摘要
目的:探讨清肺化痰汤对哮喘小鼠Clara细胞向杯状细胞化生的影响。方法:将60只C57BL/6J小鼠随机分为正常对照组、模型对照组、清肺化痰汤10.5、21.0、42.0 g/kg组及盐酸氨溴索49 mg/kg组,每组10只。第1、8 d除正常对照组外小鼠腹腔注射0.5 mL OVA混悬液(含100μg OVA,200μg氢氧化铝),第15 d~21 d雾化,雾化结束1 h后给药,连续给药7 d,末次给药24 h后,部分动物以小动物肺功能仪检测小鼠肺功能增强的呼气间歇;取血,检测血常规;部分小鼠立即处死,取肺组织,HE染色观察小鼠肺组织炎症程度及病理变化,Masson染色检测气道重塑情况,AB-PAS染色检测气道黏液分布,Real-time RT-qPCR检测小鼠肺组织Clara细胞分泌蛋白(Clara cell secretory protein,Ccsp)及黏蛋白5AC(Mucin 5AC,Muc5ac)mRNA的表达,Western blot检测小鼠肺组织中CCSP、MUC5AC蛋白的表达。结果:与正常对照组比较,模型对照组小鼠增强的呼气间歇显著升高(P<0.01);血中嗜酸性粒细胞含量明显升高(P<0.05);肺组织形态改变、肺泡壁变厚;胶原沉积含量显著增加;远端气道黏液分泌升高;肺组织中Muc5ac mRNA表达显著上调,Ccsp mRNA表达显著下调;肺组织中MUC5AC蛋白表达显著上调,CCSP蛋白表达显著下调(P<0.01)。与模型对照组比较,各给药组小鼠增强的呼气间歇显著降低(P<0.01);清肺化痰汤21.0、42.0 g/kg血中嗜酸性粒细胞含量明显降低(P<0.05);肺组织形态明显改善、肺泡壁变薄;清肺化痰汤10.5、21.0、42.0 g/kg胶原沉积含量显著下降(P<0.01);远端气道黏液含量显著降低(P<0.01);Ccsp mRNA表达显著上调(P<0.05或P<0.01),清肺化痰汤21.0、42.0 g/kg肺组织中Muc5ac mRNA表达显著下调(P<0.01),清肺化痰汤10.5、42.0 g/kg肺组织中MUC5AC蛋白表达显著上调(P<0.01),清肺化痰汤42.0 g/kg CCSP蛋白表达明显下调(P<0.05)。结论:清肺化痰汤可能通过抑制Clara细胞向杯状细胞化生,减轻黏液高分泌从而治疗哮喘。
Objective:To explore the effect of Qingfei Huatan Decoction on the Clara cells from differentiating into goblet cells in asthma mice.Methods:Sixty C57 BL/6 J mice were randomly divided into a control group,a model group,low-,medium-,and high-dose Qingfei Huatan Decoction groups(10.5,21.0,and 42.0 g/kg),and an ambroxol hydrochloride group(49 mg/kg),with 10 mice in each group.On the 1 st and 8 th days,except those in the control group,mice were injected intraperitoneally with 0.5 mL of OVA suspension(containing 100μg of OVA and 200μg of aluminum hydroxide),followed by nebulization from the 15 th to 21 th days.One hour after nebulization,corresponding drugs were provided for 7 days.Twenty-four hours after the last administration,a small-animal pulmonary function detector was used to detect the enhanced pause(Penh)of improved lung function on some mice,and blood samples were collected for blood routine examination.Some mice were immediately sacrificed,and the lung tissues were taken to observe the degree of inflammation and pathological changes by HE staining.Masson staining was used to detect airway remodeling,and AB-PAS staining was used to detect airway mucus distribution.Real-time RT-qPCR and Western blot were used to detect the mRNA and protein expression of Clara cell secretory protein(CCSP)and mucin 5 AC(MUC5 AC),respectively.Results:Compared with the control group,the model group showed increased Penh(P<0.01),increased eosinophil content in the blood(P<0.05),altered lung tissue morphology,thicker alveolar wall,increased collagen deposition content,improved distal airway mucus secretion,elevated mRNA expression of MUC5 AC,decreased mRNA expression of CCSP,up-regulated protein expression of MUC5 AC,and down-regulated protein expression of CCSP in lung tissues(P<0.01).Compared with the model group,the groups with drug intervention showed reduced Penh of mice(P<0.01).To be specific,the medium-and high-dose Qingfei Huatan Decoction groups showed reduced eosinophil content in the blood(P<0.05),improved lung tissue morphology,and thinned alveolar wall.All the Qingfei Huatan Decoction groups showed down regulated collagen deposition content(P<0.01),decreased distal airway mucus content(P<0.01),and up regulated mRNA expression of CCSP(P<0.05 or P<0.01).In the medium-and high-dose Qingfei Huatan Decoction groups,the mRNA expression of MUC5 AC was down regulated(P<0.01).In the low-and high-dose Qingfei Huatan Decoction groups,the expression of MUC5 AC protein in lung tissues was up-regulated(P<0.01),while the expression of CCSP protein was down-regulated in the high-dose Qingfei Huatan Decoction group(P<0.05).Conclusion:Qingfei Huatan Decoction may treat asthma by inhibiting Clara cells from differentiating into goblet cells and reducing mucus hypersecretion.
作者
屈飞
黄俊浩
席超
张明月
刘婕
王心怡
曾杰
崔艳茹
Qu Fei;Huang Junhao;Xi Chao;Zhang Mingyue;Liu Jie;Wang Xinyi;Zeng Jie;Cui Yanru(Dept of Pharmacology,School of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang 330004;Dept of Physiology,School of Traditional Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第1期145-150,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家自然基金项目(编号:81760744)
江西省教育厅重点项目(编号:GJJ190633)
江西中医药大学创新基金项目(编号:JZYC21S41)
作者简介
通信作者:崔艳茹,副教授,硕士生导师,硕士,研究方向:中药药理学研究,E-mail:cuiyanru2@163.com;屈飞,副教授,硕士生导师,博士,研究方向:中药药理学研究,E-mail:quf0917@163.com。
