Jobin C, Gauthier J
Department of Medicine, University of North Carolina at Chapel Hill 27599, USA.
Inflammation. 1997 Apr;21(2):235-50. doi: 10.1023/a:1027326405788.
We have analyzed the effect of cellular density of 5-Lipoxygenase (5-LO), 5-lipoxygenase-activating protein (FLAP) and interleukin-1 beta (IL-1 beta) gene expression in neutrophils from healthy subjects under culture conditions of low and high cell density. By using RT-PCR techniques, we have found that 5-LO mRNA accumulation decreased in cells cultured at high density, while FLAP mRNA is not affected. De novo 5-LO synthesis, as well as steady-state levels, were reduced in cells maintained at high density. In contrast, the high density conditions lead to the induction of IL-1 beta gene at the RNA and protein levels as measured by RT-PCR and by immunoprecipitation. These results suggest that cellular density plays a role in gene modulation when neutrophils are accumulating at an inflammatory site since neutrophils obtained from the synovial fluid of patients with RA exhibit a protein synthesis profile similar to that observed in peripheral blood neutrophils cultured at high density.
我们分析了在低细胞密度和高细胞密度培养条件下，健康受试者中性粒细胞中5-脂氧合酶（5-LO）、5-脂氧合酶激活蛋白（FLAP）和白细胞介素-1β（IL-1β）基因表达的细胞密度效应。通过使用逆转录聚合酶链反应（RT-PCR）技术，我们发现高密度培养的细胞中5-LO信使核糖核酸（mRNA）积累减少，而FLAP mRNA不受影响。高密度培养的细胞中，5-LO的从头合成以及稳态水平均降低。相反，通过RT-PCR和免疫沉淀法测定，高密度条件导致RNA和蛋白质水平上IL-1β基因的诱导。这些结果表明，当中性粒细胞在炎症部位积聚时，细胞密度在基因调节中起作用，因为从类风湿性关节炎（RA）患者滑液中获得的中性粒细胞表现出与高密度培养的外周血中性粒细胞中观察到的蛋白质合成谱相似。
