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
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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.
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.
引文
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[3]新华网.中国学者揭示咖啡因减肥作用新机制[J].饮料工业,2017,20(4):19.Xinhua Net.Chinese scholars reveal new mechanisms for caffeine weight loss[J].Beverage Ind,2017,20(4):19.
[4]Nkondjock A.Coffee consumption and the risk of cancer:An overview[J].Cancer Letter,2009,277(2):121-125.
[5]杨剀舟,翟晓娜,杜秉健,等.咖啡中功能性成分分离检测技术及安全性评价[J].食品科学,2014,35(3):243-252.Yang KZ,Zhai XN,Du BJ,et al.Recent advances in technologies for the separation,determination and safety evaluation of functional components in coffee[J].Food Sci,2014,35(3):243-252.
[6]范金波,蔡茜彤,冯叙桥,等.咖啡酸体外抗氧化活性的研究[J].中国食品学报,2015,3(15):65-73.Fan JB,Cai XT,Feng XQ,et al.Study on antioxidant activity of caffeic acid in vitro[J].Chin J Food Sci,2015,3(15):65-73.
[7]林淑媛,王联辉,林苑晖,等.不同制备法制得咖啡之抗氧化性及咖啡因含量[J].台湾农业化学与食品科学,2009,47(5):268-275.Lin SY,Wang LH,Lin YH,et al.Antioxidative activity and caffeine content of coffee from different preparation methods[J].Taiwan Agric Chem Food Sci,2009,47(5):268-275.
[8]丁莉,周向阳.中国咖啡产业竞争力分析及展望[J].农业展望,2017,(10):52-57.Ding L,Zhou XY.Analysis and prospect of coffee industry competitiveness in China[J].Agric Outlook,2017,(10):52-57.
[9]文志华,毕晓菲,鲁维艳.中国咖啡消费趋势浅谈[J].农产品加工,2018,1(448):69-70.Wen ZH,Bi XF,Lu WY.Discussion on the trend of Chinese coffee consumption[J].Farm Prod Process,2018,1(448):69-70.
[10]王曦,李利.成纤维细胞与皮肤老化[J].中国美容医学,2005,14(2):243-245.Wang X,Li L.Fibroblast and aging of skin[J].Chinese J Aesthet Med,2005,14(2):243-245.
[11]Chung JH,Seo JY,Choi HR,et al.Modulation of skin collagen metabolism in aged and photoaged human skin in vivo[J].J invest Dermatol,2001,117(5):1218-1224.
[12]张丽宏,雄辉,胡中花,等.白桦脂醇对人皮肤成纤维细胞胶原蛋白及相关蛋白酶基因的调控机制研究[J].国际中医中药杂志,2016,38(5):420-423.Zhang LH,Xiong H,Hu ZH,et al.Regulative effect of collagen and related protease gene of betulin on human ESF-1 cells[J].Int J Trad Chin Med,2016,38(5):420-423.
[13]Varani J,Perone P,Deming MO,et al.Impaired keratinocyte funetion on matrix metalloproteinase-1(MMP-1)damaged collagen[J].Arch Der-matol Res,2009,(301):497-506.
[14]来吉祥,何聪芬,董银卯.皮肤衰老机理及延缓衰老化妆品的研究进展[J].中国美容医学,2009,8(75):1208-1212.Lai JX,He CF,Dong YM.Research development on the mechanism of skin aging and anti-aging cosmetics[J].Chin J Aesthet Med,2009,8(75):1208-1212.
[15]裴瑞卿.基质金属蛋白酶家族介绍(英文)[J].中国生物化学与分子生物学报,2001,17(1):1-6.Pei RQ.The matrix metalloproteinase family:An introduction[J].Chin JBiochem Mol Biol,2001,17(1):1-6.
[16]孟飞,王丽,王彦,等.17β雌二醇对人皮肤成纤维细胞胶原和弹性蛋白合成的影响[J].组织工程与重建外科杂志,2016,12(2):111-113.Men F,Wang L,Wang Y,et al.Effects of 17β-estradiol on the synthesis of collagen and elastin in cultured human skin fibroblasts[J].J Tissue Eng Reconstruct Surgery,2016,12(2):111-113.
[17]张昉,高蓉.维生素C检测方法研究进展[J].中国卫生检验杂志,2009,(2):457-460.Zhang F,Gao R.Research progress on vitamin C detection method[J].Chin J Health Inspect,2009,(2):457-460.