金水六君煎对慢性阻塞性肺疾病小鼠气道黏液潴留的影响及机制研究  

Effect and mechanism of Jinshui Liujun decoction on airway mucus retention in COPD mice

作　　者：张真 杨亿然 王慧 李剑庆 李爱平 柏正平[2] 刘雨[3] ZHANG Zhen;YANG Yiran;WANG Hui;LI Jianqing;LI Aiping;BAI Zhengping;LIU Yu(Hunan University of Chinese Medicine,Changsha 410208,Hunan,China;Hunan Academy of Chinese Medicine,Changsha 410006,Hunan,China;The Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine,Changsha 410006,Hunan,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省中医药研究院,湖南长沙410006 [3]湖南省中医药研究院附属医院,湖南长沙410006

出　　处：《现代中西医结合杂志》2023年第13期1753-1761,共9页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金项目(82004306)。

摘　　要：目的 探讨金水六君煎对慢性阻塞性肺疾病(COPD)小鼠气道黏液潴留的影响及其可能机制。方法 将72只C57BL/6小鼠随机分为空白组、COPD组、金水六君煎低剂量组、金水六君煎中剂量组、金水六君煎高剂量组和罗氟司特组,每组12只。除空白组外,其余组采用鼻腔滴注脂多糖加烟熏的方法建立COPD模型。成模后,金水六君煎低、中、高剂量组分别给予3.705 g/(kg·d)、7.41 g/(kg·d)、14.82 g/(kg·d)金水六君煎液灌胃,罗氟司特组给予65μg/(kg·d)罗氟司特灌胃,空白组和COPD组给予等量生理盐水灌胃,均1次/d,连续14 d。灌胃结束后,采用小动物肺功能仪测定各组小鼠每分钟通气量(MV)、呼气峰值流速(PEF)和吸气峰值流速(PIF),HE染色半定量评估肺组织炎症,PAS染色观察气道黏液分泌和杯状细胞增生情况,比色法检测肺泡灌洗液中唾液酸、尿素含量,并计算两者比值,ELISA法检测肺组织中环腺苷酸(cAMP)和V-ATP酶含量及肺泡灌洗液中黏蛋白5ac(Muc5ac)含量,qRT-PCR和Western blot法分别检测肺组织中囊性纤维化跨膜转导调节因子(CFTR)、Foxi1 mRNA和蛋白表达情况。结果 与空白组比较,COPD组的MV、PEF、PIF、尿素含量、cAMP和V-ATP酶含量及CFTR、Foxi1 mRNA和蛋白相对表达量均明显降低(P均<0.05),肺组织病理评分、气道阳性着色面积占比和唾液酸、Muc5ac含量及唾液酸/尿素均明显升高(P均<0.05)。与COPD组比较,金水六君煎各组(除金水六君煎低剂量组cAMP外)和罗氟司特组的MV、PEF、PIF、尿素含量、cAMP和V-ATP酶含量及CFTR、Foxi1 mRNA和蛋白相对表达量均明显升高(P均<0.05),肺组织病理评分、气道阳性着色面积占比和唾液酸、Muc5ac含量及唾液酸/尿素均明显降低(P均<0.05)。结论 金水六君煎可改善COPD小鼠气道黏液潴留,其机制可能与调控CFTR和Foxi1表达有关。Objective It is to explore the effect and possible mechanism of Jinshui Liujun decoction on airway mucus retention in mice with chronic obstructive pulmonary disease(COPD).Methods Seventy-two C57BL/6 mice were randomly divided into blank group,COPD group,low dose group of Jinshui Liujun decoction,medium dose group of Jinshui Liujun decoction,high dose group of Jinshui Liujun decoction,and roflukast group,with 12 mice in each group.All the mice in each group except for the blank group were given lipopolysaccharide by nasal drip plus smoking to established COPD models.After modeling,the low,medium,and high dose groups of Jinshui Liujun decoction were respectively given 3.705 g/(kg·d),7.41 g/(kg·d),and 14.82 g/(kg·d)Jinshui Liujun decoction solution by gavage,while the roflukast group was given 65μg/(kg·d)roflukast by gavage,the blank group and COPD group were given an equal amount of physiological saline by gavage,all once daily,continuously treated for 14 days.After treatment,the minute ventilation volume(MV),peak expiratory flow(PEF)and peak inspiratory flow(PIF)of the mice in each group were detected by a small animal lung function instrument,the lung tissue inflammation was semi quantitatively evaluated by HE staining,the airway mucus secretion and goblet cell proliferation were observed by PAS staining,and the contents of sialic acid and urea in alveolar lavage fluid were measured by colorimetry,and their ratio was calculated,the contents of cyclic adenosine monophosphate(cAMP)and V-ATPase in lung tissue,as well as the content of mucin 5ac(Muc5ac)in alveolar lavage fluid were measured by ELISA,the mRNA and protein expressions of cystic fibrosis transmembrane transduction regulatory factor(CFTR)and forkhead box protein i1(Foxi1)in lung tissue were respectively detected by qRT PCR and Western blot.Results Compared with the blank group,the MV,PEF,PIF,contents of urea,cAMP,V-ATPase,as well as the relative expressions of CFTR,Foxi1 mRNA and protein in the COPD group were significantly reduced(all P<0.05),while

关 键 词：金水六君煎 慢性阻塞性肺疾病 气道黏液潴留 囊性纤维化跨膜转导调节因子 Foxi1 罗氟司特 

分 类 号：R-332[医药卫生]