灯盏花素对慢性阻塞性肺疾病模型大鼠气道重塑的影响

目的观察灯盏花素对慢性阻塞性肺疾病(COPD)模型大鼠气道重塑的影响,探讨灯盏花素防治COPD模型大鼠气道重塑的机制。方法将48只雄性Wistar大鼠随机分为空白组、模型组、灯盏花素组,每组16只,模型组、灯盏花素组采用熏烟加气管内滴入内毒素脂多糖的方法建立大鼠COPD模型,灯盏花素组予灯盏花素灌胃,空白组及模型组给予等容积0.9%氯化钠注射液灌胃。3组分别于给药第7、28 d各处死8只大鼠,行肺组织病理学观察,测定支气管厚度及支气管胶原纤维厚度,用免疫组化法测定肺组织中基质金属蛋白酶9(MMP-9)、转化生长因子β(TGF-β)蛋白阳性表达面积比率,用实时荧光定量聚合酶链式反应(PCR)法测定肺组织中Smad3和Smad7 mRNA水平。结果与空白组比较,模型组及灯盏花素组第7、28 d大鼠支气管壁厚度及胶原纤维厚度均明显升高(P0.01)。与模型组比较,灯盏花素组第7、28 d大鼠支气管壁厚度及胶原纤维厚度均下降(P0.05,P0.01)。灯盏花素组内7 d与28 d比较,支气管壁厚度无明显差异,28 d时支气管胶原纤维厚度下降(P0.05)。与空白组比较,模型组第7、28 d大鼠肺组织MMP-9、TGF-β表达增强,Smad 3 mRNA水平升高(P0.01),Smad 7 mRNA水平降低(P0.01);灯盏花素组第7 d肺组织MMP-9、TGF-β表达增强(P0.05),Smad3 mRNA水平升高(P0.01),Smad7 mRNA水平降低(P0.01)。与模型组比较,灯盏花素组第28 d大鼠肺组织MMP-9、TGF-β表达减少(P0.01,P0.05),Smad 3 mRNA水平降低(P0.01),Smad 7 mRNA水平升高(P0.01)。灯盏花素组内7 d与28 d比较,28 d时大鼠肺组织MMP-9、TGF-β及Smad3 mRNA水平降低(P0.05),Smad7 mRNA水平升高(P0.05)。结论灯盏花素可抑制COPD模型大鼠支气管壁厚度及胶原纤维厚度的增加,降低COPD大鼠肺组织中的MMP-9、TGF-β及Smad3 mRNA水平,升高Smad7 mRNA水平,从而可以延缓或改善COPD气道重塑的疾病进程。

Effects of Breviscapine on airway remodeling in rats with chronic obstructive pulmonary disease model

Objective To observe the effects of breviscapine on airway remodeling in rat with chronic obstructive pulmonary disease (COPD) model, and to explore the mechanism of breviscapine on preventing airway remodeling in rats with COPD model.Methods 48 male Wistar rats were randomly divided into blank group, model group and breviscapine group, 16 rats in each group.The rat model of COPD was established by cigarette smoking combined with intratracheal instillation of lipopolysaccharide(LPS) in model group and breviscapine group.Breviscapine group was treated by breviscapine for gavage, and the blank group was treated by equal volume of 0.9% sodium chloride injection.8 rats were executed at the 7th d and 28th d after administration in each group.The pathology observation in lung tissue was conducted, and the thickness of bronchi and bronchial collagen fibers were measured.The positive expression area ratio of matrix metalloproteinases 9 (MMP-9) and transforming growth factor β (TGF-β) in lung tissue were detected by immunohistochemical method, and the Smad3 mRNA and Smad7 mRNA were detected by real time fluorescent quantitative polymerase chain reaction (PCR).Results Compared with blank group, the thickness of bronchi wall and bronchial collagen fibers at the 7th d and 28th d significantly increased in model group and breviscapine group (P<0.01).Compared with model group, the thickness of bronchi wall and bronchial collagen fibers at the 7th d and 28th d reduced in breviscapine group (P<0.05, P<0.01).There was no statistical difference on the thickness of bronchi wall between the 7th d and 28th d in breviscapine group, and the thickness of bronchial collagen fibers at 28th d reduced (P<0.05).Compared with blank group, the expression of MMP-9 and TGF-β in lung tissue at the 7th d and 28th d increased in model group,and Smad3 mRNA increased (P<0.01), Smad7 mRNA decreased (P<0.01), and the expression of MMP-9 and TGF-β of lung tissue at the 7th d increased in breviscapine group(P<0.05), Smad3 mRNA increased (P<0.01), Smad7 mRNA decreased (P<0.01).Compared with model group, the expression of MMP-9 and TGF-β in lung tissue at the 28th d decreased in breviscapine group (P<0.01, P<0.05), Smad3 mRNA decreased (P<0.01), Smad7 mRNA increased (P<0.01).The expression of MMP-9 and TGF-β in lung tissue and Smad3 mRNA at the 28th d were lower than that at the 7th d in breviscapine group (P<0.05), and Smad7 mRNA higher (P<0.05).Conclusion Breviscapine can inhibit the increase of the thickness of bronchial wall and collagen fibers in COPD model rats, decrease the MMP-9, TGF-β and Smad3 mRNA in lung tissue, and increase Smad7 mRNA, thus can delay or improve the disease process of COPD airway remodeling.