金水尘肺方抑制TGF-β1/Smad3活化改善矽肺大鼠肺纤维化  

Jinshui Chenfei formula attenuates silica-induced pulmonary fibrosis in rats via inhibi-tion of TGF-β1/Smad3 activation

作　　者：杨帆[1,2] 侯润苏 闫鑫华 白云苹[1,2,3] 田燕歌[1,2,3] 赵鹏 YANG Fan;HOU Run-su;YAN Xin-hua;BAI Yun-ping;TIAN Yan-ge;ZHAO Peng(Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Henan University of Chinese Medicine,Zhengzhou,Henan 450046,China;Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province&Education Ministry of P.R.China,Henan University of Chinese Medicine,Zhengzhou,Henan 450046,China;Academy of Chinese Medical Sciences,Henan University of Chinese Medicine,Zhengzhou,Henan 450046,China)

机构地区：[1]河南中医药大学,河南省中医药防治呼吸病重点实验室,河南郑州450046 [2]河南中医药大学,呼吸疾病中医药防治省部共建协同创新中心,河南郑州450046 [3]河南中医药大学中医药科学院,河南郑州450046

出　　处：《时珍国医国药》2023年第6期1285-1289,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81904170);河南省重点研发与推广专项(科技攻关)(212102310356);河南省中医药科学研究专项课题(20-21ZYZD01)。

摘　　要：目的基于TGF-β1/Smad3通路探讨金水尘肺方改善SiO_(2)诱导矽肺模型大鼠肺部病变作用及其机制。方法32只SPF级雄性SD大鼠随机分为正常组、模型组、汉防己甲素组(27 mg·kg^(-1)·d^(-1))、金水尘肺方组(9.72 g·kg^(-1)·d^(-1))。采用一次性非暴露式气管滴注二氧化硅(SiO_(2))混悬液(250 mg·kg^(-1))制备矽肺模型大鼠;于造模后第15~28天给予相应药物灌胃。第29天检测大鼠肺功能用力肺活量(FVC)、最大呼气中期流量(MMEF)、静态肺顺应性(Cchord)以及动态肺顺应性(Cydn)变化。HE染色和Masson染色观察肺组织病理改变;酶联免疫吸附实验检测肺组织肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、IL-6水平;免疫荧光组织化学法检测肺组织CD45、Ⅰ型胶原蛋白(COLⅠ)表达;免疫组织化学法检测COLⅠ、α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)和p-Smad3表达;蛋白免疫印迹法检测α-SMA、TGF-β1、纤维连接蛋白(FN)和p-Smad3蛋白表达。结果与正常组比较,模型组大鼠肺泡断裂融合,纤维结节包裹SiO_(2)颗粒呈肉芽肿改变;肺功能FVC、MMEF、Cchord和Cydn显著降低(P<0.01),肺组织TNF-α、IL-1β、IL-6水平显著升高(P<0.05,P<0.01),肺组织CD45、COLⅠ双阳性细胞增多,肺组织COLⅠ、α-SMA、FN、TGF-β1、p-Smad3蛋白表达显著升高(P<0.05,P<0.01)。与模型组比较,金水尘肺方组和汉防己甲素组肺组织病理形态有所改善,肺功能FVC、MMEF、Cchord和Cydn显著升高(P<0.05,P<0.01),肺组织TNF-α、IL-1β、IL-6水平显著降低(P<0.05,P<0.01),肺组织CD45、COLⅠ双阳性细胞减少,肺组织COLⅠ、α-SMA、FN、TGF-β1、p-Smad3蛋白表达显著降低(P<0.05,P<0.01)。其中金水尘肺方组Cydn较汉防己甲素组显著升高(P<0.01),TGF-β1、p-Smad3的表达较汉防己甲素组显著降低(P<0.01)。结论金水尘肺方可以改善SiO_(2)诱导的矽肺模型大鼠肺纤维化,其机制可能与抑制TGF-β1/Smad3通路有关。Objective To explore the effect of Jinshui Chenfei Formula(JCF)on improving SiO_(2)-induced silicosis in rats and its undrlying mechanisms by TGF-β1/Smad3 pathway.Methods 32 SPF male SD rats were randomly divided into control group,model group,tetrandrine group(27 mg·kg^(-1)·d^(-1)),and JCF group(9.72 g·kg^(-1)·d^(-1)).The experimental silicosis model was established by intratracheal injection with SiO_(2) suspension(250 mg·kg^(-1)).From day 15 to 28,silicosis rats were treated with tetrandrine or JCF.On day 29,the changes of pulmonary function,including FVC,MMEF,Cchord and Cydn were detected.Then,all rats were sacrificed,and the lung tissues were collected.The pathological changes of lung tissue were ana-lyzed by HE and Masson staining.The levels of TNF-α,IL-6 and IL-1βin lung tissue were detected by enzyme-linked im-munosorbent assay.The expression levels of CD45 and COLⅠin lung tissue was detected by immunofluorescence.The protein levels of COLⅠ,α-SMA,TGF-β1 and p-Smad3 expressions were detected by immunohistochemistry.The protein expres-sions ofα-SMA,FN,TGF-β1,p-Smad3 were detected by Western blot.Results Compared with those in the control group,pathological changes such as alveolar structure destruction,the fibrous nodules encapsulated SiO_(2) particles and granulomatous changes,were increased in lung tissues of model group;pulmonary function including FVC,MMEF,Cchord and Cydn were sig-nificantly decreased(P<0.01);the levels of TNF-α,IL-1βand IL-6 were significantly increased(P<0.05,P<0.01);the CD45 and COLI double-staining cells in the lung tissue were significantly increased,and the protein expressions of COLI,α-SMA,TGF-β1 and p-Smad3 in the lung tissue were significantly increased(P<0.05,P<0.01).Compared with the mod-el group,the pathological changes of the lung tissue were suppressed;the pulmonary function FVC,MMEF,Cchord and Cydn were significantly increased(P<0.05,P<0.01);and the levels of TNF-α,IL-1βand IL-6 were significantly decreased(P<0.05,P<0.01);the CD45 and COLI double-sta

关 键 词：矽肺 纤维化 金水尘肺方 TGF-β1/Smad3 

分 类 号：R285.5[医药卫生—中药学]