二陈汤加味对慢性阻塞性肺疾病大鼠肺组织沉默信息调节因子1、内皮素-1表达的影响

Effect of Er Chen decoction on the expression of Sirt1 and endothelin-1 in lung tissue of chronic obstructive pulmonary disease rat model

目的探索二陈汤加味对慢性阻塞性肺疾病(COPD)大鼠肺组织沉默信息调节因子(Sirt)1、内皮素(ET)-1表达的影响,分析其药效机制。方法将50只SD大鼠随机分正常组、模型组、二陈汤加味低、中、高剂量组〔10,20,40 g/(kg·d)〕,运用气管内滴注脂多糖(LPS)及烟熏法制备COPD大鼠模型,造模完成后灌胃生理盐水、二陈汤加味方治疗,使用潮气量(TV)、呼气流量峰(PEF)等来测定肺功能变化,免疫组织化学法(IHC)及实时荧光定量聚合酶链式反应(qPCR)检测其肺组织内Sirt1、ET-1基因及蛋白表达。结果与正常组比较,模型组TV、PEF含量均显著减小(P<0.01),肺组织中Sirt1蛋白及基因表达显著减弱(P<0.01),肺组织中ET-1蛋白及基因表达显著增强(P<0.01);与模型组相比,各治疗组PEF、TV均显著增加(P<0.05),各治疗组肺组织Sirt1蛋白及mRNA表达均显著升高(P<0.01)、ET-1蛋白及基因表达均显著降低(P<0.01);其中,二陈汤加味中剂量组肺功能PEF、VT显著升高(P<0.05),Sirt1 mRNA及蛋白表达明显升高(P<0.01)、ET-1蛋白及基因表达显著降低(P<0.001)。结论二陈汤加味能够显著对COPD大鼠肺功能和气流受限起到保护作用,通过升高肺组织Sirt1 mRNA和蛋白表达,减弱ET-1 mRNA和蛋白表达,对COPD大鼠炎症反应起到抑制作用,阻止肺组织血管重构。

Objective To investigate the effects of Er Chen decoction on the expression of silent information regulatory factor(Sirt)1 and endothelin(ET)-1 in lung tissue of chronic obstructive pulmonary diseases(COPD)rat model and provide experimental evidence for the pharmacodynamic mechanism of Er Chen decoction in the treatment of COPD.Methods 50 SD rats were randomly divided into control model,low〔10 g/(kg·d)〕,medium〔20 g/(kg·d)〕and high〔40 g/(kg·d)〕dose of Er Chen decoction groups.COPD model was made by intratracheal instillation of lipopolysaccharide(LPS)and fumigation.After the model was made,the rats were given normal saline and Er Chen decoction.Lung function was determined by using peak expiratory flow(PEF),tidal volume(TV)and other indexes,immunohistochemical(IHC)and real-time fluorescence quantitative PCR were used to detect the expressions of Sirt1,ET-1 gene and protein in lung tissue.Results Compared with the control group,the PEF and TV in the model group were significantly decreased(P<0.05),the protein and gene expressions of Sirt1 in lung tissue were significantly decreased(P<0.01),and the expression of protein and gene expressions of ET-1 in lung tissue were significantly increased in the model group(P<0.01).The PEF and TV increased,the protein and gene expressions of Sirt1 in lung tissue increased and the protein and gene expressions of ET-1 in lung tissue decreased in some degree in all treatment groups(P<0.05,P<0.01).Conclusions Er Chen decoction could effectively improve airflow limitation and lung function in COPD rats.It could inhibit inflammation reaction and prevent pulmonary vascular remodeling by increasing protein and gene expressions of Sirt1,inhibiting the protein and gene expressions of ET-1.