摘要
目的探讨布地奈德吸入给药对慢性支气管哮喘模型小鼠多形螺旋线虫预警素释放抑制剂/转录因子(HpARIh/STAT)信号通路的影响。方法30只小鼠随机分成3组,对照组、模型组和布地奈德组,每组10只。模型组小鼠于第1天和第14天用卵白蛋白(OVA)致敏,第24天开始雾化吸入1%OVA并持续刺激至第28天,建立哮喘气道重塑模型;布地奈德组小鼠在吸入OVA 2 h前吸入布地奈德30 min;对照组使用生理盐水代替OVA。第28天最后一次雾化结束后24 h处死小鼠,采用病理图像分析系统测量小鼠的气道形态学参数,分别取各组小鼠外周血和肺组织,用ELISA试剂盒检测外周血上清液肿瘤坏死因子(TNF-α)、白细胞介素13(IL-13)和转化生长因子-β(TGF-β)水平,HE染色观察肺组织病理学变化,Western blotting检测肺组织蛋白HpARIh、IL-13、STAT3和STAT6的表达。结果病理组织学结果显示布地奈德组小鼠支气管壁的炎性细胞浸润明显减少,支气管壁结构改变明显减轻。HE染色结果显示布地奈德干预后有效地减少了炎性因子对肺组织的浸润。ELISA结果显示布地奈德组与模型组相比,外周血上清中TNF-α、TGF-β和IL-13等炎症因子水平明显降低(P<0.05),但仍高于对照组。Western blotting结果表明布地奈德通过增加HpARIh蛋白表达来激活STAT3,同时抑制STAT6,从而抑制哮喘诱发因子IL-13的表达水平。结论早期雾化吸入布地奈德可以有效抑制TNF-α、TGF-β和IL-13等炎症因子的表达,同时通过激活HpARIh/STAT信号通路抑制IL-13的表达,明显减轻哮喘小鼠的气道重塑。
Objective To investigate the effect of budesonide on HpARIh/STAT signaling pathway in chronic bronchial asthma model mice.Metheds Thirty mice were randomly divided into three groups:normal control group,model group and budesonide group,with 10 rats in each group.Mice in model group were sensitized with ovalbumin(OVA)on day 1 and day 14,and aerosolized 1%OVA on day 24 and continued drug stimulation until day 28 to establish an asthmatic airway remodeling model;mice in budesonide group were inhaled budesonide 30 min before 2 h inhalation of OVA;mice in normal control group used physiological saline instead of OVA.On the 28th day,the mice were sacrificed 24 h after the last atomization.The pathological image analysis system was used to measure the airway morphological parameters of mice.Then the peripheral blood and lung tissues of mice in each group were taken,and the peripheral blood was detected by ELISA kit.The expression levels of tumor necrosis factor(TNF-α),interleukin-13(IL-13)and transforming growth factor-β(TGF-β)were detected in the supernatant.HE staining was used to observe the pathological changes of lung tissue.Western blotting was used to detect protein expression of HpARIh,IL-13,STAT3 and STAT6 in lung tissue.Results Histopathological results showed that the inflammatory cell infiltration of the bronchial wall and the structural changes of the bronchial wall of mice in budesonide group were reduced significantly(P<0.05).HE staining results showed that the infiltration of inflammatory factors in lung tissue of mice in budesonide group were reduced(P<0.05).The results of ELISA showed that the levels of TNF-α,TGF-βand IL-13 in peripheral blood supernatant of mice in budesonide group were significantly lower than in model group(P<0.05),but still higher than control group.Results of Western blotting indicated that budesonide inhibited the expression of the asthma-inducible factor IL-13 by increasing the expression of HpARIh and inhibiting STAT6.Conclusion Early aerosolized inhalation budesonide can effectively inhibit the expression of inflammatory factors such as TNF-α,TGF-βand IL-13,and inhibit the expression of IL-13 by activating HpARIh/STAT signaling pathway,which can significantly reduce asthma.
作者
武国宇
WU Guo-yu(Department of Pharmacy,The First Affiliated Hospital of Dalian Medical University,Dalian 116000,China)
出处
《现代药物与临床》
CAS
2020年第12期2307-2311,共5页
Drugs & Clinic
基金
辽宁省教育厅计划项目(201602223)
辽宁省科技厅攻关项目(2019-ZD-0652)。