PM2.5对哮喘小鼠气道重塑的作用机制研究
Mechanism of PM2.5 on Airway Remodeling in Asthmatic Mice
李章紊 1高洁 2刘健玲 1王爱利 2李中鹏 2吴健1
作者信息
- 1. 510006 广州,华南理工大学医学院;510080 广州,南方医科大学附属广东省人民医院广东省医学科学院老年呼吸二科
- 2. 510080 广州,南方医科大学附属广东省人民医院广东省医学科学院老年呼吸二科
- 折叠
摘要
目的 探讨PM2.5对卵清蛋白(OVA)诱导的哮喘小鼠气道重塑的影响,并分析其潜在机制.方法 将36 只SPF级雌性BALB/c小鼠随机分为对照组、哮喘组、哮喘+ PM2.5 组,每组 12 只.哮喘组、哮喘+ PM2.5 组于第 1、7、14 天腹腔注射OVA致敏,第21～27 天连续进行1%OVA雾化激发,每次30 min,共7 次.其中哮喘+ PM2.5组每次于OVA激发前30 min给予100 μg PM2.5 混悬液滴鼻干预.对照组用生理盐水处理.3 组均在末次激发24h后进行气道阻力检测,处死动物后收集小鼠肺泡灌洗液(BALF)及肺组织标本.通过HE、PAS和Masson染色分别评估小鼠肺部炎症、杯状细胞增生和胶原纤维沉积情况,免疫组织化学法检测肺组织α-平滑肌肌动蛋白(α-SMA)的表达;通过ELISA法检测小鼠BALF中白细胞介素-4(IL-4)和IL-5 水平;通过Western blot法检测小鼠肺组织p38 丝裂原活化蛋白激酶(p38 MAPK)/核因子-κB(NF-κB)信号通路相关蛋白表达.结果 与哮喘组比较,哮喘+ PM2.5 组气道阻力及炎症评分升高,杯状细胞增多,胶原纤维增生明显,BALF中IL-4、IL-5 水平也显著增高(P＜0.05).与哮喘组比较,哮喘+ PM2.5组肺组织中转化生长因子-β1(1.25±0.17 vs 0.87±0.25)、p-p38 MAPK/p38 MAPK(1.24±0.24 vs 0.74±0.19)、p-NF-κB p65/NF-κB p65(1.02±0.14 vs 0.70±0.10)蛋白表达水平升高(P＜0.05).结论 PM2.5 可通过激活p38 MAPK/NF-κB信号通路加重哮喘小鼠的气道重塑.
Abstract
Objective To investigate the effect of PM2.5 on airway remodeling induced by ovalbumin(OVA)in asth-matic mice,and to analyze its underlying mechanism.Methods In total,36 SPF female BALB/c mice were randomly divid-ed into control group,asthma group and asthma +PM2.5 group,with 12 mice in each group.Mice in the asthma group and the asthma +PM2.5 group were sensitized by intrabitoneal injection of OVA on days 1,7 and 14,and were stimulated by 1%OVA atomization on days 21-27(30 min each time for a total of 7 times).Mice in asthma +PM2.5 group were given 100 μg PM2.5 suspension nasal drops at 30 min before each OVA stimulation.The control group was treated with normal saline.Airway re-sistance was detected at 24 h after the last stimulation of mice in the three groups.Bronchoalveolar lavage fluid(BALF)and lung tissue samples were collected after the animals were sacrificed.Hematoxylin-eosin(HE)staining,PAS staining and Masson staining were used to evaluate the pulmonary inflammation,trophy cell hyperplasia and collagen fiber deposition.The expression of α-smooth muscle actin(α-SMA)in lung tissue was detected by immunohistochemistry,and the levels of inter-leukin-4(IL-4)and interleukin-5(IL-5)in BALF were determined by ELISA.The expression level of p38 mitogen-activated protein kinase(p38 MAPK)/nuclear factor-κB(NF-κB)signaling pathway related proteins in mouse lung tissues was detec-ted by Western blot.Results Compared with the asthma group,the airway resistance and inflammation scores of asthma + PM2.5 group were increased,Goblet cells were increased,the collagen fiber hyperplasia was obvious,and the levels of IL-4 and IL-5 in BALF were also significantly increased(P＜0.05).Compared with the asthma group,the protein expression lev-els of transforming growth factor-β(1.25±0.17 vs.0.87±0.25),p-p38 MAPK/p38 MAPK(1.24±0.24 vs.0.74±0.19),p-NF-κB p65/NF-κB p65(1.02±0.14 vs.0.70±0.10)in lung tissue of asthma + PM2.5 group were increased(P＜0.05).Conclusion PM2.5 can aggravate airway remodeling in asthmatic mice by activating the p38 MAPK/NF-κB sig-naling pathway.
关键词
PM2.5/哮喘/气道重塑/p38丝裂原活化蛋白激酶类/核因子-κB/杯状细胞/白细胞介素-4/小鼠,BALB/cKey words
PM2.5/Asthma/Airway remodeling/p38 mitogen-activated protein kinases/NF-κB/Goblet cells/In-terleukin-4/Mice,BALB/c引用本文复制引用
基金项目
国家自然科学基金资助项目(面上项目)(81970012)
出版年
2023
国家科技期刊平台
NSTL
万方数据