摘要
目的观察益气活血化痰方通过调控白细胞介素8(IL-8)/黏蛋白5ac(MUC5ac)信号通路对慢性阻塞性肺疾病(COPD)大鼠气道重塑及平滑肌厚度的影响。方法选取53只SPF级SD雄性大鼠,随机抽取10只作为健康组,剩余43只大鼠建立COPD模型,40只采用数字随机表法分为模型组、中药组、抑制剂组、联合组,每组10只。中药组大鼠给予益气活血化痰方32 g/kg灌胃,抑制剂组大鼠予腹腔注射吡咯烷二硫代甲酸铵(PDTC)100 mg/kg,联合组予益气活血化痰方灌胃后腹腔注射PDTC 100 mg/kg。健康组、模型组正常饲养,不予干预。采用动物肺功能检测仪测定各组大鼠肺功能[用力肺活量(FVC)、呼气峰值流速(PEF)、第1 s用力呼气容积(FEV 1)],酶联免疫吸附法检测肺泡灌洗液中IL-8、IL-17、肿瘤坏死因子α(TNF-α)水平,蛋白免疫印迹法检测肺组织IKB激酶2(IKK2)、磷酸化IKB激酶α(p-IKBα)、核转录因子κB(NF-κB)、MUC5ac蛋白表达,苏木素-伊红(HE)染色观察肺组织病理形态。结果与健康组比较,模型组、中药组、抑制剂组、联合组FVC、PEF、FEV 1水平均降低(P<0.05);与模型组比较,中药组、抑制剂组、联合组FVC、PEF、FEV 1水平均升高(P<0.05);联合组FVC、PEF、FEV 1水平均高于中药组、抑制剂组(P<0.05)。与健康组比较,模型组、中药组、抑制剂组、联合组肺泡灌洗液IL-8、IL-17、TNF-α水平均升高(P<0.05);与模型组比较,中药组、抑制剂组、联合组肺泡灌洗液IL-8、IL-17、TNF-α水平均降低(P<0.05);联合组肺泡灌洗液IL-8、IL-17、TNF-α水平均低于中药组、抑制剂组(P<0.05)。与健康组比较,模型组、中药组、抑制剂组、联合组支气管平滑肌厚度均增加(P<0.05);与模型组比较,中药组、抑制剂组、联合组支气管平滑肌厚度均降低(P<0.05);联合组支气管平滑肌厚度低于中药组、抑制剂组(P<0.05)。与健康组比较,模型组、中药组、抑制剂组、联合组肺组织IKK2、p-IKBα、NF-κB、MUC5ac表达均升高(P<0.05);与模型组比较,中药组、抑制剂组、联合组肺组织IKK2、p-IKBα、NF-κB、MUC5ac表达均降低(P<0.05);联合组肺组织IKK2、p-IKBα、NF-κB、MUC5ac表达均低于中药组、抑制剂组(P<0.05)。结论益气活血化痰方通过抑制IL-8/MUC5ac通路活化,抑制炎性反应,并降低气管平滑肌厚度,抑制气道重塑,改善肺功能。
Objective To observe the effects of Yiqi Huoxue Huatan Prescription on immune disorders and smooth muscle thickness in rats with chronic obstructive pulmonary disease(COPD)via regulating interleukin-8(IL-8)/mucins 5ac(MUC5ac)signaling pathway.Methods A total of 53 SPF-grade SD male rats were selected in this study,10 rats were randomly selected as the healthy group,and the remaining 43 rats were used to establish a COPD model.In modeling,3 rats died unexpectedly,and the remaining 40 rats were divided into the model group,the traditional Chinese medicine(TCM)group,the inhibitor group and the combination group by digital random table method,with 10 rats in each group.Rats in the TCM group were given 10 mL/kg Yiqi Huoxue Huatan Prescription(dose 3.2 g/mL)by intragastric administration,rats in the inhibitor group were intraperitoneally injected with 100 mg/kg ammonium pyrrolidine dithiocarbamate(PDTC),and rats in the combination group were intraperitoneally injected with 100 mg/kg PDTC after intragastric Yiqi Huoxue Huatan Prescription.Healthy group and model group were given equal volume 0.9%sodium chloride injection without intervention.Lung function of rats in each group was measured using an animal lung function detector,including:forced vital capacity(FVC),peak expiratory flow rate(PEF)and forced expiratory volume(FEV 1).The levels of IL-8,IL-17 and tumor necrosis factor alpha(TNF-α)in alveolar lavage fluid were measured by enzyme-linked immunosorbent assay.The expressions of IKB kinase 2(IKK2),phosphor-IKB alpha(p-IKBα),nuclear factor kappa B(NF-κB)and MUC5ac in lung tissues were detected by western blot.Hematoxylin eosin(HE)staining was used to observe the pathological morphology of lung tissues.Results Compared with healthy group,the levels of FVC,PEF and FEV 1 in model group,TCM group,inhibitor group and combination group decreased significantly(P<0.05).Compared with model group,the levels of FVC,PEF,and FEV 1 in TCM group,inhibitor group,and combination group increased obviously(P<0.05).The levels of FVC,PEF,and FEV 1 in the combination group were remarkably higher than those in the TCM group and inhibitor group(P<0.05).Compared with the healthy group,the model group,TCM group,inhibitor group,and combination group had significantly higher levels of IL-8,IL-17,and TNF-αin alveolar lavage fluid(P<0.05).Compared with the model group,the TCM group,inhibitor group,and combination group had significantly lower levels of IL-8,IL-17,and TNF-αin alveolar lavage fluid(P<0.05).The levels of IL-8,IL-17,TNF-αin the combination group were remarkably lower than those in the TCM group and inhibitor group(P<0.05).Compared with the healthy group,the model group,TCM group,inhibitor group and combination group had significantly increased thickness of bronchial smooth muscle(P<0.05).Compared with the model group,the thickness of bronchial smooth muscle in the TCM group,inhibitor group,and combination group decreased remarkably(P<0.05).The thickness of bronchial smooth muscle in the combination group was obviously lower than that in the TCM group and inhibitor group(P<0.05).Compared with the healthy group,the model group,TCM group,inhibitor group and combination group had significantly up-regulated levels of IKK2,p-IKBα,NF-κB and MUC5ac in lung tissues(P<0.05).Compared with the model group,the TCM group,inhibitor group,and combination group showed significantly down-regulated levels of IKK2,p-IKBα,NF-κB and MUC5ac in lung tissues(P<0.05).The levels of IKK2,p-IKBα,NF-κB and MUC5ac in the combination group were significantly lower than those in the TCM group and inhibitor group(P<0.05).Conclusion Yiqi Huoxue Huatan Prescription inhibits the activation of IL-8/MUC5ac pathway,inhibits inflammatory response,reduces the thickness of tracheal smooth muscle,inhibits airway remodeling and improves lung function.
作者
陈慧婷
王盛隆
白丽
CHEN Huiting;WANG Shenglong;BAI Li(Department of Pulmonology,Shanxi Provincial Integrated TCM and WM Hospital,Taiyuan,Shanxi 030013)
出处
《河北中医》
2023年第8期1335-1340,共6页
Hebei Journal of Traditional Chinese Medicine
基金
山西省基础研究计划自由探索类自然科学研究面上项目(编号:20210302123342)。
关键词
肺疾病
慢性阻塞性
大鼠
动物模型
益气活血化痰
免疫
平滑肌
Chronic obstructive pulmonary disease(COPD)
Rats
Animal model
Yiqi Huoxue Huatan Prescription
Immunization
Smooth muscle
