松脂醇二葡萄糖苷对光老化HDF细胞ER/TGF-β1/Smads信号通路的调控机制

Regulatory role of pinoresinol diglucoside in the ER/TGF-β1/Smads signaling pathways in photo-aged HDF cells

目的从分子生物学水平探讨杜仲中松脂醇二葡萄糖苷(PDG)对光老化人真皮成纤维细胞(HDF)ER/TGF-β1/Smads信号通路的调控机制。方法建立以长波紫外线模型损伤HDF细胞。给予不同浓度PDG和通路阻断剂进行干预,用MTT检测细胞增殖率,Real time-PCR和Western blot法检测各组细胞内含有的信号转化生长因子(TGF-β1)、转导分子(Smad3)、I型胶原(COL1A1)对应mRNA和蛋白的表达。结果与空白组比较,模型组增殖率及TGF-β1、Smad3、COL1A1 mRNA和蛋白表达均显著降低(P <0.01);与模型组比较,各给药组增殖率和TGF-β1、Smad3、COL1A1mRNA和蛋白表达均显著升高(P<0.01);与PDG高浓度组比较,TGF-β1、Smad3、ER三个阻断剂组相对应的TGF-β1、Smad3和COL1A1相应mRNA和蛋白表达均显著降低(P <0.01)。结论 PDG能增加HDF细胞TGF-β1、Smad3、COL1A1对应mRNA和蛋白表达,对光老化HDF细胞胶原合成有促进作用;但这种作用能被TGF-β1、Smad3和ER阻断剂所抑制, PDG促进胶原合成的作用机制可能与ER/TGF-β1/Smads信号通路有关。

Objective To study the effect of pinoresinol diglucoside （PDG） on the ER/TGF-β1/Smads signaling pafflways in photo-aged human dermal fibroblasts （HDF） cells. Methods A long wave ultraviolet （UVA） model was established to damage file HDF cells, and different concentrations of PDG and ER/TGF-β1/Smads signaling pathway inhibitors were added. The cell proliferation rate was detected by MTT, and the expression of signal transforming growth factor （TGF- β1）, transduced molecule （Smad3） and type I collagen （COL 1A 1） expression were detected by RT-PCR and Western blot method. Results Compared wiffl file control group, file proliferation rate and file expression of TGF-β1, Smad3, COL1A1 mRNA and protein in file UVA irradiated model group were significantly decreased （P 〈 0.01）. Compared wiffl file model group, the proliferation rate and TGF-β1, Smad3, and COL1A1 expression were significantly increased after PDG treatment （P 〈 0.01）. Compared with file high concentration of PDG group, the expression of TGF-β1, Smad3, and COL1A1 expression were significantly decreased, respectively, upon file treatment with respective inhibitor （P 〈 0.01）. Conclusion PDG increased file expression of TGF- β1, Smad3 and COL1A1 in HDF cells after UVA irradiation, and this effect can be inhibited by TGF-β1, Smad3 and ER inhibitor.