构树叶黄酮类物质提取工艺的优化及其对小鼠表皮干细胞的抗氧化作用  

Optimization of extraction process of flavonoids from Broussonetia papyrifera leaves and its antioxidant effect on mouse epidermal stem cells

作　　者：岑丽航 孙鸽云 王珍珍 肖瑞琳 王丽丽[3] 常冰梅 CEN Lihang;SUN Geyun;WANG Zhenzhen;XIAO Ruilin;WANG Lili;CHANG Bingmei(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;不详)

机构地区：[1]山西医科大学基础医学院生物化学与分子生物学教研室,山西太原030001 [2]太原市铁路局疾病预防控制中心,山西太原030001 [3]山西医科大学第一医院,山西太原030001

出　　处：《中国生物制品学杂志》2024年第3期343-349,355,共8页Chinese Journal of Biologicals

基　　金：山西省重点研究开发计划(201903D321101);山西省应用基础研究项目(201801D121310,201701D221254)。

摘　　要：目的优化构树叶黄酮类物质的提取工艺,并探讨黄酮类物质对小鼠表皮干细胞的抗氧化作用。方法采用单因素试验优化构树叶黄酮类物质提取工艺中的液料比(15∶1、20∶1、25∶1、30∶1、35∶1)、NaOH浓度(0.2%、0.4%、0.6%、0.8%、1.0%)、pH(2.5、3.0、3.5、4.0、4.5)、浸提温度(60、65、70、75、80℃)。在单因素试验结果的基础上,以黄酮类物质质量分数为评价指标,采用正交试验设计确定最佳提取工艺。免疫磁珠法分选CD49f+/CD71-小鼠表皮干细胞,并检测黄酮类物质对其细胞相对活力及细胞中还原型谷胱甘肽(reduced glutathione,GSH)和丙二醛(malondialdehyde,MDA)含量的影响。结果黄酮类物质提取工艺的最佳条件为:液料比30∶1、NaOH浓度0.6%、pH4.5、浸提温度75℃。最佳条件下提取黄酮类物质平均质量分数为1.47%。与阴性对照组比较,当黄酮类物质终浓度为25及50μg/mL时,细胞相对活力明显增加(F分别为1.427和13.747,P均<0.01);当黄酮类物质终浓度为12.5、25、50μg/mL时,GSH含量明显上升(F分别为0.044、0.291和2.577,P均<0.05),MDA含量明显下降(F分别为3.568、4.909和1.400,P均<0.05)。结论优化后的构树叶黄酮类物质提取工艺稳定可靠,有利于构树叶加工后剩余原液的再利用,提取的黄酮类物质可促进小鼠表皮干细胞增殖,具有抗氧化性。Objective To optimize the extraction process of flavonoids from Broussonetia papyrifera leaves and explore the antioxidant effect of flavonoids on mouse epidermal stem cells.Methods The extraction process of flavonoids from Broussonetia papyrifera leaves was optimized by single factor experiment,including the liquid-solid ratio(15:1,20:1,25:1,30:1and 35:1),sodium hydroxide(NaOH)concentration(0.2%,0.4%,0.6%,0.8%and 1.0%),pH value(2.5,3.0,3.5,4.0and 4.5)and extraction temperature(60,65,70,75 and 80℃).Based on the results of single factor experiment,the optimal extraction process was determined by orthogonal test with the mass fraction of flavonoids as the evaluation index.CD49f~+/CD71~-mouse epidermal stem cells were isolated and cultivated by immunomagnetic bead method,and the effects of flavonoids on the cell relative viability and the contents of reduced glutathione(GSH)and malondialdehyde(MDA)were detected.Results The optimal extraction conditions of flavonoids were liquid-solid ratio of 30:1,0.6%NaOH,pH 4.5and extraction temperature of 75℃.Under these conditions,the average mass fraction of flavonoids extracted was 1.47%.Compared with the negative control group,when the flavonoids final concentration was 25 and 50μg/mL,the cell relative viability increased significantly(F=1.427 and 13.747 respectively,each P<0.01);when the final concentration of flavonoids was 12.5,25 and 50μg/mL,the content of GSH increased significantly(F=0.044,0.291 and 2.577 respectively,each P<0.05)and the content of MDA decreased significantly(F=3.568,4.909 and 1.400 respectively,each P<0.05).Conclusion The optimized extraction process of flavonoids from B.papyrifera leaves was stable and reliable,which is beneficial to the reuse of remaining stock solution after processing,and the extracted flavonoids can promote the proliferation of mouse epidermal stem cells and perform antioxidant activity.

关 键 词：构树叶 黄酮 表皮干细胞 抗氧化 

分 类 号：TQ914.3[化学工程]