目的研究急性期蛋白成分纤维蛋白原对中性粒细胞分泌白细胞介素(IL)-1β和IL-8水平的影响,探讨纤维蛋白原在牙周组织炎性反应破坏中的作用。方法采用密度梯度离心分离法分离纯化、体外培养人外周血中性多形核白细胞,通过夹心ELISA法测定加入不同浓度外源性人纤维蛋白原后中性多形核白细胞分泌IL-1β和IL-8的水平。结果加入2g/L纤维蛋白原后中性多形核白细胞于不同时间分泌IL-1β和IL-8的水平分别为(10.41±0.37)~(35.86±0.30)ng/L和(93.90±13.95)~(2045.66±53.03)ng/L;加入10g/L纤维蛋白原后中性多形核白细胞于不同时间分泌IL-1β和IL-8的水平分别为(22.81±0.45)~(57.77±2.08)ng/L和(115.02±10.61)~(3858.69±25.65)ng/L。与阴性和阳性对照组比较,中性多形核白细胞分泌IL-1β(P〈0.001)和IL-8(P≤0.016)的水平显著升高;随加入纤维蛋白原浓度的升高和作用时间的延长,分泌IL-1β和IL-8的水平升高(P〈0.001)。结论纤维蛋白原可促进中性多形核白细胞分泌促炎细胞因子,在增强炎性反应和对牙周组织的破坏中可能发挥重要的病理作用。
Objective To investigate the effect of one of the acute-phase proteins, fibrinogen, on the release of IL-1β and -8 by human peripheral polymorphonuclear leukocytes(PMN) and the possible role of fibrinogen during the destruction of periodontium. Methods Peripheral PMN were isolated by discontinuous density gradient centrifuging technique. The freshly isolated PMN were suspended in Hank's balanced saline solution ( 1 × 10^9/L) supplemented with 0. 5% BSA and 0. 1% glucose. The levels of IL-1β and -8 in the supernatants produced by cultured cells upon the addition of human fibrinogen at different concentrations were measured by ELISA technique. Results Incubated with human fibrinogen at 2 g/L or 10 g/L for different time periods, human peripheral PMN released significantly greater amount of IL-1β [ ( 10. 41 ±0. 37) -(35.86 ±0. 30) ng/L or (22. 81 ± 0. 45 ) -(57. 77 ± 2. 08 ) ng/L] and IL-8 [ (93.90 ± 13.95 )- (2045.66 ± 53.03 ) ng/L or ( 115.02 ± 10. 61 )- (3858.69 ± 25.65 ) ng/L ] than PMN without the stimulation of fibrinogen (IL-1β,P 〈0. 001, and IL-8, P≤0. 016). The higher concentration of fibrinogen or the longer treatment time, the higher levels of IL-1β and -8 were released by PMN (P 〈 0. 001 ). Conclusions Fibrinogen induced the secretion of pro-inflammatory cytokines IL-1β and -8 by PMN and may be involved in magnification of the inflammatory response of periodontium and bone resorption.