新型PDE3抑制剂西洛他唑对fMLP诱导的HL-60细胞超氧自由基生成和钙离子浓度升高的影响

Effects of a novel PDE3 inhibitor, cilostazol on fMLP-induced superoxide production and calcium concentration increase in HL-60 cells

目的 研究新型磷酸二酯酶 (PDE) 3抑制剂西洛他唑(cilostazol)对中性粒细胞超氧自由基 (O·2 )和钙离子浓度([Ca2 + ]i)的影响。方法 体外培养急性早幼粒细胞白血病HL 6 0细胞 ,以 1 3%DMSO分化诱导为中性粒细胞样细胞后 ,采用特制新型仪器 ,同时实现化学发光法测定O·2 和荧光法测定 [Ca2 + ]i,观察西洛他唑对趋化肽N 甲酰 甲硫氨酰 亮氨酰苯丙氨酸 (fMLP)刺激引起的中性粒细胞样HL 6 0细胞O·2 生成和 [Ca2 + ]i 升高的影响以及二者变化的时相关系。结果 西洛他唑 (1～ 30 μmol·L-1)明显抑制由fMLP诱导的中性粒细胞O·2 生成和 [Ca2 + ]i 升高 ,其中 10和 30 μmol·L-1浓度的西洛他唑组与对照组相比差异有统计学意义 (P<0 0 1)。在时相关系上 ,fMLP诱导O·2 生成的达峰时间较之 [Ca2 + ]i 的变化略显滞后 ,而 10 μmol·L-1西洛他唑对fMLP诱导O·2 生成和 [Ca2 + ]i 升高的影响表现出一致性 ,几乎同步。结论 西洛他唑对fMLP 诱导的中性粒细胞样细胞内O·2 生成和 [Ca2 + ]i 具有同步的抑制作用 。

Aim To examine the effects of a novel selective phosphodiesterase(PDE) type 3 inhibitors, cilostazol on superoxide free radicals(O · 2)generation and calcium concentration( i)in neutrophils. Methods Cultured HL-60 cells were induced by 1.3% DMSO to differentiate into neutrophils-like cells. Simultaneous measurements of O · 2 production and i were performed with a novel monitor by detecting chemiluminescence and fluorescence of samples at the same time, and the effects of cilostazol on fMLP-stimulated neutrophils were investigated. Results Cilostazol (1～30 μmol·L -1) inhibited fMLP-stimulated O · 2 generation as well as i increase in neutrophils almost at the same time, and especially at the concentrations of 10 μmol·L -1 and 30 μmol·L -1, the differences between the cilostazol groups and the control showed significant statistic meaning (P<0.01). On the other hand, fMLP-induced O · 2 generation was later than i increase in time-phase, and 10 μmol·L -1 cilostazol exhibited synchronous effects on both O · 2 and i. Conclusion Cilostazol effectively regulates the functions of fMLP-stimulated neutrophils by controlling O · 2 generation and levels of i, which indicates that the pharmacology effects of the agent might partially be via a neutrophils-dependent pathway.