目的 观察豆蔻明(Cardamonin,CDN)对肺纤维化小鼠的作用,并探究其对转化生长因子-β1(transforming growth factor-β1,TGF-β1)/smad信号通路的影响.方法 将小鼠随机分为生理盐水(Sham)组、博莱霉素(Bleomycin,BLM)组、豆蔻明低剂量(low dose of Cardamonin,CDN-L)组、豆蔻明中剂量(medium dose of Cardamonin,CDN-M)组、豆蔻明高剂量(high dose of Cardamonin,CDN-H)组、地塞米松(dexamethasone,DXM)组;单次气管注射BLM诱导肺纤维化,计算肺指数;酶联免疫吸附法测血清肿瘤坏死因子(tumor necrosis factor-α,TNF-α)水平;试剂盒测肺组织羟脯氨酸(hydroxyproline,HYP)含量,苏木精-伊红、马松染色观察肺炎症程度及纤维化情况,RT-PCR 法检测肺组织TGF-β1/smad信号通路相关基因表达情况.结果 与Sham组相比,BLM组小鼠肺指数、Szapiel评分、Ashcroft 评分明显升高(P<0.05),肺部炎症程度和纤维化程度严重.血清中TNF-α含量升高(P<0.05)、肺组织HYP水平升高(P<0.05),TGF-β1[(1.02±0.21)vs.(3.25±0.14)]、smad2[(1.00±0.05)vs.(1.63±0.09)]、smad3[(1.00±0.06)vs.(1.82±0.07)]、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)[(1.00±0.10)vs.(2.15±0.10)]mRNA表达升高(P<0.05),上皮-钙黏蛋白(E-Cadherin)[(1.01±0.16)vs.(0.57±0.09)]mRNA水平降低(P<0.05).CDN-M、CDN-H干预可减低小鼠肺指数,减轻炎性和肺纤维化,降低TNF-α、HYP水平(P<0.05),同时下调肺纤维小鼠肺组织中TGF-β1、smad2、smad3、α-SMA 表达,上调E-Cadherin表达.CDN-L无明显治疗效果,CDN-H治疗作用与DXM相似.结论 CDN可能通过TGF-β1/Smad通路介导上皮-间充质转化而发挥抗纤维化作用.
Therapeutic effect and mechanism of Cardamonin on pulmonary fibrosis
Objective To observe the effect of Cardamonin(CDN)on pulmonary fibrosis in mice,and explore the effect of CDN on transforming growth factor-β1(TGF-β1)/Smad signaling pathway.Methods The mice were grouped into:Sham group,Bleomycin(BLM)group,low dose of Cardamonin(CDN-L)group,medium dose of Cardamonin(CDN-M)group,high dose of Cardamonin(CDN-H)group and Dexamethasone(DXM)group.Injectioning of BLM induce pulmonary fibrosis in mice,the lung index was measured.Enzyme-linked immunosorbent assay kit measured serum tumor necrosis factor-α(TNF-α)levels,the hydroxyproline(HYP)content in lung tissue was detectioned by kits.Pathological changes were observed by Htoxylin Eosin and Masson staining,and the level of genes related to TGF-β1/smad signaling pathway was detected by RT-PCR.Results Compared with the Sham group,the lung index,Szapiel score and Ashcroft score of the BLM group were significantly increased(P<0.05),and the degree of pulmonary inflammation and fibrosis was more severe.The levels of TNF-α in serum and HYP in lung tissue were increased(P<0.05),lung tissue TGF-β1[(1.02±0.21)vs.(3.25±0.14)],smad2[(1.00±0.05)vs.(1.59±0.20)],smad3[(1.00±0.06)vs.(1.59±0.20)],α-smooth muscle actin(α-SMA)[(1.00±0.10)vs.(2.15±0.10)and E-Cadherin[(1.01±0.16)vs.(0.57±0.09)]mRNA level decreased(P<0.05).The intervention of CDN-M and CDN-H could decrease the lung index,alleviate the inflammation and Pulmonary fibrosis,and decrease the levels of TNF-α and HYP(P<0.05).The expression of TGF-β1,smad2,smad3 and α-SMA in lung tissue of mice with pulmonary fibrosis was down-regulate.CDN-L had no significant effect,while CDN-H had similar effects to DXM.Conclusion Cardamonin may play an anti-fibrotic role by mediating epithelial-mesenchymal transition through the TGF-β1/Smad pathway.
CardamoninPulmonary fibrosisTGF-β1/SmadEpithelial-mesenchy mal transition
万方数据
维普
