Effect of Recipe of Supplementing Ying-wei on Expression of Wnt/β-catenin Signal Pathway Related Proteins in Epidermal Stem Cells of Aging Mice
Objective To study the effect of Recipe of Supplementing Ying-wei on the protein and mRNA ex-pression levels of Wnt/[3-catenin signal pathway including C-myc,Axin2,MMP-7,Dsh1,LEF-1 in skin epi-dermal 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 el-derly 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 com-plete culture-medium containing 160 ng·mL-1 DKK 1+2.5％drug serum concentration,the medicine was sup-plemented Ying-wei,consisting of 5 g ginseng,15 g raw astragalus,and 7 g roasted licorice,with a concen-tration of 0.5 g·mL-1.The expression levels of C-myc,Axin2,MMP-7,Dsh1 and LEF-1 protein in mice epi-dermal 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 epider-mal stem cells of the drug intervention group(P all＜0.01).Compared with the young group,the mRNA expres-sion levels of C-myc,Axin2,MMP-7,Dsh1 and LEF-1 in epidermal stem cells of aged mice were significantly higher(P＜0.05 or P＜0.01).Compared with the old group,the expression levels of C-myc,Axin2 and LEF-1 mRNA in mice epidermal stem cells of the blocker group reduced significantly(P＜0.05 or P＜0.01),but the expression levels of MMP-7 and Dsh1 mRNA were not statistically significant(P all＞0.05).Compared with the old group,the expression levels of the above five mRNA reduced significantly in mice epidermal stem cells of the blocker group(P＜0.05 or P＜0.01).Compared with the blocker group,the mRNA levels of C-myc,Axin2,Dsh1 and LEF-1 in epidermal stem cells in the drug intervention group reduced significantly(P＜0.05orP＜0.01),but the expression of MMP-7 mRNA was not statistically significant(P＞0.05).Conclusion Recipe of Sup-plementing Ying-wei could significantly reduce the expression of C-myc,Axin2,MMP-7,Dsh1,LEF-1 protein and C-myc,Axin2,Dsh1,LEF-1 mRNA in Wnt/β-catenin signal pathway in aging mice epidermal stem cells and has synergistic effect with its blocker DKK1 in reducing the activity of Wnt/β-catenin signal pathway to delay skin aging.
recipe of supplementing Ying-weiWnt/β-catenin signal pathwayepidermal stem cellsskin ag-ing
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维普
