细梗蔷薇愈伤组织提取物在人真皮成纤维细胞及3D表皮模型上的护肤功效探究

Study on skin care effects of Rosa graciliflora callus extract in human dermal fibroblast cells and 3D epidermal model

通过植物组织培养技术得到细梗蔷薇(Rosa graciliflora)愈伤组织,利用纯水超声提取和真空冷冻干燥工艺获得细梗蔷薇愈伤组织提取物(RCE),然后对RCE进行成分检测和体外护肤功效评估。结果表明,RCE含有氨基酸、维生素、有机酸和多酚等多种活性成分。体外护肤检测结果表明,RCE对人真皮成纤维细胞有一定的增殖作用,0.01%和0.05%RCE能促进人真皮成纤维细胞分泌Ⅰ型胶原蛋白,与空白对照对比,其Ⅰ型胶原蛋白分别提高83%和79%,作用效果和阳性对照维生素C相似。此外,在重建的3D表皮模型上,0.01%RCE能够显著促进细胞增殖活性标记物Ki67、皮肤屏障功能标志物丝聚蛋白(Filaggrin)及外膜蛋白(Involucrin)的表达,相比空白对照,其阳性率分别提高308%,853%和349%,具有显著差异性。研究发现RCE具有潜在的延缓皮肤衰老和提高皮肤屏障功能的作用,有望作为一种绿色原料添加在化妆品中使用。

The Rosa graciliflora callus was induced from seedling leaves by plant tissue culture,and the Rosa graciliflora callus extract(RCE) was obtained by ultrasonic extraction and lyophilization.To investigate the active component of RCE and its skin care efficacy in vitro,the contents of amino acids,vitamins and organic acids were detected by LC-MS,and the effect of RCE on skin was evaluated by normal human dermal fibroblast(HDF) cells and 3D epidermal model.Cells viabilities and the content of collagen I(Col-I) were detected in HDF cells to evaluate the skin anti-aging function and the immunohistochemical staining(Ki67) and immunofluorescence staining(Filaggrin and Involucrin) were detected in 3D epidermal model to evaluate the skin barrier function of RCE.The results show that 4.27%±0.23% RCE powder can be obtained by ultrasonic extraction and lyophilization.In addition,LC-MS results show that RCE contained 3.88% amino acids,0.003 5% vitamins,and 1.04% organic acids,and other experimental results show that RCE also contained 6.20% minerals,43.80% sugar,10.00% protein,1.37% polyphenols,14.6% ash and 21.90% water.Furthermore,cells assay results show that RCE can significantly promote HDF cells viability and type I collagen content.Compared with the blank control,0.01% and 0.05% RCE can increase type I collagen content by 83% and 79%,respectively(P<0.01),with the effect similar to the positive control vitamin C.Moreover,in the reconstructed 3D epidermal model assay,compared with the results of the model group,0.01% RCE can significantly up-regulate the expression of Ki67,Filaggrin and Involucrin,the relative positive rates of Ki67,Filaggrin and Involucrin are increased by 308%,853% and 349%,respectively,and the fluorescence intensities of Filaggrin and Involucrin are increased by 824% and 159%,respectively.The results indicate that RCE has the potential effect of anti-aging and improving skin barrier function and is expected to be used as a green raw material in cosmetics.