抗类风湿病药金诺芬对中性粒细胞自发性凋亡的影响

Effect of auranofin, an antirheumatoid drug, on neutrophil apoptosis

目的 研究抗类风湿病药金诺芬(AF)对中性粒细胞自发性凋亡的影响。方法 运用流式细胞技术和MTS细胞活性测定技术对不同浓度的金诺芬影响下的中性粒细胞活性及其凋亡进行观察,并用低分子质量DNA“梯形条带”(DNA ladder)检测技术对其凋亡进行鉴定。结果 低浓度金诺芬(1μmol/L)可通过延迟中性粒细胞的自发性凋亡而增加其生命期限;但当金诺芬浓度大于5μmol/L时,中性粒细胞的坏死则明显增加,因而缩短了其生命期限。结论 金诺芬临床治疗量时的血清浓度一般在0.7μmol/L左右,在此浓度下,中性粒细胞通过延迟自发性凋亡来延长其功能性生命期限,所以金诺芬对类风湿关节炎患者的抗炎、抗风湿作用不可能通过直接清除炎症局部的中性粒细胞来完成;金诺芬的抗炎、抗风湿作用可能存在更为复杂的作用机制。

Objective To investigate the effects of auranofin (AF) on apoptosis of neutrophils. Methods Neutrophils were incubated with various concentrations of AF for different periods. Cell viability was determined by the MTS assay and apoptosis was evaluated by flow cytometric analysis of propidium iodide (PI)-staining of the nuclei and annexin-V staining of phosphatidylserine in the cell membrane. Results At a low concentration (1 μmol/L), AF significantly prolonged neutrophil survival by delaying spontaneous apoptosis. In contrast, a higher AF concentration (5μmol/L) shortened neutrophil survival by the induction of cell necrosis. Conclusion The serum concentration of AF in patients with rheumatoid arthritis is about 0.7μmol/L, in which neutrophils are prolonged their functional life-span through delaying the spontaneous apoptosis. Thus, it seems to be unlikely that AF exerts the anti-inflammatory effect in vivo by directly suppressing neutrophil functions. The therapeutic benefit of AF in rheumatoid arthritis may be mediated in a complex manner.