【摘要】 目的 基于TGF-β1/Smads信号通路探讨毛蕊花糖苷对人增生性瘢痕成纤维细胞的作用及其机制研究。方法 CCK8法检测毛蕊花糖苷对增生性瘢痕成纤维细胞的半数抑制浓度。将实验分为对照组、毛蕊花糖苷低浓度组（40μg/ml毛蕊花糖苷）、毛蕊花糖苷组高浓度（80μg/ml毛蕊花糖苷）。划痕实验检测不同浓度毛蕊花糖苷对增生性瘢痕成纤维细胞的迁移能力的影响，流式细胞术检测不同浓度毛蕊花糖苷对增生性瘢痕成纤维细胞的细胞周期与凋亡的影响。选用最佳浓度的TGF-β1作为模型组，再将实验分为对照组、模型组：5 ng/ml TGF-β1；低浓度实验组（TGF-β1+40μg/ml毛蕊花糖苷）；高浓度实验组（TGF-β1+80μg/ml毛蕊花糖苷）。实时荧光定量PCR、Western blot法检测四组Smad2、Smad3和COL-1的m RNA及蛋白表达变化。结果 毛蕊花糖苷对增生性瘢痕成纤维细胞的半数抑制浓度为79.54μg/ml。毛蕊花糖苷低、高浓度组增生性瘢痕成纤维细胞迁移距离小于对照组（P<0.05）。与对照组比较，毛蕊花糖苷低、高浓度组均能将增生性瘢痕成纤维细胞的细胞增殖周期阻滞在G0/G1期（P<0.05）。与对照组比较，毛蕊花糖苷高浓度组成纤维细胞凋亡率升高（P<0.01）。与对照组比较，模型组中Smad2、Smad3及COL-1 m RNA的表达量升高（P<0.05）；与模型组比较，低、高浓度实验组Smad2、Smad3及COL-1 m RNA表达量降低（P<0.05）。与对照组比较，模型组P-Smad2/3、COL-1蛋白表达水平升高（P<0.05）；与模型组比较，高浓度实验组P-Smad2/3表达水平降低，低、高浓度实验组COL-1蛋白表达水平降低（P<0.05）。结论 毛蕊花糖苷可通过抑制TGF-β1/Smads信号通路进而抑制增生性瘢痕成纤维细胞的活化和增殖。更多还原

【Abstract】 Objective To explore the role of verbascoside on human hypertrophic scar fibroblasts and its mechanism based on TGF-β1/Smads signaling pathway. Methods The half-inhibitory concentration of verlein on hypertrophic scar fibroblasts was determined by CCK8 method. The experiment was divided into control group and low concentration group(40 μg/ml uerbascoside), high concentration of verminin group(80 μg/ml verbascoside). The effects of different concentrations of uerbascoside on the migration ability of fibroblasts in hypertrophic scar were detected by scratch test, and the changes of cell cycle and apoptosis of fibroblasts in hypertrophic scar were detected by flow cytometry. The optimal concentration of TGF-β1 was selected as the model group, and then the experiment was divided into control group and model group: 5 ng/ml TGF-β1; low concentration experimental group(TGF-β1+40 μg/ml verbascoside); high concentration experimental group(TGF-β1+80 μg/ml verbascoside). The m RNA and protein expression of Smad2, Smad3, and COL-1 in the four groups were detected by real-time fluorescence quantitative PCR and Western blot. Results The median inhibitory concentration of verlein on hypertrophic scar fibroblasts was 79.54 μg/ml. The migration distance of hypertrophic scar fibroblasts in low and high concentration of verbascoside was smaller than that in control group(P<0.05). Compared with the control group, the cell proliferation cycle of hypertrophic scar fibroblasts could be blocked in G0/G1phase in both low and high concentration of vervascoside(P<0.05). Compared with the control group, the apoptosis rate of fiber cells with high concentration of verbascoside was increased(P<0.01).Compared with control group, m RNA expressions of Smad2, Smad3, and COL-1 in model group were increased(P<0.05).Compared with model group, m RNA expressions of Smad2, Smad3, and COL-1 in low and high concentration groups were decreased(P<0.05). Compared with control group, the expression levels of P-SMAD2/3 and COL-1 protein in model group were increased(P <0.05). Compared with model group, the expression level of P-SMAD2/3 in high-concentration experimental group was decreased, and the COL-1 expression level in low concentration and high-concentration experimental group was decreased(P<0.05). Conclusion Verbascoside can inhibit hypertrophic scar fibroblasts activation and proliferation by blocking TGF-β1/Smad signaling pathway.更多还原