蛇床子素预处理对卵清蛋白诱导的小鼠哮喘气道炎症及黏液高分泌的调控作用

Effect of osthole on asthmatic airway inflammation and mucus hypersecretion induced by ovalbumin in mice

目的探讨蛇床子素(Ost)预处理对卵清蛋白(OVA)诱导的哮喘小鼠气道炎症、黏液高分泌的防治作用及其机制。方法将40只Balb/c小鼠随机分为4组:生理盐水(PS)对照组、哮喘模型组、Ost处理组1(10 mg/kg)、Ost处理组2(40 mg/kg)。利用OVA构建哮喘小鼠模型。末次激发48 h后,处死各组小鼠,取肺组织及肺泡灌洗液,利用HE,AB-PAS染色法观察肺组织炎性病变及黏液分泌状况,利用ELISA法检测支气管肺泡灌洗液(BALF)中Th1/2细胞因子表达水平。Real-time-PCR检测各组小鼠肺组织中mCLCA3以及MUC5AC的表达水平。结果哮喘组小鼠肺组织炎症及气道内黏液分泌水平较对照组明显增高,Ost可有效减低OVA所致的气道炎症以及气道黏液分泌;Ost组小鼠支气管肺泡灌洗液中细胞总数及嗜酸性粒细胞数明显降低;Ost对白介素-4(IL-4)、Eotaxin具有明显的抑制作用,同时对干扰素-(IFN-γ)具有提升作用。此外,Ost抑制肺组织中mCLCA3及MUC5AC mRNA的表达。结论 Ost预处理对OVA诱导的小鼠哮喘气道具有保护作用,其机制可能是通过下调mCLCA3 mRNA进而抑制MUC5AC表达,减弱哮喘气道黏液分泌的作用。

Objective To explore the effect of pretreatment osthole （Ost） on ovalbumin-induced asthmatic airway inflammation,mucus hypersecretion in mouse and to study the mechanism.Methods A total of 40 Balb/c mice were divided into 4 groups randomly：physiological saline （PS） control group,asthma model group,Ost group 1（10 mg/kg）,Ost group 2 （40 mg/kg）.Asthma model was set up by ovalbumin （OVA）.All mice were sacrificed 48 h after the last challenge.Lung tissue and bronchial alveolar lavage fluid（BALF） were collected,and HE and AB-PAS stain were used to analysis airway inflammation and mucus secretion.ELISA analysis was used to detect the level of several cytokines in BALF.The mCLCA3 and MUC5AC mRNA level were determined by Real-time PCR.Results Ost could suppress the airway inflammation and mucus secretion effectively.And the total cell number and Eosinophils number of BALF was suppressed by Ost; Ost inhibit the IL-4 and Eotaxin,but elevated IFN-γ.Furthermore,Ost down-regulate the mCLCA3 and MUC5AC mRNA level.Conclusion Ost pretreatment on OVA-induced airway inflammation in mice has a protective effect,the mechanism may be through down mCLCA3 mRNA and inhibit MUC5AC protein levels,reduce reducing airway mucus secretion.