Epimedin B(EB)is one of the main flavonoid ingredients present in Epimedium brevicornum Maxim.,a traditional herb widely used in China.Our previous study showed that EB was a stronger inducer of melanogenesis and an a...Epimedin B(EB)is one of the main flavonoid ingredients present in Epimedium brevicornum Maxim.,a traditional herb widely used in China.Our previous study showed that EB was a stronger inducer of melanogenesis and an activator of tyrosinase(TYR).However,the role of EB in melanogenesis and the mechanism underlying the regulation remain unclear.Herein,as an extension to our previous investigation,we provide comprehensive evidence of EB-induced pigmentation in vivo and in vitro and elucidate the melanogenesis mechanism by assessing its effects on the TYR family of proteins(TYRs)in terms of expression,activity,and stability.The results showed that EB increased TYRs expression through microphthalmia-associated transcription factor-mediated p-Akt(referred to as protein kinase B(PKB))/glycogen synthase kinase 3β(GSK3β)/β-catenin,p-p70 S6 kinase cascades,and protein 38(p38)/mitogen-activated protein(MAP)kinase(MAPK)and extracellular regulated protein kinases(ERK)/MAPK pathways,after which EB increased the number of melanosomes and promoted their maturation for melanogenesis in melanoma cells and human primary melanocytes/skin tissues.Furthermore,EB exerted repigmentation by stimulating TYR activity in hydroquinone-and N-phenylthiourea-induced TYR inhibitive models,including melanoma cells,zebrafish,and mice.Finally,EB ameliorated monobenzone-induced depigmentation in vitro and in vivo through the enhancement of TYRs stability by inhibiting TYR misfolding,TYR-related protein 1 formation,and retention in the endoplasmic reticulum and then by downregulating the ubiquitination and proteolysis processes.These data conclude that EB can target TYRs and alter their expression,activity,and stability,thus stimulating their pigmentation function,which might provide a novel rational strategy for hypopigmentation treatment in the pharmaceutical and cosmetic industries.展开更多
目的:建立同时测定淫羊藿总黄酮胶囊中朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、宝霍苷I等5种成分含量的高效液相色谱(HPLC)法。方法:所采用的色谱条件为:色谱柱:Eclipse Plus C18(250 mm×4.6 mm,5μm);流动相:乙腈-水,梯度洗脱;体...目的:建立同时测定淫羊藿总黄酮胶囊中朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、宝霍苷I等5种成分含量的高效液相色谱(HPLC)法。方法:所采用的色谱条件为:色谱柱:Eclipse Plus C18(250 mm×4.6 mm,5μm);流动相:乙腈-水,梯度洗脱;体积流量:1.0 m L·min-1;检测波长:270 nm;柱温:25℃。结果:朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、宝霍苷I分别在3.10-62.00、5.70-114.00、9.14-182.80、15.20-304.00、1.56-31.20μg·m L-1范围内呈良好的线性关系,相关系数r均大于0.999 3;平均加样回收率分别为101.06%(RSD=1.05%,n=6)、100.78%(RSD=1.08%,n=6)、99.17%(RSD=1.14%,n=6)、100.23%(RSD=0.68%,n=6)、99.09%(RSD=1.30%,n=6),该方法的精密度、重复性和稳定性良好。结论:该方法简便、准确,为淫羊藿总黄酮胶囊多成分含量测定提供一定的参考价值。展开更多
基金supported by the Open Project of National Major Science and Technology Infrastructure of Translational Medicine,China(Grant No.:TMSK-2021−404)the SIMM-SHUTCM Joint Innovation Research Program,China(2022)+1 种基金the Youth Innovation Research Foundation,China(Grant No.:A1-U21-205-01010109)the National Natural Science Foundation of China(Grant No.:81972932).
文摘Epimedin B(EB)is one of the main flavonoid ingredients present in Epimedium brevicornum Maxim.,a traditional herb widely used in China.Our previous study showed that EB was a stronger inducer of melanogenesis and an activator of tyrosinase(TYR).However,the role of EB in melanogenesis and the mechanism underlying the regulation remain unclear.Herein,as an extension to our previous investigation,we provide comprehensive evidence of EB-induced pigmentation in vivo and in vitro and elucidate the melanogenesis mechanism by assessing its effects on the TYR family of proteins(TYRs)in terms of expression,activity,and stability.The results showed that EB increased TYRs expression through microphthalmia-associated transcription factor-mediated p-Akt(referred to as protein kinase B(PKB))/glycogen synthase kinase 3β(GSK3β)/β-catenin,p-p70 S6 kinase cascades,and protein 38(p38)/mitogen-activated protein(MAP)kinase(MAPK)and extracellular regulated protein kinases(ERK)/MAPK pathways,after which EB increased the number of melanosomes and promoted their maturation for melanogenesis in melanoma cells and human primary melanocytes/skin tissues.Furthermore,EB exerted repigmentation by stimulating TYR activity in hydroquinone-and N-phenylthiourea-induced TYR inhibitive models,including melanoma cells,zebrafish,and mice.Finally,EB ameliorated monobenzone-induced depigmentation in vitro and in vivo through the enhancement of TYRs stability by inhibiting TYR misfolding,TYR-related protein 1 formation,and retention in the endoplasmic reticulum and then by downregulating the ubiquitination and proteolysis processes.These data conclude that EB can target TYRs and alter their expression,activity,and stability,thus stimulating their pigmentation function,which might provide a novel rational strategy for hypopigmentation treatment in the pharmaceutical and cosmetic industries.
文摘目的:建立同时测定淫羊藿总黄酮胶囊中朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、宝霍苷I等5种成分含量的高效液相色谱(HPLC)法。方法:所采用的色谱条件为:色谱柱:Eclipse Plus C18(250 mm×4.6 mm,5μm);流动相:乙腈-水,梯度洗脱;体积流量:1.0 m L·min-1;检测波长:270 nm;柱温:25℃。结果:朝霍定A、朝霍定B、朝霍定C、淫羊藿苷、宝霍苷I分别在3.10-62.00、5.70-114.00、9.14-182.80、15.20-304.00、1.56-31.20μg·m L-1范围内呈良好的线性关系,相关系数r均大于0.999 3;平均加样回收率分别为101.06%(RSD=1.05%,n=6)、100.78%(RSD=1.08%,n=6)、99.17%(RSD=1.14%,n=6)、100.23%(RSD=0.68%,n=6)、99.09%(RSD=1.30%,n=6),该方法的精密度、重复性和稳定性良好。结论:该方法简便、准确,为淫羊藿总黄酮胶囊多成分含量测定提供一定的参考价值。
