紫金牛、铁扫帚对COPD大鼠气道DCN、Smad7的干预研究

Study on the Intervention of Ardisia japonica and Lespedeza cuneata on Airway DCN Smad7 of COPD Rats

目的:探讨苗药紫金牛、铁扫帚通过对COPD大鼠模型DCN、Smad7、TGF-β1的干预抑制气道重塑的可能作用机制,并观察不同剂量苗药对COPD大鼠的疗效差异,以寻求最佳的量效关系。方法:将60只健康雄性SD大鼠随机分为健康对照组12只,COPD模型组48只,模型组利用熏香烟加气管内注入脂多糖的方法制作COPD大鼠模型,将模型组大鼠随机分为4组:COPD空白组、苗药高剂量组、苗药中剂量组和苗药低剂量组,每组各12只,造模结束后每组随机选取2只大鼠,同时选2只健康大鼠,行病理切片检查(苏木精–伊红染色,HE染色)做模型鉴定。对各苗药组分别对应给予高、中、低剂量苗药紫金牛、铁扫帚灌胃干预治疗4周,健康对照组及COPD空白组分别予灭菌生理盐水灌胃干预治疗4周。运用HE染色观察治疗后各组大鼠肺组织病理改变;用蛋白质印迹法(Western Blot)检测治疗后各组大鼠肺组织中DCN、Smad7、TGF-β1的表达;通过Pearson相关系数分析DCN、Smad7、TGF-β1蛋白之间的相关性。结果:烟熏加气管内注射脂多糖造模法使得实验大鼠出现与COPD相应的病理改变。各苗药组肺组织形态较COPD空白组有所改善,尤以苗药高剂量组最为明显。经苗药治疗后,COPD大鼠模型症状均有所缓解,尤以苗药高剂量组缓解最为明显。经苗药紫金牛、铁扫帚干预治疗后,Smad7、TGF-β1蛋白表达苗药各组与COPD空白组之间均有明显差异(P 0.05);苗药高、中剂量组之间Smad7、DCN、TGF-β1蛋白表达均无明显差异(P >0.05),但苗药高剂量组Smad7、DCN蛋白表达均数高于苗药中剂量组、TGF-β1蛋白表达均数低于苗药中剂量组。COPD大鼠模型肺组织中Smad7与DCN呈正性相关(P <0.01),二者与TGF-β1均呈负性相关(P <0.01)。结论:在COPD大鼠模型组肺组织中,TGF-β1含量显著升高,DCN、Smad7显著降低;经苗药紫金牛、铁扫帚干预治疗后,苗药治疗组TGF-β1明显降低,DCN、Smad7明显升高,综合比较后发现尤以苗药高剂量组变化最为明显,说明苗药高剂量组疗效最佳。TGF-β1与Smad7、DCN均呈负性相关,Smad7与DCN呈正性相关。

Objective:To discuss the mechanism of action of miao medicine(Ardisia japonica,Lespedeza cuneata)on inhibiting COPD rats'airway obstruction,through DCN,Smad7 and TGF-β1 intervention.To observe the treatment,different doses of miao medicine were used to treat the COPD rats,to find the best relationship of dose effect.Methods:The 60 healthy male SD rats were divided into normal group(12 rats)and COPD models group(48 rats)randomly.The COPD rat model was made by using the method of smoking cigarette and injecting LPS into the trachea.Later,the 48 rats of modeling were divided into 4 groups:blank group,high-miao medicine group,middle-miao medicine group and low-miao medicine group,12 rats in each group.8 rats belonging to COPD model(each group of two)and 2 healthy rats were randomly chosen for chip check to verify the models.Rats of miao medicine groups were treated with miao medicine of different doses;rats of normal group and blank group were treated with normal saline for four weeks,respectively.Using hematoxylin-eosin staining(HE staining)to observe rats’lungs pathological manifestations,western blot was used to detect the expression quantity of TGF-β1,Smad7 and DCN in rats’lungs,and Pearson correlation coefficient was used to analyze the relationship among proteins of TGF-β1,Smad7 and DCN.Results:Groups of HE staining results:in the lungs of COPD models group,rats had the same pathologic changes with COPD.The lungs of each miao medicine group rates had improved more than the blank group,especially the high-miao medicine group.Comparison among groups symptoms:the symptoms of each miao medicine group improved,especially the high-miao medicine group changed most.After miao medicine treatment,all miao medicine groups’protein expression of Smad7,TGF-β1 had obvious difference with the blank group(P<0.05);middle-miao and high-miao medicine groups'protein expression of DCN had obvious difference with the blank group(P<0.05).Low-miao medicine group had no obvious difference with the blank group(P>0.05).High-miao and middle-miao medicine groups'protein expression of Smad7,DCN,TGF-β1 had no obvious difference(P>0.05),but the average number of high-miao medicine groups’protein expression of Smad7,DCN was higher than the middle-miao medicine group;TGF-β1 was lower than the middle-miao medicine group.Smad7 and DCN of COPD rats models’lungs had positive correlation(P<0.01);their TGF-β1 had negative correlation(P<0.01).In the lungs of COPD rats models,content of TGF-β1 increased obviously;DCN and Smad7 decreased obviously.After treated by miao medicine,the treatment groups’TGF-β1 content de-creased obviously,DCN and Smad7 content increased obviously,and the high-miao medicine group developed especially obvious.Smad7 and DCN in COPD rats models lungs had positive correlation,but they had a negative correlation with TGF-β1.