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目的探讨槲皮素对人体瘢痕疙瘩成纤维细胞在生长增殖、细胞凋亡方面的作用,为临床瘢痕疙瘩的治疗提供实验基础。方法MTT法测定瘢痕疙瘩成纤维细胞干预前后的生长曲线;光学显微镜观察干预后细胞的形态学变化;流式细胞仪分析成纤维细胞在干预前后的细胞周期变化;原位细胞凋亡检测成纤维细胞干预后的细胞凋亡率;免疫细胞化学和蛋白印迹实验检测成纤维细胞干预前后p53、bcl-2、bax蛋白表达水平的变化。结果MTT法显示槲皮素对体外培养的瘢痕疙瘩成纤维细胞有抑制作用,抑制效应呈剂量时间依赖关系;槲皮素作用后48h瘢痕疙瘩成纤维细胞发生G2/M期阻滞,并且出现细胞凋亡,光镜可见细胞形态学上出现细胞皱缩,染色质浓集;流式细胞仪结果显示有凋亡峰出现;原位细胞凋亡检测显示凋亡率为16.5%±2.7%。槲皮素作用后细胞p53蛋白和bax蛋白表达升高,bcl-2蛋白表达降低。结论槲皮素通过抑制细胞增殖和促进细胞凋亡来抑制瘢痕疙瘩成纤维细胞的生长。
Objective To investigate the effect of quercetin on growth and proliferation of human keloid fibroblasts and to provide experimental basis for the treatment of clinical keloids. Methods MTT assay was used to determine the growth curve of keloid fibroblasts before and after intervention; light microscopy was used to observe the morphological changes of the cells after intervention; flow cytometry was used to analyze the cell cycle changes of fibroblasts before and after intervention; in situ cell apoptosis was detected by fibroblasts The cell apoptosis rate after cell intervention; Immunocytochemistry and Western blotting experiments were used to detect the changes of p53, bcl-2 and bax protein expression levels after fibroblasts intervention. Results MTT assay showed that quercetin inhibited keloid fibroblasts cultured in vitro and the inhibitory effect was dose-dependent; quercetin induced G2/M arrest and cells appeared 48 hours after quercetin treatment. Apoptosis, light microscopy showed cell shrinkage and chromatin condensation on cell morphology; flow cytometry showed apoptotic peaks; in situ apoptosis showed apoptotic rate of 16.5%±2.7%. After treatment with quercetin, the expression of p53 protein and bax protein increased, and the expression of bcl-2 protein decreased. Conclusion Quercetin inhibits the growth of keloid fibroblasts by inhibiting cell proliferation and promoting apoptosis.