姜黄素对人增生性瘢痕成纤维细胞生长及功能的影响

作者	单位
康建毅	重庆大学生物工程学院
黄宏	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室
朱方强	第三军医大学大坪医院泌尿外科
张波	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室
王建民	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室
蒋建新	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室
付小兵	解放军总医院304临床部全军创伤修复重点实验室
朱佩芳	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室
王正国	第三军医大学大坪医院野战外科研究所创伤、烧伤与复合伤国家重点实验室

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中文摘要:

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。

英文摘要:

Objective To seek an effective drug for treatment of human hyperplastic scar through studying the effects of curcumin on fibroblast growth and collagen synthesis.Methods Fibroblasts derived from scar tissue and from normal epidermal tissue were isolated and cultured separately with tissue-block method,their morphology were observed under invert phase contrast microscope,their growth curve was drawn respectively to determine the speed of growth.Then,fibroblasts from scar were stimulated with curcumin in different concentrations(0,12.5,25,50 and 100 μmol/L) for detecting the inhibitory effect of curcumin on growth of fibroblasts using MTT methods and that on activity of precollagen α-1 gene transcription in fibroblast was detected by RT-PCR.Results The cell growth curve showed that double-multiplying time was 5 days in fibroblasts from scar and 4 days in those from normal dermis,showing significant difference between them(P<0.05).MTT showed that curcumin in 12.5 μmol/L showed a cell proliferation enhancing trend,and its absorbance value was significantly higher than that in the normal group,but the effect turned to inhibition when concentration increased to over 25-100 μmol/L,and became significant inhibition at concentration of 50 and 100 μmol/L.Besides,curcumin also showed markedly inhibition on collagen type Ⅰ synthesis in fibroblasts(P<0.01).Conclusion High concentration curcumin can inhibit effectively the fibroblast proliferation and collagen Ⅰ synthesis in hyperplastic scar,therefore,may has therapeutic effect on the disease in human being.

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