摘要
目的:探究壮药龙盘止咳方减轻急性支气管炎小鼠肺损伤的潜在机制。方法:KM小鼠采用烟熏法建立急性支气管炎模型,将造模成功的60只小鼠随机分为模型组、SB203580(p38 MAPK特异性抑制剂,5 mg/kg)组、高剂量(10.4 g/kg)龙盘止咳方组、低剂量(5.2 g/kg)龙盘止咳方组和龙盘止咳方(10.4 g/kg)+SB203580(5 mg/kg)组,每组12只,另将正常饲养的12只小鼠设为对照组。分组完成后进行药物干预,每天1次,连续1周。观察给药前后各组小鼠的一般状态变化;收集支气管肺泡灌洗液(BALF),ELISA法检测BALF中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-18水平;HE染色观察肺组织病理学变化;免疫组化法检测肺组织核因子κB(NF-κB)p65的阳性表达;RT-qPCR检测肺组织核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、含caspase募集结构域的凋亡相关斑点样蛋白(ASC)、caspase-1和IL-1β的mRNA表达水平;Western blot检测肺组织p38丝裂原活化蛋白激酶(MAPK)/NF-κB/NLRP3通路相关蛋白的表达。结果:与对照组相比,模型组小鼠出现咳嗽、流涕、呼吸急促等症状,且肺部支气管可见炎性细胞浸润、肺泡壁和支气管壁增厚、黏膜充血和水肿等病理损伤,肺组织NF-κB p65在胞核中呈强阳性表达,BALF中TNF-α、IL-1β和IL-18水平,肺组织NLRP3、IL-1β、ASC和caspase-1的mRNA表达水平,以及p-p38 MAPK/p38 MAPK、胞核NF-κB p65、NLRP3和cleaved caspase-1蛋白水平显著升高(P<0.05),胞质NF-κB p65蛋白水平显著降低(P<0.05)。与模型组相比,高、低剂量龙盘止咳方组和SB203580组小鼠咳嗽、流涕、呼吸急促等症状,以及肺部支气管炎症细胞浸润等病理损伤明显减轻,肺组织胞核NF-κB p65呈阳性表达的细胞数减少,BALF中TNF-α、IL-1β和IL-18水平,肺组织NLRP3、IL-1β、ASC和caspase-1的mRNA表达水平,以及p-p38 MAPK/p38 MAPK、胞核NF-κB p65、NLRP3和cleaved caspase-1蛋白水平显著降低(P<0.05),胞质NF-κB p65蛋
AIM:To explore the potential mechanism of Zhuang medicine Longpan-Zhike decoction reducing lung injury in mice with acute bronchitis.METHODS:Sixty-four KM mice were smoked to establish an acute bronchitis model,and 60 mice with successful modeling were randomly divided into model group,p38 MAPK specific inhibitor SB203580(5 mg/kg)group,high-dose(10. 4 g/kg)and low-dose(5. 2 g/kg)Longpan-Zhike decoction groups,and Longpan-Zhike decoction(10. 4 g/kg)+SB203580(5 mg/kg)group,with 12 mice in each group. The mice raised normally(n=12)served as control group. Drug intervention was performed after grouping,once a day for 1 week continuously. The changes in the general state of the mice in each group before and after the administration were observed. Bronchoalveolar lavage fluid(BALF)was collected,and the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-18 in BALF were measured by ELISA. The pathological changes of lung tissues were observed by HE staining. The positive expression of nuclear factor κB(NF-κB)p65 in lung tissues was observed by immunohistochemical method. The mRNA expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1 and IL-1β in lung tissues was detected by RT-qPCR.The expression of p38 mitogen-activated protein kinase(MAPK)/NF-κB/NLRP3 signaling pathway-related proteins in lung tissues was determined by Western blot.RESULTS:Compared with control group,the mice in model group showed symptoms such as cough,runny nose and shortness of breath,and pathological damage such as inflammatory cell infiltration,alveolar and bronchial wall thickening,and mucosal congestion and edema were seen in the lung bronchus. Strongly positive expression of NF-κB p65 in the nucleus was observed in the lung tissue,and the levels of TNF-α,IL-1β and IL-18 in the BALF,the mRNA expression of NLRP3,IL-1β,ASC and caspase-1 in the lung tissue,and the protein levels of p-p38 MAPK/
作者
韦杏
邹敏
李崇进
农志飞
蓝毓营
WEI Xing;ZOU Min;LI Chong-jin;NONG Zhi-fei;LAN Yu-ying(Zhuang Medical College,Guangxi University of Chinese Medicine,Nanning 530001,China;Department of Pediatrics,Guangxi International Zhuang Medicine Hospital,Nanning 530200,China;Department of Pediatrics,Maoming Hospital of Traditional Chinese Medicine,Maoming 525000,China;Department of Pediatrics,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第10期1828-1837,共10页
Chinese Journal of Pathophysiology
基金
科技部国家重点研发计划“中医药现代化研究重点专项”(No.2017YFC1703903)
广西壮族自治区教育厅高校中青年教师基础能力提升项目(No.2018KY0302)
广西壮族自治区中医药管理局自筹经费科研课题(No.GZZC2020114)
广西中医药大学一流学科建设开放课题(No.2019XK044)
广西中医药大学校级青年基金项目(No.2018QN034)。
