金水六君煎通过纠正氧化应激介导的CFTR获得性缺陷改善COPD小鼠气道病变  

Jinshu Liujun Decoction Ameliorates Airway Lesions in COPD Mice by Correcting Oxidative StressMediated CFTR Acquired Defects

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(Graduate School of Hunan University of Chinese Medicine,Changsha 410208,China;Hunan Academy of Chinese Medicine,Changsha 410006,China;Hunan Academy of Chinese Medicine Affiliated Hospital,Changsha 410006,China)

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

出　　处：《世界科学技术-中医药现代化》2024年第7期1793-1801,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会青年科学基金项目(82004306):基于肺离子细胞表达的CFTR探讨慢阻肺气道黏液高分泌机制及金水六君煎干预作用,负责人:刘雨。

摘　　要：目的观察金水六君煎对慢性阻塞性肺疾病(COPD)小鼠气道病变的影响,并探讨其可能的机制。方法48只C57BL/6小鼠随机分为空白组、模型组、金水六君煎组和N-乙酰半胱氨酸(NAC)组,每组12只。采用鼻腔滴注脂多糖(LPS)加烟熏的方法建立COPD小鼠模型,成模后予相应药物灌胃14天。观察小鼠的一般情况,小动物肺功能仪测定小鼠每分钟通气量(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、肺泡间隔厚度和气道管壁厚度,过碘酸-雪夫(PAS)染色观察气道黏液分泌和杯状细胞增生情况。试剂盒检测肺泡灌洗液(BALF)中髓过氧化氢酶(MPO)、唾液酸(SA)和尿素(Urea)含量,并计算SA和Urea比值,ELISA法检测BALF中黏蛋白5AC(MUC5AC)含量。试剂盒检测肺组织中活性氧(ROS)、还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)和谷胱甘肽还原酶(GR)水平,并计算GSH和GSSG比值。定量逆转录PCR(RT-qPCR)检测肺组织中囊性纤维化跨膜转导调节因子(CFTR)mRNA表达水平,Western blot检测肺组织中CFTR蛋白表达水平。结果与对照组比较,模型组小鼠生长状况不佳,在造模期间各个时间点的体重均降低(P<0.01),MV、PEF和PIF指标下降(P<0.01),肺组织病理评分、肺泡间隔厚度、气道管壁厚度、气道黏液和杯状细胞均增加(P<0.01),BALF中的SA、SA/Urea、MUC5AC和MPO水平升高(P<0.01),以及Urea水平降低(P<0.01),肺组织中的ROS和GSSG水平升高(P<0.01),以及GSH、GSH/GSSG和GR水平降低(P<0.01)。肺组织中的CFTR mRNA和蛋白表达水平减少(P<0.01)。与模型组比较,COPD小鼠生长状况好转,在造模期间各个时间点的体质量均增加(P<0.05,P<0.01),MV、PEF和PIF指标得到明显改善(P<0.01),肺组织病理评分、肺泡间隔厚度、气道管壁厚度、气道黏液和杯状细胞均减少(P<0.01,P<0.05),BALF中的SA、SA/Urea、MUC5AC和MPO水平降低(P<0.01),以及Urea水平升高(P<0.01),肺组织中的ROS和Objective To observe the effect of Jinshui Liujun decoction on airway lesions in mice with COPD and explore its possible mechanism.Methods 48 C57BL/6 mice were randomly divided into blank group,model group,Jinshui Liujun decoction group and NAC group,with 12 in each group.The COPD mouse model was established by intranasal drip of LPS and smoking,and the corresponding drugs were given intragastric administration for 14 days after the model was established.Observe the general condition of the mice,measure the MV,PEF and PIF of the mice with the small animal lung function instrument,semi quantitatively evaluate the inflammation of the lung tissue,the thickness of the alveolar septum and the thickness of the airway wall with HE staining,and observe the airway mucus secretion and goblet cell proliferation with PAS staining.The content of MPO,SA and Urea in BALF was detected by the kit,and the ratio of SA and Urea was calculated.The content of MUC5AC in BALF was detected by ELISA.The levels of ROS,GSH,GSSG and GR in lung tissue were detected with the kit,and the ratio of GSH and GSSG was calculated.The expression level of CFTR mRNA in lung tissue was detected by qRT-PCR.Western blot was used to detect the expression level of CFTR protein in lung tissue.Results Compared with the control group,the growth of mice in the model group was poor.The body weight at each time point during the modeling period decreased(P<0.01),and the indexes of MV,PEF and PIF decreased(P<0.01).The lung tissue pathological score,alveolar septal thickness,airway wall thickness,airway mucus and goblet cell increased(P<0.01).The levels of SA,SA/Urea,MUC5AC and MPO in BALF increased(P<0.01),and the level of Urea decreased(P<0.01),The levels of ROS and GSSG in lung tissue increased(P<0.01),and the levels of GSH,GSH/GSSG,and GR decreased(P<0.01).The expression levels of CFTR mRNA and protein in lung tissue decreased(P<0.01).Compared with the model group,the growth condition of COPD mice improved,the body weight increased at each time point during the m

关 键 词：金水六君煎 慢性阻塞性肺疾病 气道黏液 气道病变 氧化应激 CFTR获得性缺陷 N- 乙酰半胱氨酸 

分 类 号：R285.5[医药卫生—中药学]