雷盛钦+胡水婷+程丽+黄小燕

[摘要]目的 探讨低剂量脂多糖（LPS）刺激对哮喘小鼠T淋巴细胞产生IL-17的影响及其机制。方法 复制哮喘小鼠模型，体外分离培养脾脏来源的T淋巴细胞，分为对照组和LPS组，其中LPS组又分为LPS1组和LPS2组，分别以1、2 ng/ml的LPS体外刺激T细胞72 h。对照组以生理盐水代替LPS。采用ELISA法检测细胞培养上清液中的IL-17水平，观察IL-17的水平浓度变化。结果 以低剂量LPS刺激哮喘小鼠T淋巴细胞后，其培养上清液中IL-17的水平较对照组明显降低，差异有统计学意义（P<0.01）。结论 低剂量LPS可以抑制哮喘T细胞IL-17的分泌，可能与机体的免疫耐受相关。

[关键词]脂多糖；哮喘小鼠；T细胞；白介素-17

[中图分类号] R-332 [文献标识码] A [文章编号] 1674-4721（2017）01（c）-0120-03

[Abstract]Objective To explore the influence of LPS with low dosage on the secretion of IL-17 and the potential mechanism.Methods The asthma mouse model was copied successfully and the T cells from spleen were isolated and cultured in vitro，and the mice were divided into the control group and the LPS group.The mice in LPS group were divided into the LPS group 1 and the LPS group 2，and were stimulated with 1，2 ng/ml LPS respectively for 72 hours.The control group was stimulated with the same volume of saline.The concentrations of IL-17 in the supernatant were detected with ELISA，the change of concentration of IL-17 was observed.Results After stimulation with lower dosages of LPS，the secretion of IL-17 in the supernatant decreased significantly in the LPS groups than that in the control group，with significant difference （P<0.01）.Conclusion Low dosage of LPS can inhibit the secretion of IL-17 in T cells of asthma，and it may be related with immune tolerance.

[Key words]Lipopolysaccharide；Asthma mice；T cell；Interleukin-17

支氣管哮喘是全球常见的慢性疾病之一，是由T细胞、B细胞、嗜酸粒细胞、肥大细胞等多种炎症细胞和细胞因子参与的慢性气道炎症性疾病[1-2]。T细胞及其亚群如Th2细胞分泌的细胞因子如白介素-4（interleukin-4，IL-4）、IL-5、IL-13、IL-17等直接参与了哮喘的炎症过程，进而诱发哮喘的一系列症状[3]。IL-17是一种主要由CD4+T淋巴细胞、嗜酸粒细胞等分泌的前炎性细胞因子，具有强大的募集中性粒细胞及促进多种炎性因子释放的作用，参与了机体多种炎症性疾病的发生、发展[4-5]，在哮喘的慢性气道炎症过程中，也发挥着重要的作用[6]。本实验利用低剂量脂多糖（lipopolysaccharide，LPS）在体外刺激哮喘小鼠T细胞，检测其IL-17的产生情况，探讨低剂量LPS诱导IL-17的产生及哮喘T淋巴细胞免疫耐受的可能机制。

1材料与方法

1.1主要试剂及仪器

RPMI 1640培养基、胎牛血清（Hyclone公司），Al（OH）3（A8222，Sigma公司），鸡卵清蛋白（A5503，Sigma公司），尼龙毛柱（Cat.146-04231，日本Wako公司），小鼠红细胞裂解液（碧云天公司……