摘要
目的:探索预防性IL-27滴鼻给药治疗哮喘小鼠气道炎症的最佳干预方式,并直接从组织蛋白水平进行相关研究。方法:将96只雌性C57/6J小鼠随机分成对照组、哮喘组、IL-27滴鼻预防组Ⅰ和IL-27滴鼻预防组Ⅱ。在卵白蛋白(OVA)腹腔注射致敏与滴鼻激发诱导的小鼠哮喘模型基础上,加用IL-27干预,一种为"致敏前IL-27小剂量、多次预防模型",一种为"致敏后激发前IL-27小剂量、多次预防模型"。取半数小鼠激发后处死,做气道炎症水平评估;取半数小鼠,激发前一天处死进行肺组织信号转导与转录激活子1(STAT1)和STAT1磷酸化水平的Western blot检测。结果:致敏前IL-27小剂量、多次预防给药,可以明显降低哮喘小鼠BAL中IL-5和IL-13的水平(P<0.05),抑制支气管及血管周围的炎症细胞浸润,炎症评分明显低于哮喘组(P<0.05),而激发前致敏后给药则无明显的统计学意义(P>0.05)。哮喘小鼠体内STAT1的磷酸化在激发前就已明显受损,致敏前的IL-27给药能扭转这种损伤,而激发前致敏后给药则不能。结论:致敏前IL-27小剂量、多次预防给药通过逆转哮喘小鼠体内STAT1磷酸化的受损能明显改善哮喘小鼠的气道炎症,而致敏后激发前的小鼠则由于在致敏阶段STAT1磷酸化就已受损而对IL-27的作用产生耐受,这种耐受可能是由于致敏后小鼠气道的CD4+T细胞就已大部分转化为Th2细胞。
Objective： Animal models were set up to explore the best preventative intra-nasal administration of interleukin 27（ IL-27） to diminish allergic airway inflammation of asthma and the related molecular mechanisms. Methods： Ninety-six female C57 /6J mice were randomly divided into four groups,a group of the control group,a group of asthma group,and two groups of the prevention group. Based on being sensitizing and challenging with Ovalbumin（ OVA） in the asthma model,two kinds of IL-27 administration asthma animal models were set up,one of which was low-dose-multiple preventive administration before OVA sensitization,one was lowdose-multiple preventive administration after OVA sensitization but before OVA challenge. Sacrificed the mice after challenging and analyzed the IL-5 and IL-13 levels in supernatant of Broncho alveolar lavage fluid（ BAL） using ELISA; and HE stain and inflammation score were done for the lungs. Sacrificed the mice before challenging and used the lungs to analyze the level of total signal transducer and activator of transcription-1（ STAT1） protein and phos-STAT1 based on the method of Western blot. Results： In low-dose-multiple administration before sensitization preventions group,IL-27 inhibits the secretion of IL-5 and IL-13（ P〈0. 05） and inflammation around bronchial and vascular obviously,and the inflammation score was lower than asthma group（ P〈0. 05）,while another group has no significant effects（ P〉0. 05）. The phosphorylation of STAT1 was impaired in mice after OVA sensitization,and preventative administration of IL-27 before sensitization could reverse the impairment of STAT1,whereas another group had no obviously changes. Conclusion： Preventative administration of IL-27 before sensitization can attenuate the airway inflammation in the mouse asthma model via reversing the phosphorylation of STAT1,while the mice which has been sensitized resisting the inhibition of IL-27 due to the impairment of the phosphorylation of STAT1 and already committed Th2-CD4＋T cells existed in sensitization-mice airway might be the reason for such IL-27 resistance.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2016年第7期1022-1027,共6页
Chinese Journal of Immunology
基金
国家自然科学基金(No.81270078和No.81470211)项目资助
