金钗石斛复方在细胞水平的美容功效研究

Study of cosmetological efficacy of Dendrobium nobile prescriptions at cell theoretical level

通过水提取法得到金钗石斛复方提取液,生药质量浓度为0.5 g·mL-1。采用XTT比色法检测金钗石斛复方提取液对表皮角质细胞的线粒体活性、观察细胞增殖并检测其对ATP含量的影响;利用多巴染色等方法,检测金钗石斛复方提取液对黑素细胞酪氨酸酶活性的抑制和黑素生成的影响;利用免疫荧光法检测金钗石斛复方提取液对角质细胞保湿相关蛋白表达的影响。实验结果表明,金钗石斛复方提取液在终质量浓度为2.5 g·L-1时,较对照组可以显著提升表皮细胞的线粒体活性和ATP含量,能够明显促进细胞的增殖。金钗石斛复方提取液在不影响共培养细胞状态的前提下,较对照组可以明显降低黑素细胞的黑色素含量并抑制酪氨酸酶活性。同时金钗石斛复方还可以促进角质细胞水通道蛋白AQP3、紧密连接蛋白ZO-1及Claudin-1的表达。

Dendrobium nobile prescriptions were extracted with distilled water and the mass concentration of Dendrobium nobile in the extract is 0. 5 g · mL^-1. Effects of Dendrobium nobile in the extract on mitochondrial activity of epidermal keratinous cell were detected with XTT colorimetry, as well as cell proliferation observed and effect on cellular ATP contents of HaCaT cell detected. Effects of Dendrobium nobile in the extract on the inhibition of activity of melanin cell tyrosinase and formation of melanin were detected with dopa staining, et al. Effects of Dendrobium nobile prescriptions in the extract on expression of moisturizing related proteins in keratinous cell HaCaT was detected with immunofluorescence method. Results showed that the extracts of Dendrobium nobile prescriptions with mass concentration of 2.5 g · L^-1 can significantly increase the mitochondrial activity of epidermal keratinous cell and cellular ATP content as comparing with that of the control group and can apparently promote proliferation of HaCaT cell. Further, melanin content in melanophore is significantly reduced and activity of tyrosinase is inhibited without affecting cell state in vitro model of the melanocyte and HaCaT cell co - culture system. Meanwhile, the extract of Dendrobium nobile prescriptions also can promote the expression of the aquaglyceroporin of keratinous cell protein AQP3, tight junctions protein ZO - 1 and Claudin - 1