黑咖啡提取物对人皮肤成纤维细胞胶原蛋白及衰老相关基因的影响

摘要

目的 研究黑咖啡提取物对人皮肤成纤维细胞(human skin fibroblast,HSF)胶原蛋白合成及衰老相关基因的影响.方法 体外培养HSF细胞,通过CCK-8(cell counting kit-8)法筛选黑咖啡提取物安全剂量;实验分为样品组和空白对照组,用ELISA技术检测黑咖啡提取物对细胞中Ⅰ型胶原蛋白(collagenⅠ,ColⅠ)和基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)含量的影响,用实时荧光定量聚合酶链式反应(quantitative real-time PCR,qRT-PCR)技术检测黑咖啡提取物作用后细胞中ColⅠ和MMP-1 mRNA的表达水平.结果 与空白对照组比较,黑咖啡提取物可抑制HSF细胞中MMP-1合成(P<0.05),促进ColⅠ合成(P<0.05);使ColⅠmRNA表达水平升高(P<0.01),而MMP-1 mRNA表达水平降低(P<0.01).结论 黑咖啡提取物通过促进ColⅠ和抑制MMP-1表达,对胶原蛋白代谢具有改善和调节作用,可延缓皮肤衰老.%Objective To research the effects of black coffee extract on collagen synthesis of human dermal fibroblasts (human skin fibroblast, HSF) cell and the senescence associated gene. Methods HSF cells cultured in vitro, and safe dose of black coffee extract was screened by CCK-8 (cell counting kit-8) method. The experiment was divided into experimental group and control group. The effect of black coffee extract on the content of type Ⅰcollagen (collagen Ⅰ, Col Ⅰ) and matrix metalloproteinase-1 (Matrix metalloproteinase-1, MMP-1) in HSF cells was detected by ELISA method. The expression in mRNA levels of Col Ⅰ and MMP-1 about black coffee extract was detected through qRT-PCR (quantitative real-time PCR, qRT-PCR) technology. Results Compared with the control group, black coffee extract could inhibit the synthesis of MMP-1 in HSF cells (P<0.05), promoted the synthesis of Col Ⅰ (P<0.05), increased the expression of Col Ⅰ mRNA (P<0.01), and decreased expression level of MMP-1 mRNA (P<0.01). Conclusion Black coffee extract can improve and regulate collagen metabolism by promoting Col Ⅰ and inhibiting MMP-1 expression, which has the effect of inhibiting skin aging.