芒柄花黄素对人增生性瘢痕成纤维细胞生长的作用

Effect of formononetin on growth of human hypertrophic scar fibroblasts

目的探讨芒柄花黄素对人增生性瘢痕成纤维细胞生长的作用。方法获取手术切除的人增生性瘢痕组织以及局部皮瓣转移修复后剩余的正常皮肤,提取成纤维细胞。通过MTT试验检测不同浓度的芒柄花黄素对成纤维细胞增殖的作用,应用流式细胞术对芒柄花黄素处理后的成纤维细胞周期进行分析。通过Annexin V-FITC/PI双染色法和Hoechest 33258染色观察成纤维细胞的凋亡情况。通过Western blot与RT-PCR试验,检测芒柄花黄素对细胞周期蛋白Cyclin D1和凋亡调节蛋白Bcl-2的作用。结果与对照组比较,芒柄花黄素对正常皮肤成纤维细胞的增殖影响较小,对增生性瘢痕成纤维细胞的增殖具有明显的抑制作用,且药物浓度越高、作用时间越长,抑制作用越强。成纤维细胞大部分阻滞于G_1期,S期的细胞减少,细胞周期的进程受到阻断。芒柄花黄素能够通过抑制细胞周期蛋白Cyclin D1来影响细胞周期的进程,将细胞周期阻滞于G_1期;通过抑制抗凋亡蛋白Bcl-2的表达诱导成纤维细胞的凋亡,从而实现对细胞生长的抑制作用。结论芒柄花黄素能够抑制人增生性瘢痕成纤维细胞的增殖并诱导其凋亡,可考虑用于增生性瘢痕的预防与临床治疗。

Objective To explore the effect of formononetin on growth of human hypertrophic scar fibroblasts.Methods Human hypertrophic scar tissue and the remaining normal skin after local skin flap transfer and repair were obtained and fibroblasts were extracted.Effect of different concentrations of formononetin on fibroblasts proliferation was tested by MTT assay.The fibroblasts cycle after formononetin treatment was analyzed by flow cytometry.The apoptosis of fibroblasts was observed by Annexin V-FITC/PI and Hoechest 33258 staining.Effect of formononetin on Cyclin D1 and Bcl-2 was detected by Western blot and RT-PCR.Results Compared with control group,formononetin had little effect on the proliferation of fibroblasts from normal skin,but it inhibited the proliferation of the hypertrophic scar fibroblasts significantly.The higher the drug concentration,the longer the action time,the stronger the inhibitory effect.A majority of the fibroblasts were blocked in G1 phase and reduced in S phase.The process of cell cycle was blocked.Formononetin affected the cell cycle process through inhibiting the expression of Cyclin D1,and the cell cycle was blocked at G1 phase.Formononetin induced the apoptosis of fibroblasts by inhibiting the expression of anti-apoptotic protein of Bcl-2,so that the inhibition of cell growth could be realized.Conclusion Formononetin can inhibit the proliferation and induce the apoptosis of fibroblasts,which can be considered for prevention and clinical treatment of hypertrophic scar.