探讨雷公藤甲素对BALB/c哮喘小鼠IL-23/Th17(IL-17)炎症轴的影响

To explore effects of triptolide on IL-23 / Th17( IL-17) inflammatory axis in asthmatic BALB / c mice

目的:观察雷公藤甲素对哮喘小鼠IL-23、Th17细胞及其细胞因子IL-17表达的情况,探讨其对Th17细胞介导的哮喘气道炎症的影响及其作用机制,为雷公藤甲素治疗哮喘提供作用靶点。方法:SPF级BALB/c小鼠32只,随机分为正常组(NC组)、哮喘组(A组)、哮喘雷公藤甲素干预组(TA组)和哮喘地塞米松干预组(DA组),每组8只。哮喘组以卵白蛋白和氢氧化铝致敏;卵白蛋白滴鼻吸入激发。雷公藤甲素干预组及地塞米松干预组致敏及激发方法均同哮喘组,这两组在每次激发前30 min分别予腹腔注射雷公藤甲素及地塞米松,正常对照组致敏与激发均以生理盐水代替。细胞计数器计算肺泡灌洗液(BALF)中白细胞总数及嗜酸性粒细胞的数目;酶联免疫吸附试验(ELISA)检测小鼠BALF IL-23,IL-17的含量;HE染色观察小鼠肺组织病理变化;免疫组化法检测肺组织IL-17蛋白表达水平;q RT-PCR检测右肺组织中IL-17m RNA的表达;流式细胞仪检测外周血Th17细胞占CD4+T淋巴细胞百分率。结果:哮喘组肺泡灌洗液(BALF)中白细胞(WBC)、嗜酸性粒细胞(Eos)、IL-23、IL-17的含量、肺组织中IL-17蛋白及IL-17 m RNA表达、外周血Th17细胞占CD4+T细胞的比率均高于对照组(P<0.05),雷公藤甲素干预组和地塞米松干预组中上述各指标低于哮喘组(P<0.05);雷公藤甲素干预组与地塞米松干预组上述各指标间差异无显著性(P>0.05)。结论:雷公藤甲素可抑制哮喘气道炎症,其作用机制可能通过抑制IL-23/Th17(IL-17)炎症轴实现。

Objective：To observe the effect of triptolide on asthmatic mice IL-23, Th17 cells and their cytokine IL-17 expression,and to explore its effect on Th17 cell-mediated airway inflammation,and its mechanism of action,which provides targets for triptolide in treatment of asthma.Methods： 32 SPF level BALB/c mice were randomly divided into normal control group （ NC ） , asthmatic group （ A ） , triptolide group （ TA group ） and dexamethasone group （ DA group ） , n=8.Asthmatic group with ovalbumin sensitization and aluminum hydroxide;ovalbumin intranasal inhalation challernge.Mice of triptolide group and dexamethasone group were sensitized and challenged as asthmatic group, and the two groups were respectively given triptolide and dexamethasone by intraperitoneal injection 30 minutes before challenged.Mice of control group was sensitized and challenged by saline.The total number of white blood cells and the number of eosinophils of BALF were calculated by cell counter.IL-23 and IL-17 levels in BALF were measured by ELISA.Lung tissue were stained with hematoxyin and eosin（HE）.IL-17 protein expression levels were detected by immu-nohistochemistry in lung tissue,and the mRNA expression levels of right lung tissue were detected by qRT-PCR.Th17 percentage of CD4＋T lymphocytes were analyzed by flow cytometry.Results：Numbers of white blood cells（ WBC） and eosinophils（ Eos） of BALF, IL-23 and IL-17 levels of BALF,IL-17 protein and IL-17 mRNA expression in lung tissue,and Th17 cell frequencies in peripheral blood were all significantly increased in the asthmatic group compared to the control group（P〈0.05）,but reduced significantly in triptolide group and dexamethasone group compared to asthmatic “there was no significant difference in the above mentioned indicators between in triptolide group and dexamethasone group （ P〈0.05 ） .Conclusion： Triptolide can inhibit airway inflammation, which”nbsp;mechanism is possible by inhibiting IL-23/Th17（IL-17） inflammatory axis.