摘要
目的研究丹参酮ⅡA(TSA)对TGF-β1诱导的人皮肤成纤维细胞(HSF)的影响及作用机制。方法采用CCK-8法测定不同浓度TSA(2.5、5、10、20μmol/L)干预TGF-β1诱导HSF细胞增殖的影响;采用平板克隆形成实验分别分析不同浓度TSA处理48 h后HSF细胞的克隆形成能力的变化;Western blotting检测HSF细胞中TGF-β1/Smad通路蛋白及α-SMA、VEGFA和COLI蛋白的表达水平。结果CCK-8和平板克隆形成实验结果表明,TSA显著抑制了HSF细胞的增殖和克隆形成能力(P<0.01);Western blotting结果显示,TSA各浓度组能够显著抑制p-Smad2和p-Smad3蛋白水平,下调p-Smad2/Smad2的比值(P<0.01)。5、10、20μmol/L TSA组明显下调p-Smad3/Smad3的比值;另外,HSF细胞中α-SMA、VEGFA和COLI蛋白表达水平随着TSA浓度的升高而显著降低(P<0.01)。结论TSA具有抑制HSF细胞增殖的能力,其作用机制可能与TGF-β1/Smad通路有关。
Objective To study the effect and mechanism of tanshinone ⅡA on human skin fibroblasts cell(HSF). Methods CCK-8 method was used to determine the effect of different concentrations of TSA on the proliferation of HSF induced by TGF-β1. The plate cloning ability of HSF treated with different concentrations of TSA(2.5, 5, 10 and 20 μmol/L) for 48 h were analyzed by plate clonogenesis assay. The protein expression of TGF-β1/Smad signaling pathway proteins and α-SMA, VEGFA and COL I were further measured by Western blotting. Results CCK-8 and plate clonognesis assay results showed that TSA significantly inhibited the proliferation and colony forming efficiency of HSF(P < 0.01). Western blotting results revealed that each concentration group of TSA significantly inhibited the protein levels of p-Smad2 and p-Smad3, and down-regulated the ratio of p-Smad2/Smad2(P < 0.01). The ratio of p-Smad3/Smad3 was significantly decreased in 5, 10 and 20 μmol/L TSA groups. Additionally, the expression levels of α-SMA, VEGFA and COL I in HSF decreased significantly with the increase of TSA concentration(P < 0.01). Conclusion TSA exhibits the inhibitory effect on proliferation of HSF, and its mechanism may be related to TGF-β1/Smads signaling pathway.
作者
许小琪
韩兵
赖建辉
林宇建
时军
XU Xiao-qi;HAN Bing;LAI Jian-hui;LIN Yu-jian;SHI Jun(Department of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;Plastic and Reconstructive Surgery,First Affiliated Hospital of SUN Yat-sen University,Guangzhou 510080,China;Guangdong Engineering&Technology Research Center of Topical Precise Drug Delivery System,Guangzhou 510006,China)
出处
《中草药》
CAS
CSCD
北大核心
2020年第18期4685-4690,共6页
Chinese Traditional and Herbal Drugs
基金
广东省自然科学基金项目(2018A0303130234)
广东省高等学校优秀青年教师培养计划项目(YQ2015099)。
