麦门冬汤通过调节TLR4-NF-κB通路对大鼠肺纤维化作用机制研究  

Study on Mechanism of Maimendong Decoction(麦门冬汤)on pulmonary Fibrosis in Rats by Regulating TLR4-NF-κB Pathway

作　　者：黄健华 刘锐[2] 周怡菲 刘丽[1] 陈波[1] HUANG Jianhua;LIU Rui;ZHOU Yifei;LIU Li;CHEN Bo(Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China;Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学附属瑞康医院,广西南宁530011

出　　处：《辽宁中医杂志》2024年第11期188-193,I0007,I0008,共8页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金地区项目(81860829);广西自然科学基金项目(2020GXNSFAA297158);第五批全国临床优秀人才研修项目(国中医药人教函【2022】1号);广西青年岐黄学者培养项目(桂中医药科教发【2022】13号);广西研究生教育创新计划项目(YCSY2022032)。

摘　　要：目的通过博来霉素诱导的大鼠肺纤维化模型,使用麦门冬汤进行干预,观察各组大鼠肺组织病理学情况及各项指标的变化,研究麦门冬汤通过TLR4/NF-κB信号通路对肺纤维化大鼠的干预作用和机制。方法采购SPF级别SD雄性大鼠90只,将其随机分成6组,每组各15只,分别是空白组、模型组、强的松组、麦门冬汤低剂量组、麦门冬汤中剂量组和麦门冬汤高剂量组。干预后14天和28天,分别收集各组大鼠肺组织和血清样本,HE染色和Masson染色观察大鼠肺组织病理学变化,免疫组化法测定大鼠肺组织中Toll样受体4(Toll-like recepter 4,TLR4)、核因子激活的B细胞κ轻链增强(nuclear factor-κB,NF-κB)的表达情况,通过ELISA法测定大鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)的水平,实时荧光定量PCR测定大鼠肺组织中细胞角蛋白(Cytokeratin)、N-钙粘蛋白(N-cadherin)、TLR4、NF-κB的mRNA因表达水平。结果建模后14天,对比空白组,模型组HE和Masson染色显示大鼠肺组织病理异常,大鼠肺组织结构明显受损及构架明显紊乱,大量的炎症细胞浸润,肺泡壁增厚,蓝色胶原纤维明显增生,经药物干预后,对比模型组,各治疗组大鼠肺泡炎症及肺纤维化程度有所减轻;建模后28天,对比空白组,模型组HE和Masson染色显示大鼠肺组织结构重度受损以及构架严重紊乱,肺泡几乎被纤维团覆盖,有大量胶原沉积及蓝色胶原纤维增生,经药物干预后,对比模型组,各治疗组大鼠肺组织损伤及纤维化减轻。建模后14、28天,对比空白组,模型组中的TLR4、NF-κB、N-cadherin、TNF-α、IL-6表达水平均升高,Cytokeratin的表达水平降低;治疗后14、28天,对比模型组,强的松组和麦门冬汤各剂量组中的TLR4、NF-κB、N-cadherin、TNF-α、IL-6表达水平均降低,Cytokeratin表达水平升高。结论在博莱霉素诱导大鼠肺纤维化模型中,麦门冬汤能�Objective The effect of Maimendong Decoction(麦门冬汤)on pulmonary fibrosis induced by bleomycin in rats was studied.The changes of lung histopathology and various indexes of rats in each group were observed and the intervention effect and mechanism of Maimendong Decoction on pulmonary fibrosis rats were studied through Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods Ninety male SD rats with SPF grade were purchased and randomly divided into 6 groups with 15 rats in each group,which were blank group,model group,prednisone group,Maimendong Decoction low,medium and high dose groups.At 14 and 28 days after intervention,lung tissue and serum samples were collected from each group,and the histopathological changes of the lung were observed by HE staining and Masson staining.The expressions of TLR4 and NF-κB in the lung tissues of the rats were determined by immunohistochemistry.Serum levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were determined by ELISA,and gene expression levels of Cytokeratin,N-cadherin,TLR4 and NF-κB in rat lung tissue were determined by real-time quantitative PCR.Results 14 days after modeling,compared with the blank group,HE and Masson staining in the model group showed pathological abnormalities in rat lung tissue,significant structural damage and disorder of rat lung tissue,infiltration of a large number of inflammatory cells,thickening of alveolar walls,and significant proliferation of blue collagen fibers.After drug intervention,compared with the model group,the degree of alveolar inflammation and pulmonary fibrosis in each treatment group of rats was reduced.28 days after modeling,compared with the blank group,HE and Masson staining of the model group showed that the lung tissue structure of rats was severely damaged and the framework was seriously disturbed,and the alveoli were almost covered by fibrous masses,with a large amount of collagen deposition and blue collagen fiber hyperplasia.After drug intervention,the lung tissue damage a

关 键 词：肺纤维化 麦门冬汤 上皮-间质细胞转化 TLR4-NF-κB信号通路 细胞角蛋白 肿瘤坏死因子-Α 

分 类 号：R256.1[医药卫生—中医内科学]