| 引用本文: | 侯欢,刘金成,陈蓉.蛇床子素通过抑制小鼠心肌成纤维细胞TGF-β1/Smad信号通路降低胶原表达的研究[J].中国现代应用药学,2022,39(19):2437-2443. |
| HOU Huan,LIU Jincheng,CHEN Rong.Study on Osthole Decreased Collagen Expression by Inhibiting TGF-β1/Smad Signaling Pathway in Mouse Cardiac Fibroblasts[J].Chin J Mod Appl Pharm(中国现代应用药学),2022,39(19):2437-2443. |
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| 摘要: |
| 目的 研究蛇床子素降低真核载体pcDNA3.1(+)-TGF-β1转染的小鼠心肌成纤维细胞(cardiac fibroblasts，CFs)胶原的表达是否与抑制TGF-β1/Smad信号通路相关。方法 用真核表达载体pcDNA3.1(+)-TGF-β1转染小鼠CFs，然后在TGF-β1受体抑制剂SB431542预处理或不预处理的情况下，用不同浓度的蛇床子素处理CFs 24 h。ELISA检测上清TGF-β1水平，Western blotting检测细胞内α-SMA、collagen Ⅰ、collagen Ⅲ、Smad2/3、p-Smad2/3、Smad4和Smad7的蛋白表达。结果 真核表达载体pcDNA3.1(+)-TGF-β1转染CFs后，TGF-β1释放量增加，提示建立了TGF-β1过表达的细胞模型。TGF-β1过表达的CFs经蛇床子素(5~20 μmol·L-1)处理24 h后，TGF-β1、α-SMA、collagenⅠ、collagenⅢ、p-Smad2/3和Smad4蛋白表达降低，Smad7蛋白表达上升。TGF-β1受体抑制剂SB431542预处理2 h后，蛇床子素对α-SMA、collagen Ⅰ、collagen Ⅲ、Smad2/3、p-Smad2/3和Smad4蛋白表达的抑制作用进一步增强。结论 蛇床子素可以通过直接抑制TGF-β1的产生和Smad通路下调α-SMA和胶原蛋白的表达，这可能是其抗纤维化的机制之一。 |
| 关键词: 蛇床子素 转化生长因子-β1/Smad信号通路 α-平滑肌肌动蛋白 胶原蛋白Ⅰ 胶原蛋白Ⅲ |
| DOI：10.13748/j.cnki.issn1007-7693.2022.19.002 |
| 分类号:R285.5 |
| 基金项目:国家自然科学基金项目(81302772);江苏省卫生计生委青年医学重点人才培养项目(QNRC2016716);苏州市科技发展计划项目(SYSD2020101) |
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| Study on Osthole Decreased Collagen Expression by Inhibiting TGF-β1/Smad Signaling Pathway in Mouse Cardiac Fibroblasts |
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HOU Huan1,2, LIU Jincheng3, CHEN Rong1
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1.Department of Pharmacology, The First Affiliated Hospital of Soochow University, Suzhou 215006, China;2.Department of Pharmacology, College of Pharmaceutical Science, Soochow University, Suzhou 215123, China;3.Department of Pharmacology and Traditional Chinese Medicine, School of Chinese Materia Medica, China Pharmaceutical University, Nanjing 211198, China
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| Abstract: |
| OBJECTIVE To explore whether the reduction of collagen expression in mouse cardiac fibroblasts (CFs) transfected with eukaryotic vector pcDNA3.1(+)-TGF-β1 directly associated with inhibition of TGF-β1/Smad signaling pathway.METHODS The mouse CFs were transfected with eukaryotic vector pcDNA3.1(+)-TGF-β1,and the TGF-β1-transfected CFs were then treated with different concentrations of osthole for 24 h in the presence or absence of pretreatment with TGF-β1 receptor Ⅰ inhibitor SB431542.The supernatant TGF-β1 level was determined by ELISA method,and the intracellular α-SMA,collagen Ⅰ,collagen Ⅲ,Smad2/3,p-Smad2/3 and Smad4 and Smad7 protein expressions were determined by Western blotting.RESULTS After transfection of CFs with eukaryotic vector pcDNA3.1(+)-TGF-β1,the release of TGF-β1 was increased,indicating that the cell model of TGF-β1-overexpression was established.Following treatment of TGF-β1-overexpressed CFs with osthole 5-20 μmol·L-1 for 24 h,the TGF-β1,α-SMA,collagen Ⅰ,collagen Ⅲ,p-Smad2/3 and Smad4 protein expressions were decreased,while Smad7 protein expressions were increased.After pretreatment with TGF-β1 receptor Ⅰ inhibitor SB431542 for 2 h,the inhibitory effects of osthole on α-SMA,collagen Ⅰ,collagen Ⅲ,Smad2/3,p-Smad2/3 and Smad4 protein expressions were further enhanced.CONCLUSION Osthole can downregulate the expressions of α-SMA and collagen proteins by directly inhibiting TGF-β1 production and the Smad pathway,which might be one of its anti-fibrotic mechanisms. |
| Key words: osthole TGF-β1/Smad pathway α-smooth muscle actin (α-SMA) collagen Ⅰ collagen Ⅲ |
