5,7,4′-三羟基异黄酮对增生性瘢痕成纤维细胞增殖细胞核抗原表达及细胞周期的影响

目的 观察5,7,4′-三羟基异黄酮(genistein)对增生性瘢痕成纤维细胞增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)及细胞周期的影响,探讨5,7,4′-三羟基异黄酮抑制增生性瘢痕成纤维细胞增殖的机制.方法 分离培养人增生性瘢痕成纤维细胞,分别加入25、50、100 μmol/L浓度的5,7,4′-三羟基异黄酮共培养48 h,免疫细胞化学法观察成纤维细胞PCNA蛋白的表达,流式细胞术检测细胞周期的变化.结果 各组5,7,4′-三羟基异黄酮作用后细胞PCNA的表达均降低(P＜0.05),50 μmol/L及100 μmol/L浓度组的抑制作用最为显著(P＜0.01);随药物浓度的增加,G0～G1期细胞比例逐渐下降,G2～M期细胞比例增加,表明细胞分裂受到抑制;100 μmol/L组的S期细胞数量比例也有增加,并于G1期前出现亚二倍体凋亡峰.结论 5,7,4′-三羟基异黄酮可通过影响细胞分裂与DNA合成抑制瘢痕增生.

Effects of genistein on PCNA expression and cell cycle in human hypertrophic scar fibroblasts in vitro

Objective To observe the effects of genistein on PCNA expression and cell cycle in fibroblasts derived from human hypertrophic scar in order to explore the mechanism of its inhibition on hypertrophic scar (HS) fibroblast proliferation. Methods The human hypertrophic scar fibroblasts were cultured in vitro. Genistein with various concentrations (25, 50, 100 μmol/L) was co-cultured in the medium for 48 hours. The expression of PCNA was detected with immunocytochemical staining method and the cell cycle was measured with flow cytometry. Results Genistein could significantly decrease PCNA expression in HS fibroblasts, especially when its concentration at 50 μmol/L or 100 μmol/L. The cell percentage of G0～G1 phase decreased with drug′s concentration, and G2～M percentage increased conversely, implying the suspension of mitosis. In 100 μmol/L group, most cells blocked at S phase and a hypodiploid apoptosis peak could be observed ahead of G1 phase. Conclusion Genistein can inhibit the proliferation of human hypertrophic scar by blocking cell division as well as decreasing DNA synthesis.