补益营卫方对衰老小鼠表皮干细胞Wnt/β-catenin信号通路相关蛋白表达的影响  

Effect of Recipe of Supplementing Ying-wei on Expression of Wnt/β-catenin Signal Pathway Related Proteins in Epidermal Stem Cells of Aging Mice

作　　者：冯仰磊 周小平[1] 李婷 袁浩棓 FENG Yanglei;ZHOU Xiaoping;LI Ting;YUAN Haobei(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,银川750004

出　　处：《宁夏医科大学学报》2024年第3期265-270,共6页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81774318)。

摘　　要：目的探究补益营卫方对衰老小鼠表皮干细胞经Dickkopf-1(DKK1)阻断后Wnt/β-catenin信号通路中的原癌基因转录因子(C-myc)、轴抑制因子2(axis inhibitor2,Axin2)、基质金属蛋白酶-7(matrix metalloproteinase-7,MMP-7)、散乱蛋白1(dishevelled1,Dsh1)、淋巴增强因子-1(lymphoid enhancer factor-1,LEF-1)蛋白和mRNA表达水平的影响及其延缓皮肤衰老的作用。方法选取3月龄SPF级SAMR1雄性小鼠3只,作为年轻常规组;8月龄SPF级SAMP8雄性小鼠9只,随机分成衰老常规组、衰老阻断剂组和衰老药物干预组,每组3只,剪取各组小鼠的背部皮肤,分离得到表皮干细胞,并传代培养至融合。年轻常规组小鼠表皮干细胞用常规培养基培养,衰老常规组用常规培养基培养,衰老阻断剂组用含160 ng·mL^(-1)DKK1的完全培养基培养,衰老药物干预组用含160 ng·mL^(-1)DKK1+2.5%药物血清浓度的完全培养基培养(药物为补益营卫方,其组成为人参5 g、生黄芪15 g和炙甘草7 g,其浓度为0.5 g·mL^(-1))。使用Western blot法检测小鼠表皮干细胞中C-myc、Axin2、MMP-7、Dsh1、LEF-1蛋白的表达水平,RT-qPCR法检测小鼠表皮干细胞中C-myc、Axin2、MMP-7、Dsh1、LEF-1 mRNA的表达水平。结果与年轻常规组相比,衰老常规组小鼠表皮干细胞中的Cmyc、Axin2、MMP-7、Dsh1、LEF-1蛋白表达水平均升高(P均<0.01);与衰老常规组相比,衰老阻断剂组小鼠表皮干细胞中的上述5种蛋白的表达水平均降低(P均<0.01);与衰老常规组和衰老阻断剂组相比,衰老药物干预组5种蛋白表达水平均降低(P均<0.01)。与年轻常规组相比,衰老常规组小鼠表皮干细胞中的C-myc、Axin2、MMP-7、Dsh1、LEF-1的mRNA表达水平均升高(P<0.05或P<0.01);与衰老常规组相比,衰老阻断剂组小鼠表皮干细胞中的C-myc、Axin2、LEF-1 mRNA表达水平均降低(P<0.05或P<0.01),但MMP-7、Dsh1 mRNA表达差异均无统计学意义(P均>0.05);与衰老常规组相比,衰老药物干预组小�Objective To study the effect of Recipe of Supplementing Ying-wei on the protein and mRNA expression levels of Wnt/β-catenin signal pathway including C-myc,Axin2,MMP-7,Dsh1,LEF-1 in skin epidermal stem cells in aging mice blocked the by Dickkopf-1(DKK1),and the effect of delaying skin aging of Recipe of Supplementing Ying-wei.Methods Ten 3-month-old SPF grade SAMR1 male mice were selected as the young routine group;nine 8-month-old SPF grade SAMP8 male mice were randomly divided into an elderly routine group,an aging blocker group,and an aging drug intervention group,with three mice in each group.The back skin of mice of young routine group and aging age-group were cut,and epidermal stem cells were isolated and subcultured to fusion.The epidermal stem cells of young group mice were cultured in routine culture-medium,the old group were cultured in routine culture-medium,the blocker group was cultured in complete culture-medium containing 160 ng·mL^(-1) DKK1,and the drug intervention group was cultured in complete culture-medium containing 160 ng·mL^(-1) DKK1+2.5%drug serum concentration,the medicine was supplemented Ying-wei,consisting of 5 g ginseng,15 g raw astragalus,and 7 g roasted licorice,with a concentration of 0.5 g·mL^(-1).The expression levels of C-myc,Axin2,MMP-7,Dsh1 and LEF-1 protein in mice epidermal stem cells were tested by Western blot,and the expression levels of C-myc,Axin2,MMP-7,Dsh1 and LEF-1 mRNA in mice epidermal stem cells were tested by RT-qPCR.Results Compared with the young group,the protein expression levels of C-myc,Axin2,MMP-7,Dsh1 and LEF-1 in epidermal stem cells of the old group were significantly higher(P all<0.01).Compared with the old group,the expression levels of the above five proteins in mice epidermal stem cells of the blocker group were significantly reduced(P all<0.01).Compared with the old group and the blocker group,the expression levels of five proteins reduced significantly in epidermal stem cells of the drug intervention group(P all<0.01).Compared with the young

关 键 词：补益营卫方 WNT/Β-CATENIN信号通路 表皮干细胞 皮肤衰老 

分 类 号：R96[医药卫生—药理学]