积雪草苷对增生性瘢痕成纤维细胞增殖与Smad信号通路的影响

EFFECTS OF ASIATICOSIDE ON CELL PROLIFERATION AND SMAD SIGNAL PATHWAY OF HYPERTROPHIC SCAR FIBROBLASTS

目的 研究积雪草苷对增生性瘢痕成纤维细胞增殖及对 Smad信号通路的影响。 方法 采用组织块法培养人瘢痕成纤维细胞 ,将成纤维细胞分为用与不用积雪草苷两组 (实验组和对照组 ) ,4 8小时后采用逆转录 -聚合酶链反应 ( reverse transcription- polymerase chain reaction,RT- PCR)观察 Smad2及 Smad7m RNA表达量的改变。应用流式细胞仪分析、免疫细胞化学及蛋白印迹 ( Western blot)技术结合光密度扫描分析 ,观察积雪草苷对瘢痕成纤维细胞周期及磷酸化 Smad2和 Smad7的改变。 结果 实验组可见瘢痕中成纤维细胞从 S期进入 M期细胞减少 ,受到抑制 ;成纤维细胞中 Smad2的含量及 Smad2 m RNA的表达实验组较对照组改变不明显 ,但 Smad7的含量及 Smad7m RNA的表达实验组与对照组分别为 ( 1.33± 1.2 6 ) %、( 5 0 .80± 2 2 .4 0 ) %及 ( 9.15± 3.36 ) %、( 32 .18± 17.84 ) % ,二者两组差异均有统计学意义 ( P<0 .0 5 )。 结论 积雪草苷抑制瘢痕的作用是通过 Smad通路 。

Objective To investigate the effects of asiaticoside on the proliferation and the Smad signal pathway of the hypertrophic scar fibroblasts. Methods The hypertrophic scar fibroblasts were cultured with tissue culture method. The expressions of Smad2 and Smad7 mRNA after asiaticoside treatment were determined by reverse transcription polymerase chain reaction 48 hours later. The cell cycle, the cell proliferation, the cell apoptosis and the expression of phosphorylated Smad2 and Smad7 with(experimental group) or without(control group) asiaticoside were detected with flow cytometry, immunocytochemistry and Western blot. Results Asiaticoside inhibited the hypertrophic scar fibroblasts from phase S to phase M. The Smad7 content and the expression of Smad7 mRNA were (1 33±1 26)% and (50 80±22 40)% in experimental group, and (9 15±3 36)% and (32 18±17 84)% in control group; there were significant differences between two groups ( P<0 05) . While the content and the mRNA expression of Smad2 had no significant difference between two groups. Conclusion Asiaticoside inhibits the scar formation through Smad signal pathway.