（40 - 160 μmol/L）处理培养的原代猪内皮细胞，之后改变直至随后凋亡（脂多糖 - 亚砷酸盐）刺激的间隔时间（4 - 16小时）。分别使用中性红摄取以及用膜联蛋白V和碘化丙啶染色来确定细胞死亡和凋亡程度。通过蛋白质印迹分析测定处理细胞中诱导型热休克蛋白70和抑制因子κBα水平。使用电泳迁移率变动分析评估脂多糖诱导的核因子κB活性。
预先亚砷酸盐处理以时间和剂量依赖方式减少凋亡引起的细胞死亡。具体而言，较高的亚砷酸钠浓度和较短的间隔时间提供了更好的保护（P = 0.01，4小时时为160 μmol/L）。对凋亡的保护与热休克蛋白70和抑制因子κBα水平增加以及核因子κB结合活性降低相关。
亚砷酸盐作为热休克反应的诱导剂，减少应激诱导的内皮细胞凋亡。这种保护机制可能包括核因子κB活性降低或诱导型热休克蛋白70水平增加。热休克蛋白70不仅可作为确定细胞表型状态的分子标志物，还可作为确定热休克提供的保护持续性的分子标志物。这些数据支持应激诱导的转录因子活性和蛋白质表达变化可调节凋亡诱导的假说。
