水飞蓟宾对小鼠肺纤维化及氧化应激的作用及其机制  

Effect and mechanism of silybin on improving pulmonary fibrosis and oxidative stress in mice by mediating BMP9/SMAD signaling pathways

作　　者：任维鑫 王俊华 柳梅 韩成美 丁雷 Ren Wei-Xin;Wang Jun-Hua;Liu Mei;Han Cheng-Mei;Ding Lei(Department of Infection,the Affilicated People's Hospital of Shandong First Medical University/Jinan People's Hospital,Jinan,Shandong 271100,China;Department of Respiratory,the Affilicated People's Hospital of Shandong First Medical University/Jinan People's Hospital,Jinan,Shandong 271100,China)

机构地区：[1]山东第一医科大学附属人民医院/济南市人民医院感染科,山东济南271100 [2]山东第一医科大学附属人民医院/济南市人民医院呼吸内科,山东济南271100

出　　处：《解放军医学杂志》2022年第12期1201-1208,共8页Medical Journal of Chinese People's Liberation Army

基　　金：山东省市级科技项目(WSCG2018-007)。

摘　　要：目的探讨水飞蓟宾对小鼠肺纤维化和氧化应激的缓解作用及其可能机制。方法120只小鼠随机分为假手术组、肺损伤模型组、地塞米松组,以及水飞蓟宾低、中、高剂量组,每组20只。采用博来霉素诱导小鼠肺损伤模型,地塞米松组及水飞蓟宾低、中、高剂量组分别灌胃给予地塞米松(3 mg/kg)和水飞蓟宾(120、240、480 mg/kg),假手术组和肺损伤模型组灌胃给予等量生理盐水,连续28 d。收集各组支气管肺泡灌洗液(BALF)和肺组织,镜检计数BALF中炎性细胞数,HE染色观察肺组织病理变化,干湿重法测定肺湿/干重比(W/D),ELISA法检测肺组织中羟脯氨酸(HYP)含量以及BALF中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、总抗氧化能力(T-AOC)、丙二醛(MDA)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β水平,Western blotting检测肺组织中骨形态发生蛋白9/骨成型蛋白受体2(BMP9/BMPR2)/SMAD信号通路相关蛋白的表达。结果与假手术组比较,肺损伤模型组小鼠BALF炎性细胞数、肺损伤评分、肺纤维化评分、肺W/D、HYP含量,以及BALF中MDA、TNF-α、IL-1β、IL-6水平明显升高,BALF中SOD、GSH-Px活性和T-AOC水平,以及肺组织中BMP9、BMPR2、p-SMAD1/5/9蛋白相对表达水平明显降低(P<0.05);与肺损伤模型组比较,水飞蓟宾高剂量组及地塞米松组小鼠BALF炎性细胞数、肺损伤评分、肺纤维化评分、肺W/D、HYP含量,以及BALF中MDA、TNF-α、IL-1β、IL-6水平明显降低,BALF中SOD、GSH-Px活性和T-AOC水平,以及肺组织中BMP9、BMPR2、p-SMAD1/5/9蛋白相对表达水平明显升高(P<0.05)。结论水飞蓟宾可能通过介导BMP9/SMAD信号通路,缓解博来霉素诱导的肺损伤小鼠肺纤维化和氧化应激,从而实现肺损伤修复。Objective To explore the effect and mechanism of silybin on improving pulmonary fibrosis and oxidative stress in mice.Methods A total of 120 mice were randomly divided into sham operation group,lung injury model group,dexamethasone group,low-dose,medium-dose and high-dose silybin groups(20 mice each).The models of pulmonary injury mice were induced by bleomycin.Mice in dexamethasone group,low-,medium-and high-dose silybin groups were given intragastric administration of dexamethasone(3 mg/kg)and silybin(120,240,480 mg/kg),while sham operation group and lung injury model group were given the same volume of normal saline for 28 d.The bronchoalveolar lavage fluid(BALF)and pulmonary tissues of each group were collected.The count of inflammatory cells in BALF was examined by microscopy.The pathological damage of pulmonary tissues was observed by HE staining.The pulmonary wet/dry weight ratio(W/D)was detected by wet-dry weighting method.The content of hydroxyproline(HYP)in pulmonary tissues,and levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total antioxidant capacity(T-AOC),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1βin BALF were detected by ELISA.The expressions of bone morphogenetic protein 9(BMP9)/bone morphogenetic protein receptor 2(BMPR2)/SMAD signaling pathways-related proteins were detected by Western blotting.Results Compared with sham operation group,count of BALF inflammatory cells,scores of pulmonary injury and fibrosis,W/D,HYP,and MDA,TNF-α,IL-1βand IL-6 levels in BALF increased significantly,while SOD,GSH-Px activity,T-AOC levels in BALF,as well as the relative expression levels of BMP9,BMPR2 and p-SMAD1/5/9 in lung tissues decreased significantly in lung injury model group(P<0.05).Compared with lung injury model group,count of BALF inflammatory cells,scores of pulmonary injury and fibrosis,W/D,HYP,and MDA,TNF-α,IL-1βand IL-6 levels in BALF decreased significantly,while SOD,GSH-Px activity,T-AOC level in BALF,and BMP9,BMPR2 and p-SMAD1/5/9 exp

关 键 词：水飞蓟宾 博来霉素 谷胱甘肽过氧化物酶 超氧化物歧化酶 丙二醛 总抗氧化能力 肿瘤坏死因子-α 白细胞介素 

分 类 号：R932[医药卫生—生药学]