贵州医科大学学报

2018, v.43;No.219(12) 1375-1379

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胸腺肽β4对TGF-β1诱导的人肺成纤维细胞的增殖、分化及促纤维化作用的影响
Effects of Thymosin beta 4 on the Proliferation,Differentiation and Fibrosis of Human Lung Fibroblasts Induced by TGF-β1

程义局;杨雪;万方;罗浩;李瑞雪;宋小霞;马文;雷显萍;黄妮雯;宋盛仁;倪木子;杜娟;张婷;
CHENG Yiju;YANG Xue;WAN Fang;LUO Hao;LI Ruixue;SONG Xiaoxia;MA Wen;LEI Xianping;HUANG Niwen;SONG Shengren;NI Muzi;DU Juan;ZHANG Ting;Department of Respiratory and Critical Diseases,Guizhou Medical University;Guizhou Medical University;Guiyang Nursing Vocational College;

摘要(Abstract):

目的:研究胸腺肽β4(Tβ4)对转化生长因子(TGF-β1)诱导的人肺成纤维细胞(HLFs)的增殖、分化、促纤维化作用的影响及机制。方法:取体外培养、对数生长期的HLFs细胞,分为对照组(等体积PBS)、TGF组(给予5μg/L TGF-β1)、Tβ4低剂量组(给予1 mg/L Tβ4及5μg/L TGF-β1)、Tβ4中剂量组(给予10 mg/L Tβ4及5μg/L TGF-β1)及Tβ4高剂量组(给予100 mg/L Tβ4及5μg/L TGF-β1),37℃培养48 h,采用Western blot检测α-SMA、Col I、p-smad2、p-smad3、smad7的蛋白表达水平,CCK-8法检测细胞增殖情况。结果:与对照组比较,TGF组HLFs细胞α-SMA、Col I、p-smad2及p-smad3蛋白表达水平升高,smad7蛋白表达水平降低(P <0. 01);与TGF组比较,Tβ4中剂量组及高剂量组HLFs细胞α-SMA、Col I、p-smad2及p-smad3蛋白表达水平降低,smad7蛋白表达水平升高(P <0. 05、0. 01); CCK-8法检测结果显示,与对照组比较,TGF组OD值显著升高(P <0. 01);与TGF组比较,Tβ4中剂量组及高剂量组HLFs细胞OD值显著降低(P <0. 01)。结论:Tβ4能抑制TGF-β1诱导的HLFs细胞增殖、分化及纤维化,其机制可能与抑制TGF-β1/smads信号通路的活化有关。
Objective: To investigate the effect of Tβ4 on the process of TGF-β1-induced proliferation,differentiation and promoting fibrosis,and its possible mechanism. Methods: HLFs lines were divided into five groups: the control group,TGF-β1-induced group,the low-dose Tβ4 group,the medium-dose Tβ4 group and the high-dose Tβ4 group. The cell viability was measured by CCK-8 assay.The α-SMA,ColI,p-smad2,p-smad3 and smad7 expressions were measured by Western blot. The expression ofα-SMA and ColI mRNA was detected by Quantitative real time PCR( qRT-PCR).Results: Compared with the control group,the expression of α-SMA,Col I,p-smad 2 and p-smad 3proteins of HLFs increased and the expression of smad7 protein decreased in TGF group. Compared with TGF group,the expressions of α-SMA,ColI,p-smad 2,p-smad3 in HLFs dercreased and the expression of smad7 increased in the medium-dose and high-dose Tβ4 groups. Compared with the controls,HLFs OD value significantly increased in TGF group,while compared with TGF group,HLFs OD value significantly decreased in the medium-dose and high-dose Tβ4 groups. Conclusion: Tβ4could inhibit the proliferation,differentiation and fibrosis of HLFs,and its mechanism may be associated with the activation inhibition of the signaling pathway of TGF-β1/smads.

关键词(KeyWords): 胸腺肽β4;人肺成纤维细胞;转化生长因子β1;信号传导;纤维化;细胞增殖
thymosin beta 4;human lung fibroblasts;transforming growth factor beta 1;signal transduction;fibrosis;cell proliferation

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基金项目(Foundation): 国家自然科学基金地区科学基金项目(81760016);; 贵州省科技计划项目[黔科合LH字(2016)7247号];; 贵州省卫计委科学技术基金项目(gzwjkj2015-1-033);; 贵州医科大学附属医院博士科研启动基金

作者(Author): 程义局;杨雪;万方;罗浩;李瑞雪;宋小霞;马文;雷显萍;黄妮雯;宋盛仁;倪木子;杜娟;张婷;
CHENG Yiju;YANG Xue;WAN Fang;LUO Hao;LI Ruixue;SONG Xiaoxia;MA Wen;LEI Xianping;HUANG Niwen;SONG Shengren;NI Muzi;DU Juan;ZHANG Ting;Department of Respiratory and Critical Diseases,Guizhou Medical University;Guizhou Medical University;Guiyang Nursing Vocational College;

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DOI: 10.19367/j.cnki.1000-2707.2018.12.003

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