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目的探讨地塞米松对哮喘患者单核细胞凋亡状态及其分泌细胞因子的影响作用。方法采用Ficoll-Hypaque密度梯度离心及贴壁培养法对30例支气管哮喘患者和15名健康对照者单核细胞进行分离、纯化后,以空白组、地塞米松组培养24h,经琼脂糖凝胶电泳、流式细胞仪检测细胞凋亡的发生率,同时以酶联免疫吸附试验(ELISA)检测细胞培养上清液中粒细胞—巨噬单核细胞集落刺激因子(GM-CSF)、肿瘤坏死因子(TNF)-α水平。结果哮喘组单核细胞自然凋亡率明显低于对照组(P<0.01),分泌TNF-α、GM-CSF较对照组明显增高(P<0.01)。地塞米松作用后,哮喘组单核细胞凋亡率明显增加(P<0.05);细胞因子分泌水平下降,两组间比较差异无显著性(P>0.05)。结论地塞米松促进哮喘患者单核细胞凋亡并抑制其分泌前炎性细胞因子TNF-α、GM-CSF可能是其治疗支气管哮喘的机制之一。
Objective To investigate the effect of dexamethasone on monocyte apoptosis and secretion of cytokines in asthmatic patients. Methods The mononuclear cells of 30 patients with bronchial asthma and 15 healthy controls were isolated by Ficoll-Hypaque density gradient centrifugation and adherent culture. After purification, the monocytes were cultured in the blank group and the dexamethasone group for 24 h. The cells were separated by agarose gel The incidence of apoptosis was detected by electrophoresis and flow cytometry. The levels of GM-CSF, tumor necrosis factor-α (TNF-α) in cell culture supernatants were detected by enzyme linked immunosorbent assay (ELISA) Factor (TNF) -α levels. Results The natural apoptosis rate of monocytes in asthma group was significantly lower than that in control group (P <0.01). The secretion of TNF-α and GM-CSF in asthma group was significantly higher than that in control group (P <0.01). After dexamethasone treatment, the apoptosis rate of monocytes in asthma group was significantly increased (P <0.05), while the secretion of cytokines decreased. There was no significant difference between the two groups (P> 0.05). Conclusion Dexamethasone can promote monocyte apoptosis and inhibit the secretion of proinflammatory cytokines TNF-α and GM-CSF in asthma patients, which may be one of its mechanisms in the treatment of bronchial asthma.