枇杷外泌体及细胞液基于体外水平的护肤功效研究  

Loquat Exosomes and Cytosol for Skin Care Efficacy in vitro

作　　者：李惠华[1] 吴美芳[1] 王伟[1] 徐夙侠[1] LI Huihua;WU Meifang;WANG Wei;XU Suxia(Fujian Institute of Subtropical Botany/Fujian Key Laboratory of Physiology and Biochemistry for Subtropical Plant,Xiamen,Fu-jian 361006,China)

机构地区：[1]福建省亚热带植物研究所/福建省亚热带植物生理生化重点实验室,福建厦门361006

出　　处：《热带作物学报》2023年第7期1488-1496,共9页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31501735);厦门市产学研项目(No.2022350204004647)。

摘　　要：探索枇杷(Eriobotrya japonica L.)外泌体及细胞液对体外人皮肤细胞的生物活性,为其在护肤品上的开发提供科学依据。采用超高速离心法和机械破碎法分别获得枇杷外泌体和细胞液,以CCK8法检测不同浓度枇杷叶来源外泌体、枇杷细胞悬浮培养来源的外泌体及其细胞液对人皮肤成纤维细胞(HSF)存活率的影响;对HSF细胞采用脂多糖(lipopolysaccharide,LPS)处理进行炎症造模,选取显著促进细胞存活率的浓度,后续开展添加外泌体或细胞液处理,进行细胞凋亡及迁移能力检测。对黑色素瘤细胞(A375)开展外泌体或细胞液处理,进行黑色素含量及酪氨酸酶活性检测。结果表明:每克枇杷叶含8.54×10^(9)个外泌体颗粒,粒径86.78 nm;每毫升(约1 g)枇杷悬浮细胞上清液中含8.67×10^(8)个外泌体颗粒,粒径80.39 nm。浓度为20μg/mL时,枇杷叶外泌体、枇杷悬浮细胞外泌体、枇杷悬浮细胞液均显著提高体外HSF细胞存活率(P<0.05)。与阳性对照(LPS处理)相比,枇杷叶外泌体浓度为20μg/mL,枇杷悬浮细胞外泌体浓度为20μg/mL和枇杷悬浮细胞液浓度为10μg/mL时,均极显著降低LPS处理引起的细胞凋亡(P<0.01),均显著促进细胞迁移(P<0.05)。浓度为5μg/mL的枇杷悬浮细胞外泌体和枇杷悬浮细胞液均可显著降低体外A375细胞黑色素含量,枇杷细胞液还显著降低酪氨酸酶活性(P<0.05)。The biological activity of loquat exosomes and cytosol on human skin cells in vitro was explored to provide a scientific basis for the development of skin care products.The loquat exosomes and cytosol were obtained by ul-tra-high-speed centrifugation and mechanical disruption,respectively.Firstly,the effect of different concentrations of loquat leaf-derived exosomes(loquat leaf-exo),loquat cell suspension culture-derived exosomes(loquat suspension-exo)and loquat cell suspension culture-derived cytosol(loquat suspension-cytosol)on cell viability of human skin fibroblasts(HSF)was detected by the CCK8 method.And then,HSF cells were treated with LPS(lipopolysaccharide)for inflam-mation modeling,and the concentration that significantly promoted the cell survival rate was selected,followed by adding exosomes or cytosolic treatment to detect cell apoptosis and migration ability.Secondly,the melanoma cells(A375)were subsequently treated with exosomes or cytosol,and the melanin content and tyrosinase activity were de-tected.The concentration of exosomes per gram of loquat leaves was 8.54×10^(9) particles,and the particle size was 86.78 nm;each milliliter(approximately equal to 1 g)of loquat suspension cells contained exosomes with a concentra-tion of 8.67×10^(8) particles,with a particle size of 80.39 nm.At the concentration level of 20μg/mL,loquat leaf-exo,lo-quat suspension-exo,and loquat suspension-cytosol all significantly increased the survival rate of HSF cells in vitro(P<0.05).Compared with the positive control(LPS treatment group),loquat leaf-exo at 20μg/mL,loquat suspen-sion-exo at 20μg/mL and loquat suspension-cytosol at 10μg/mL significantly reduced the apoptosis induced by LPS treatment(P<0.01),both significantly promoted cell migration(P<0.05).5μg/mL loquat suspension-exo and loquat suspension-cytosol could significantly reduce the melanin content of A375 cells in vitro,and loquat suspension-cytosol could also significantly reduce the tyrosinase activity(P<0.05).

关 键 词：枇杷 外泌体 悬浮细胞 细胞液 护肤功效 

分 类 号：TS209[轻工技术与工程—食品科学]