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
Objective To evaluate the anti-osteoporosis effect of the mixtures of epimedin C and icariin monomers with invariant molarity on zebrafish osteoporosis model. Methods The zebrafishes, fertilized 4 d ago, were exposed ...Objective To evaluate the anti-osteoporosis effect of the mixtures of epimedin C and icariin monomers with invariant molarity on zebrafish osteoporosis model. Methods The zebrafishes, fertilized 4 d ago, were exposed in 11 groups of nutrient solutions with prednisolone （25 μmol/L） as well as epimedin C and icariin （15 μmol/L） of various contents. The ratio of epimedin C and icariin in the 11 groups were as follows： A （10：0）, B （9：1）, C （8：2）, D （7：3）, E （6：4）, F （5：5）, G （4：6）, H （3：7）, I （2：8）, J （1：9）, and K （0：10）. Meanwhile, a negative control group with prednisolone （25μmol/L） was prepare as S. The selected zebrafish fetus was put into the 24-hole culture plate, and ensure every 5 zebrafishes in 1 hole and 2 holes as a group. They were placed in incubator at 28.5 oC, and the daily changes of fluid were investigated until they were put to death on day 8 and then fixed. After dyeing with alizarin red, the segmental venter of zebrafish skulls was observed and quantitative analysis of dyed area was conducted. Results Compared with the negative control group S, the integrated optical density （IOD） values of cranial dyed area in all groups increased significantly （P 〈 0.05）; Compared with group S, the IOD value of cranial dyed area in mixtures of epimedium monomers increased significantly （P 〈 0.05）. The mixtures of epimedium monomers were all effective in facilitating zebrafish cranial mineralization and preventing prednisolone from inducing osteoporosis. According to mixtures of A-K groups, zebrafish cranial mineralization gradually decreased with gradually reduced content of epimedin C, with significant difference among groups （P〈 0.05）. Conclusion The higher the content of epimedin C in mixtures with invariant molarity is, the more active the anti-osteoporosis effect of epimedinC to zebrafish osteoporosis model is.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
目的:用 HPLC 梯度洗脱法测定巫山淫羊藿中4种主要成分的含量,考察了淫羊藿属苷 A、双藿苷 A、朝藿定 C 和淫羊藿苷在巫山淫羊藿不同部位中的分布。方法:采用 RP-HPLC 法测定,以 BECKMAN COULTER_(TM)-C_(18)(10μm,4.6 mm×250mm)...目的:用 HPLC 梯度洗脱法测定巫山淫羊藿中4种主要成分的含量,考察了淫羊藿属苷 A、双藿苷 A、朝藿定 C 和淫羊藿苷在巫山淫羊藿不同部位中的分布。方法:采用 RP-HPLC 法测定,以 BECKMAN COULTER_(TM)-C_(18)(10μm,4.6 mm×250mm)为色谱柱,流动相为乙腈-水,梯度洗脱(0 min,乙腈-水比例为20:80;5 min,比例为30:70;10 min,比例为50:50),流速为1 mL·min^(-1),检测波长270 nm。结果:淫羊藿属苷 A 在0.188μg～1.880μg,淫羊藿苷在0.214μg～2.140μg,双藿苷 A在0.240μg～2.400μg,朝藿定 C 在0.282μg～2.820μg范围内呈良好线性关系;重复性实验的 RSD 分别为1.2%,1.3%,1.2%,1.2%。巫山淫芋藿不同部位双藿苷 A 和朝藿定 C 的含量较大,同一植株的不同部位中朝藿定 C 的分布为根>叶>茎。双藿苷 A 的分布为根>茎>叶,淫羊藿属苷 A 和淫羊藿苷含量较少。结论:巫山淫羊藿根中的化学成分以朝藿定 C 为最高,双藿苷 A 为次,而巫山淫羊藿叶中也以朝藿定 C 含量为最高。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.
基金Open Project of Key Laboratory of Plant Resources and Sustainable Utilization of Chinese Academy of SciencesProject of Health and Family Planning Commission of Wuhan Municipality(Grant No.W2014ZT288)
文摘Objective To evaluate the anti-osteoporosis effect of the mixtures of epimedin C and icariin monomers with invariant molarity on zebrafish osteoporosis model. Methods The zebrafishes, fertilized 4 d ago, were exposed in 11 groups of nutrient solutions with prednisolone （25 μmol/L） as well as epimedin C and icariin （15 μmol/L） of various contents. The ratio of epimedin C and icariin in the 11 groups were as follows： A （10：0）, B （9：1）, C （8：2）, D （7：3）, E （6：4）, F （5：5）, G （4：6）, H （3：7）, I （2：8）, J （1：9）, and K （0：10）. Meanwhile, a negative control group with prednisolone （25μmol/L） was prepare as S. The selected zebrafish fetus was put into the 24-hole culture plate, and ensure every 5 zebrafishes in 1 hole and 2 holes as a group. They were placed in incubator at 28.5 oC, and the daily changes of fluid were investigated until they were put to death on day 8 and then fixed. After dyeing with alizarin red, the segmental venter of zebrafish skulls was observed and quantitative analysis of dyed area was conducted. Results Compared with the negative control group S, the integrated optical density （IOD） values of cranial dyed area in all groups increased significantly （P 〈 0.05）; Compared with group S, the IOD value of cranial dyed area in mixtures of epimedium monomers increased significantly （P 〈 0.05）. The mixtures of epimedium monomers were all effective in facilitating zebrafish cranial mineralization and preventing prednisolone from inducing osteoporosis. According to mixtures of A-K groups, zebrafish cranial mineralization gradually decreased with gradually reduced content of epimedin C, with significant difference among groups （P〈 0.05）. Conclusion The higher the content of epimedin C in mixtures with invariant molarity is, the more active the anti-osteoporosis effect of epimedinC to zebrafish osteoporosis model is.
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。
文摘目的:用 HPLC 梯度洗脱法测定巫山淫羊藿中4种主要成分的含量,考察了淫羊藿属苷 A、双藿苷 A、朝藿定 C 和淫羊藿苷在巫山淫羊藿不同部位中的分布。方法:采用 RP-HPLC 法测定,以 BECKMAN COULTER_(TM)-C_(18)(10μm,4.6 mm×250mm)为色谱柱,流动相为乙腈-水,梯度洗脱(0 min,乙腈-水比例为20:80;5 min,比例为30:70;10 min,比例为50:50),流速为1 mL·min^(-1),检测波长270 nm。结果:淫羊藿属苷 A 在0.188μg～1.880μg,淫羊藿苷在0.214μg～2.140μg,双藿苷 A在0.240μg～2.400μg,朝藿定 C 在0.282μg～2.820μg范围内呈良好线性关系;重复性实验的 RSD 分别为1.2%,1.3%,1.2%,1.2%。巫山淫芋藿不同部位双藿苷 A 和朝藿定 C 的含量较大,同一植株的不同部位中朝藿定 C 的分布为根>叶>茎。双藿苷 A 的分布为根>茎>叶,淫羊藿属苷 A 和淫羊藿苷含量较少。结论:巫山淫羊藿根中的化学成分以朝藿定 C 为最高,双藿苷 A 为次,而巫山淫羊藿叶中也以朝藿定 C 含量为最高。
