灯盏花素对低氧诱导的大鼠肺泡炎症及细胞外基质沉积的作用  

Effect of Breviscapine on hypoxia-induced inflammation and ECM deposition in SD Rats' lung

作　　者：张旭[1] 魏华华 谢敏[3] 王永生[1] ZHANG Xu;WEI Huahua;XIE Min;WANG Yongsheng(Department of Respiratory Medicine,Chengdu First People＇s Hospital,Chengdu 610041,China;Department of Respiratory Medicine,The Third People＇s Hospital of Chengdu,Chengdu 610031,China;Department of Respiratory and Critical Care Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]成都市第一人民医院呼吸内科,四川成都610041 [2]成都市第三人民医院呼吸内科,四川成都610031 [3]四川大学华西医院呼吸与危重症医学科,四川成都610041

出　　处：《西部医学》2018年第8期1099-1106,共8页Medical Journal of West China

基　　金：四川省科技厅攻关项目(0040205301469)

摘　　要：目的观察间歇性常压性低氧对大鼠肺组织中TGF-β1、Smad4蛋白、Col I、TNF-α表达量的影响及灯盏花素对其的干预作用。方法将80只SPF级SD大鼠随机分为对照组、单纯低氧组、低氧+低剂量灯盏花素组,低氧+高剂量灯盏花素组,分别置于常氧或间歇性常压性低氧(101kPa,10%O2,每天低氧8h)条件下,于3、7、14、21d每组随机处死5只大鼠。HE染色观察肺组织病理改变;免疫组化染色检测肺组织TGF-β1、Col I表达;Western-blot法检测Smad4通道蛋白的表达;RT-PCR检测TGF-β1、Col I的mRNA表达量;ELISA法检测肺泡灌洗液(BALF)中TNF-α的表达。结果 HE染色结果显示低氧可引起大鼠肺组织水肿及炎症细胞浸润,且随低氧时间延长,炎症反应加重,肺泡间隔逐渐增厚,而药物干预组水肿、炎症反应及间隔增厚程度均较单纯低氧组轻;单纯低氧组大鼠肺组织中TGF-β1、Smad4、Col I蛋白及TGF-β1mRNA、Col I mRNA表达量均较对照组增高(P<0.01),随低氧时间延长,各因子表达量逐渐增高,且Smad4蛋白表达水平与TGF-β1表达水平呈正相关(r=0.944,P<0.01),BALF中TNF-α的表达上调(P<0.01)。药物干预组TGF-β1、Smad4、Col I及TNF-α蛋白、Col I mRNA的表达量均较单纯低氧组下调(P<0.05),且低氧+高剂量灯盏花素组较低氧+低剂量灯盏花素组降低(P<0.05),但TGF-β1mRNA的表达量单纯低氧、低氧+低剂量灯盏花素、低氧+高剂量灯盏花素三组比较差异无统计学意义(P>0.05)。结论低氧可能通过诱发肺泡水肿及炎症反应,上调TNF-α水平、激活TGF-β1/Smads通路从而增加Col I合成,引发细胞外基质沉积;灯盏花素则通过调节TGF-β1及Smad4蛋白表达,下调Col ImRNA及蛋白表达,并降低TNF-α水平,抑制低氧诱导的肺组织结构改变。Objective To observe the effect of intermittent normobaric hypoxia on the expression of Transforming Growth Factor beta1（TGF-β1）,Smad4,collagen I（Col I）,Tumor Necrosis Factor alpha（TNF-α）and on the changes in the histological structure of the Rats＇ Lung and the effect of breviscapine on the hypoxia-induced changes.Methods 80 Rats were randomly divided into control group（group A）,hypoxia group（group B）,hypoxia and low-dose breviscapine group（group C）,hypoxia and high-dose breviscapine group（group D）.They were placed in normal environment or intermittent normobaric hypoxia（101 kpa,10% O2,8 hevery day）,respectively.At the 3 ^d,7^ th,14 ^th,21 ^st day,5 rats from each group were killed,Hematoxylin and Eosin staining（HE staining）was taken to observe the pathological change in the rats＇ lung,Immunohistochemical staining to detect the protein level of TGF-β1,Col I and Western blot to detect the expression of Smad4.RT-PCR was performed to detect the mRNA levels of TGF-β1 and Col I.Besides,the TNF-αin the bronchoalveolar lavage fluid（BALF）were determined by ELISA.Results HE staining shown that mild edema and inflammatory cell infiltration appear in the lung tissue at the 3 ^d day,meanwhile inflammation gradually aggravated,and the interalveolar septum became thicker and thicker as the time of exposing to hypoxia going on.In both breviscapine groups,the degree of hypoxia-induced inflammation and the septum changes were lighter than that in the hypoxia group.Compared with the normoxia group,not only the protein expression of TGF-β1,Smad4,Col I and TNF-α,but also the mRNA expression of TGF-β1 and Col I were elevated,and with longer exposure to hypoxia they increase further.It also demonstrated positive correlation between the protein level of TGF-β1 and Smad4.Meanwhile the protein of TGF-β1,Smad4,Col I,TNF-αand Col I mRNA detected in both the breviscapine groups were declined.The expression level in group D was lower than group C.There is no statistical difference in TGF

关 键 词：低氧 肺纤维化 TGF-β1 Ⅰ型胶原 SMADS SMAD4 TNF-α 

分 类 号：R-332[医药卫生]