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
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摘要
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.
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.
引文
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[11] LEE HJ,KIM CY.Simultaneous determination of nine lignans using pressurized liquid extraction and HPLC-DAD in the fruits of Schisandra chinensis[J].Food Chem,2010,120(4):1224-1228.
[12] 金银萍,侯微,高薇,等.五味子抗氧化活性组分筛选[J].中国药房,2016,27(19):2622-2625.
[13] 蒋益萍,张巧艳,张宏,等.正交试验优选五味子木脂素类成分的提取工艺[J].中成药,2013,35(11):2390-2394.
[14] 高明清.扇贝多肽经由ROS/JNK/caspase-3通路抑制紫外线诱导的HaCaT细胞凋亡[D].青岛:青岛大学,2007.
[2] 李坦城,安丽萍,滕昊林,等.五味子提取物的研究现状与进展[J].中国医药科学,2014,4(19):80-82.
[3] 孙建晨,尹水日,崔泰花,等.北五味子提取液清除DPPH自由基的作用[J].食品科技,2012,37(6):247-250.
[4] 高翔,陈晓宇,田振坤,等.北五味子萃取物清除DPPH自由基作用研究[J].哈尔滨商业大学学报(自然科学版),2016,32(5):546-548.
[5] 李振,孟宪纬,管延珍,等.南、北五味子提取物的抑菌及抗氧化作用研究[J].中国调味品,2017,42(1):28-30.
[6] 曹波,路婷婷,魏菲,等.南五味子提取物对UVB辐射的防护作用观察[J].中国皮肤性病学杂志,2016,30(8):771-775+798.
[7] LAM PY,YAN CW,CHIU PY,et al.Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue[J].Fitoterapia,2011,82(3):393-400.
[8] 侯微,魏忠宝,高薇,等.五味子甲素·乙素及丙素对HaCat细胞氧化损伤的保护作用研究[J].安徽农业科学,2013,41(3):1047-1049.
[9] 田双,严铭铭,邵帅,等.五味子醇提物的提取工艺优化及其体外抗氧化作用评价[J].中国药房,2016,27(28):3976-3978.
[10] 李霞,贾晓斌,陈彦,等.五味子提取工艺的优化研究[J].中国药房,2007,18(6):424-426.
[11] LEE HJ,KIM CY.Simultaneous determination of nine lignans using pressurized liquid extraction and HPLC-DAD in the fruits of Schisandra chinensis[J].Food Chem,2010,120(4):1224-1228.
[12] 金银萍,侯微,高薇,等.五味子抗氧化活性组分筛选[J].中国药房,2016,27(19):2622-2625.
[13] 蒋益萍,张巧艳,张宏,等.正交试验优选五味子木脂素类成分的提取工艺[J].中成药,2013,35(11):2390-2394.
[14] 高明清.扇贝多肽经由ROS/JNK/caspase-3通路抑制紫外线诱导的HaCaT细胞凋亡[D].青岛:青岛大学,2007.