黄芪甲苷对紫外线诱导皮肤成纤维细胞表达TGFβRⅡ与Smad7的影响(1)
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[摘要]目的：探讨黄芪甲苷对紫外线诱导皮肤成纤维细胞TGFβRⅡ、Smad7mRNA以及蛋白质表达的影响。方法：培养原代人皮肤成纤维细胞，UVA及UVB分别照射人成纤维细胞，黄芪甲苷进行干预，MTT法检测细胞增殖活性，以ELISA 法检测Smad7的生成量，半定量RT-PCR法检测TGFβRⅡ、Smad7 的mRNA表达，Western blot法检测TGFβRⅡ的蛋白表达。结果：不同浓度的黄芪甲苷干预组与正常对照组相比，成纤维细胞增殖活性随浓度增加而增强，药物浓度为40μg/ml时最显著(P<0.05)；UV辐射组与正常组相比，细胞上清液中Smad7蛋白含量明显增高，细胞内TGFβRⅡ的mRNA和蛋白表达均下降， Smad7mRNA表达增强；黄芪甲苷干预后，细胞上清液中Smad7蛋白含量下降，细胞内TGFβRⅡ的mRNA以及蛋白表达水平明显增高，Smad7mRNA表达下降(P<0.05)。结论：黄芪甲苷可能通过提高成纤维细胞的增殖活性，上调TGFβRⅡmRNA、蛋白水平以及下调Smad7 mRNA、蛋白水平的表达来对抗紫外线抑制TGF-β/Smad信号通路的传导，缓解皮肤光老化进程。
关键词：黄芪甲苷；人成纤维细胞；TGFβRⅡ；Smad7
[中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455(2011)02-0225-04
Effect of Astragaloside on TGFβRⅡ and Smad7 expression in human skin fibroblasts induced by Ultraviolet
YAN Ning1,CHEN Bin1,LI Ran1,LI Shuang-feng1,BI Zhi-gang,ZHANG Yin-di2
(1.Department of Dermatology,The First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,Jiangsu,China;
2.Institute of Clinical Pharmacology,Nanjing Medical Vniversity)
Abstract:ObjectiveTo investigate effect of Astragaloside on the expression of TGFβRⅡ,Smad7 mRNA and protein in human skin fibroblast induced by Ultraviolet.MethodsCultured human skin fibroblasts were treated with different concentrations of Astragaloside. Cellular activity was detected by MTT assay. The secretions of Smad7 in cultured fibroblasts were detected by enzvme -linked immunoadsordent assay (ELISA). The mRNA levels of Smad7 and TGFβRⅡwere determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The protein levels of TGF-βRⅡ was measured by Western blot assay.ResultsCompared with the control group, the increase of cell proliferation was observed followed the dosages-increased intervention of Astragaloside(P<0.05). The secretions and the mRNA expression of Smad7 were obviously increased, the protein expression and the mRNA levels of TGFβRⅡ was markedly decreased after UV irradiation comparing with the control group. After treated with Astragaloside, the secretions and mRNA expression of Smad7 were both decreased, the protein and the mRNA expression of TGFβRⅡwere both increased(P<0.05).ConclusionAstragaloside treatment may relieve the UV- damaged in the fibroblasts through elevating the cell proliferation activity and regulating the expression of related molecules in the TGF-β/Smad signaling pathway ......
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