五味子提取工艺优化及乙醇提取物的皮肤细胞抗氧化作用研究

Study on optimized extraction of Schisandra chinensis and cutaneous cell antioxidative effect of its extract

目的探究五味子提取物对人皮肤细胞氧化损伤的保护作用,开发其在皮肤抗氧化方面的应用。方法分别采用热水蒸煮和水蒸气蒸馏的方法进行脱色和除味;另外,采用叔丁基过氧化氢(tBHP)诱导的HaCaT细胞氧化应激模型评价五味子提取物对人皮肤细胞氧化损伤的保护作用,采用CCK-8试剂盒检测细胞存活率,AnnexinV-FITC/PI双标记流式细胞仪检测细胞凋亡率,2,7-二氯荧光黄双乙酸盐法检测细胞内活性氧的水平。结果采用优化后的提取工艺得到了气味变淡且颜色明显变浅的五味子提取物;用北五味子提取物预处理HaCaT细胞6 h后,可有效降低tBHP引起的细胞凋亡,提高细胞存活率,降低细胞内ROS水平。结论优化后的乙醇提取工艺显著改善了五味子提取物的颜色和气味,并保持了提取物的抗氧化活性,因而有望成为一种应对皮肤氧化损伤的外用保护剂。

Objective To investigate the anti-oxidative effects of the Schisandra chinensis extract in skin cells and develop the application as skin anti-oxidant.Methods The ethanol extraction process of Schisandra chinensis was optimized with hot water pre-treatment and steam distillation to improve the color and odor of the extract.The tert-butyl hydroperoxide(tBHP)-induced cell oxidative injured model in HaCaT keratinocytes was used as a cellular model to study the protective effects of the extract.The cell viability was assayed with CCK-8 kit.Cell apoptosis was detected with flow cytometry by AnnexinV-FITC and PI staining.The intracellular level of reactive oxygen species(ROS) was measured by 2,7-dichlorodi-hydrofluorescein diacetate(DCFH-DA) assay.Results The Schisandra extract prepared by optimized extraction technology exhibited significant improvements in color and odor.The HaCaT cells pretreated with Schisandra extract effectively reduced the cell apoptosis induced by tBHP.The extract increased the survival rate of HaCaT cells and decreased intracellular ROS level.Conclusion The extract with optimized technology improved the color and odor.Its antioxidant activity was still maintained.It could be developed into a novel skin protectant against oxidative damage.