Nowak D, Bialasiewicz P, Piasecka G, Antczak A, Krol M
Department of Pneumology and Allergology, Medical University of Lodz, Poland.
Int J Clin Lab Res. 1997;27(3):189-94. doi: 10.1007/BF02912456.
Polymorphonuclear leukocytes isolated from peripheral blood of asthmatics appear to be primed to release more reactive oxygen species than cells of healthy subjects. The enhanced agonist-induced rise in the intracellular free calcium concentration may be responsible for this increased respiratory burst. To test this hypothesis we studied the N-formyl-methionyl-leucyl-phenylalanine- and cyclopiazonic acid--(an inhibitor of Ca(2+)-ATPase of intracellular calcium stores) induced calcium increase in the polymorphonuclear leukocytes of 28 subjects (16 with moderate asthma, 69.6% +/- 8.3% predicted normal peak expiratory flow and 12 normal controls) using a fluorescent probe Fura-2AM at 100 nM and 1 mM extracellular calcium concentrations. In 1 mN calcium, the N-formyl-methionyl-leucyl-phenylalanine-induced calcium increase was 1.7-fold higher in asthmatics than in healthy subjects. Similarly, the contribution of calcium from intracellular stores to the calcium response to N-formyl-methionyl-leucyl-phenylalanine was higher in asthmatics (55% +/- 14% vs. 39% +/- 14%, P < 0.01). The pool of calcium released from intracellular stores by N-formyl-methinoyl-leucyl-phenylalanine and cyclopiazonic acid was 2.3- and 2.2-fold larger than in control cells. There was a correlation between maximal intracellular calcium concentration related to N-formyl-methionyl-leucyl-phenylalanine-induced calcium release from intracellular stores and forced expiratory volume in 1 s expressed as percentage predicted and reversibility in asthmatics (r = 0.63, r = -0.53, P < 0.05). In conclusion, polymorphonuclear leukocytes of asthmatics exhibit an altered calcium response that is mainly dependent on increased calcium release from intracellular stores.
从哮喘患者外周血中分离出的多形核白细胞似乎比健康受试者的细胞更易于释放更多的活性氧。细胞内游离钙浓度因激动剂诱导而升高，这可能是呼吸爆发增加的原因。为了验证这一假设，我们使用荧光探针Fura - 2AM，在细胞外钙浓度为100 nM和1 mM的条件下，研究了28名受试者（16名中度哮喘患者，预计正常呼气峰值流量为69.6%±8.3%，以及12名正常对照者）的多形核白细胞中，N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸和环匹阿尼酸（一种细胞内钙储存的Ca(2 +)-ATP酶抑制剂）诱导的钙增加情况。在1 mM钙浓度下，哮喘患者中N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸诱导的钙增加比健康受试者高1.7倍。同样，哮喘患者中细胞内钙储存对N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸钙反应的贡献更高（55%±14%对39%±14%，P < 0.01）。N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸和环匹阿尼酸从细胞内储存释放的钙池比对照细胞大2.3倍和2.2倍。在哮喘患者中，与N - 甲酰 - 甲硫氨酰 - 亮氨酰 - 苯丙氨酸诱导的细胞内钙储存释放相关的最大细胞内钙浓度与以预计百分比表示的1秒用力呼气量和可逆性之间存在相关性（r = 0.63，r = -0.53，P < 0.05）。总之，哮喘患者的多形核白细胞表现出钙反应改变，这主要取决于细胞内储存释放钙的增加。
