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目的探讨姜黄素对人增生性瘢痕(hyperplastic scar)成纤维细胞(fibroblast,FB)增殖及胶原合成的影响,以寻找治疗人增生性瘢痕的有效药物。方法应用组织块法分离培养瘢痕和正常真皮组织的FB,在倒置相差显微镜下观察细胞形态,应用细胞计数方法绘制细胞生长曲线,测定细胞生长速度;采用不同浓度的姜黄素(0、12.5、25、50和100μmol/L)刺激瘢痕FB,采用四氮甲基唑盐比色法(MTT法)检测姜黄素对FB生长的抑制效应,采用RT-PCR检测姜黄素对FBα1(Ⅰ)前胶原基因的转录活性的影响。结果细胞生长曲线显示:增生性瘢痕FB倍增时间约为5天,正常皮肤FB后细胞倍增时间为4天,两者比较,差异有统计学意义(P<0.05);MTT检测结果显示:12.5μmol/L姜黄素作用于增生性瘢痕FB,表现为增殖倾向,其光吸收值(A值)显著地高于正常对照组。而在25~100μmol/L的浓度范围内,随着姜黄素药物浓度的增加,其抑制作用逐渐增强,50μmol/L和100μmol/L姜黄素能显著抑制瘢痕FB增殖(P<0.05);此外,50μmol/L和100μmol/L姜黄素能够显著抑制瘢痕FBⅠ型胶原合成(P<0.01)。结论较高浓度的姜黄素能有效抑制增生性瘢痕FB增殖和Ⅰ型胶原合成,对增生性瘢痕可能具有治疗作用。
Objective To investigate the effects of curcumin on the proliferation and collagen synthesis of hyperplastic scar fibroblasts (FB) in order to find effective drugs for the treatment of hypertrophic scars in humans. Methods The FB of scars and normal dermis were isolated and cultured by tissue block method. The morphology of the cells was observed under an inverted phase contrast microscope. The cell growth curve was drawn using a cell counting method to determine the cell growth rate; different concentrations of curcumin (0, 12.5, 25) were used. 50, 100 and 100 μmol/L) were used to stimulate scar FB. The inhibitory effect of curcumin on FB growth was detected by tetrazolium methyl ester colorimetric method (MTT method). RT-PCR was used to detect curcumin on FBα1(I) procollagen gene. The effect of transcriptional activity. Results The cell growth curve showed that the FB doubling time of hypertrophic scar was about 5 days, and the doubling time of FB after normal skin was 4 days. There was a statistically significant difference between the two groups (P<0.05). The results of MTT assay showed that: 12.5 μmol /L curcumin acted on the hypertrophic scar FB, showing proliferative tendency, and its light absorption value (A value) was significantly higher than that of the normal control group. In the concentration range of 25~100μmol/L, with the increase of curcumin concentration, its inhibitory effect was gradually enhanced, 50μmol/L and 100μmol/L curcumin could significantly inhibit the proliferation of scar FB (P<0.05); 50μmol/L and 100μmol/L curcumin could significantly inhibit the synthesis of scar FBI collagen (P<0.01). Conclusion Higher concentrations of curcumin can effectively inhibit the proliferation of FB and the synthesis of type I collagen in hypertrophic scars, and may have a therapeutic effect on hypertrophic scars.