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

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 Tβ4on 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 Tβ4group,the medium-dose Tβ4group and the high-dose Tβ4group.The cell viability was measured by CCK-8assay.Theα-SMA,ColI,p-smad2,p-smad3and smad7expressions were measured by Western blot.The expression ofα-SMA and ColI m R N A was detected by Quantitative real time P C R(q R T-P C R).Results:Compared with the control group,the expression ofα-SMA,ColI,p-smad2and proteins of HLFs increased and the expression of smad7protein decreased in TGF group.Compared with TGF group,the expressions ofα-SMA,Coll,p-smad2,p-smad3in HLFs dercreased and the expression of smad7increased in the medium-dose and high-dose Tβ4groups.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 Tβ4groups.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.