实验性变应性鼻炎小鼠脾细胞培养上清细胞因子测定及其变化特征

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目的:探讨变应性鼻炎小鼠模型脾细胞在不同刺激物刺激下培养上清中细胞因子在不同时间段的水平,比较2种刺激物对细胞因子的影响效果,以及在不同时间段细胞因子的分泌情况。方法:用卵清蛋白(OVA)致敏小鼠,并用OVA盐溶液滴鼻激发制成变应性鼻炎模型,对照组用生理盐水替代,在末次激发后取脾单核细胞培养,并分别加入OVA和刀豆蛋白(ConA)刺激脾单核细胞增殖、分泌细胞因子,分别于培养后24、48、72h取培养上清液进行细胞因子IL-4、IL-5、IFN-γ浓度测定。结果:实验组脾单核细胞培养上清中加ConA刺激产生IL-4、IL-5和IFN-γ浓度均明显高于对照组(均P<0.05),但IFN-γ/IL-4、IL-5明显降低;加OVA刺激产生的IL-4、IL-5比对照组明显增高但升高幅度比加入ConA的低,而IFN-γ浓度则明显降低(P<0.05)。脾细胞培养上清中IL-4含量在72h内随时间持续增高,IL-5及IFN-γ在48h内增长显著,48~72h内增长趋缓。结论:小鼠实验性变应性鼻炎体内T细胞被广泛激活,受抗原OVA和非特异性刺激物ConA刺激Th2细胞释放的细胞因子明显增多,而Th1细胞及其细胞因子则受抑制;ConA刺激产生的细胞因子差异更显著。在2种刺激物刺激下72h内各细胞因子是持续增多的。 Objective: To investigate the levels of cytokines in supernatants of splenocytes of mice with allergic rhinitis induced by different stimuli at different time periods, and to compare the effect of two stimuli on cytokines and the effects of cytokines The secretion of. Methods: Mice were sensitized with ovalbumin (OVA) and intranasally challenged with OVA salt solution to make the model of allergic rhinitis. The control group was replaced by normal saline. After the last challenge, spleen mononuclear cells were cultured and added OVA and concanavalin (ConA) stimulated spleen mononuclear cells to proliferate and secrete cytokines. The culture supernatants were harvested at 24,48,72 h after culture to determine the concentrations of cytokines IL-4, IL-5 and IFN-γ. Results: The concentrations of IL-4, IL-5 and IFN-γ in the culture supernatant of spleen mononuclear cells treated with ConA in the experimental group were significantly higher than those in the control group (all P <0.05) IL-5, IL-5 and IL-5 in OVA-stimulated group were significantly higher than those in control group, but lower than those in ConA group, while IFN-γ concentration was significantly decreased (P <0.05). The content of IL-4 in the spleen cell culture supernatant continued to increase with time within 72h, the IL-5 and IFN-γincreased significantly in 48h, but slowed in 48 ~ 72h. CONCLUSIONS: T cells are widely activated in mice with experimental allergic rhinitis. The release of cytokines by Th2 cells stimulated by antigen OVA and non-specific stimulator ConA is significantly increased, whereas Th1 cells and cytokines are inhibited. ConA stimulation produces The differences in cytokines were even more pronounced. In the two stimuli stimulated within 72h of each cytokine is continuously increased.
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