尼达尼布通过TGF-β1/Smad信号通路调控人瘢痕疙瘩成纤维细胞的增殖及胶原蛋白的产生  

Nintedanib Regulates the Proliferation of Human Keloid Fibroblasts and the Production of Collagen Through the TGF-β1/Smad Signaling Pathway

作　　者：孔德祺 金文玉[1] 金哲虎[1] KONG Deqi;JIN Wenyu;JIN Zhehu(Department of Dermatology,Yanbian University Hospital,Yanji 133000,China)

机构地区：[1]延边大学附属医院皮肤科,吉林延吉133000

出　　处：《中国皮肤性病学杂志》2025年第3期258-264,共7页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金项目(82260617);吉林省教育厅科学研究项目(JJKH20240691KJ)。

摘　　要：目的探讨尼达尼布对瘢痕疙瘩成纤维细胞(keloid fibroblasts,KFs)增殖、迁移、细胞外基质(ECM)相关蛋白表达以及TGF-β1/Smad通路的影响。方法利用成品进行KFs培养。将KFs分为空白对照组、DMSO组、尼达尼布浓度组(分别为0.5、1、2、4和8μmol/L),每组培养24 h后,采用CCK-8实验检测KFs增殖情况。利用细胞划痕实验检测2μmol/L尼达尼布在不同时间段下(0、24、48 h)对KFs迁移能力的影响。根据CCK-8的结果将尼达尼布分为4组,依次为空白对照组,低、中、高浓度组(1、2、4μmol/L),通过Western blot实验分别检测CollagenⅠ、CollagenⅢ、纤连蛋白(FN)及相关蛋白α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达情况。而后再将其分为空白对照组、SB-431542(5μmol/L)干预组、尼达尼布(2μmol/L)干预组、SB-431542(5μmol/L)联合尼达尼布(2μmol/L)干预组,培养24 h。通过Western blot实验探索尼达尼布对TGF-β1/Smad信号通路蛋白的表达情况。结果根据CCK-8实验法得出尼达尼布处理KFs 24 h后,随着浓度的增加,细胞存活率呈现下降趋势,在药物浓度为8μmol/L作用24 h后,抑制KFs效果最为显著。细胞划痕实验显示,与空白对照组相比,2μmol/L尼达尼布组KFs迁移能力受到显著抑制(P<0.01);Western blot显示尼达尼布可下调CollagenⅠ、CollagenⅢ、FN、α-SMA蛋白表达,并且可降低TGF-β1/Smad通路中Smad2及Smad3磷酸化蛋白表达。结论尼达尼布可能通过TGF-β1/Smad信号通路,抑制KFs增殖、迁移及细胞外基质相关蛋白的表达。Objective To investigate the effects of nintedanib on proliferation,migration,extracellular matrix(ECM)-related protein expression,and the TGF-β1/Smad signal transduction pathway in keloid fibroblasts(KFs).Methods KFs were cultured using standard protocols.The KFs were divided into a blank control group,a DMSO group,and nintedanib concentration groups(0.5,1,2,4 and 8μmol/L).After 24 hours of culture in each group,the proliferation of KFs was assessed using the CCK-8 assay.The effect of 2μmol/L nintedanib on KFs migration at different time points(0,24,48 h)was evaluated by a cell scratch assay.Based on the CCK-8 results,nintedanib was further categorized into four groups:blank control,low,medium,and high concentrations(1,2 and 4μmol/L).Western blot analysis was performed to detect the protein expression of Collagen I,CollagenⅢ,fibronectin(FN),andα-smooth muscle actin(α-SMA).Subsequently,the cells were divided into a blank control group,an SB-431542(5μmol/L)intervention group,a nintedanib(2μmol/L)intervention group,and a combined SB-431542(5μmol/L)and nintedanib(2μmol/L)intervention group,with a further 24 hours of culture.Western blot analysis was employed to investigate the expression of proteins in the TGF-β1/Smad signaling pathway.Results According to the CCK-8 assay,the cell survival rate of KFs decreased with increasing concentrations of nintedanib after 24 hours of treatment,with the most significant inhibition observed at 8μmol/L.The cell scratch test revealed a significant inhibition of KFs migration in the 2μmol/L nintedanib group compared to the blank control group(P<0.01).Western blot analysis showed that nintedanib downregulated the protein expressions of Collagen I,CollagenⅢ,FN,andα-SMA,and also reduced the expression of phosphorylated Smad2 and Smad3 proteins in the TGF-β1/Smad pathway.Conclusion Nintedanib can inhibit the proliferation,migration,and ECM-related protein expression of KFs,potentially through mediation of the TGF-β1/Smad signaling pathway.

关 键 词：瘢痕疙瘩 尼达尼布 成纤维细胞 TGF-Β1/SMAD信号通路 细胞外基质相关蛋白 

分 类 号：R739.5[医药卫生—肿瘤]