摘要
Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology.In the present study,we investigated the melanogenic activity of six structurally distinct compounds,namely,scopoletin,kaempferol,chrysin,vitamin D_3,piperine,and 6-benzylaminopurine.We determined their effectiveness,toxicity,and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model.The melanogenic activity of 6-benzylaminopurine,the compound identified as the most potent,was further verified by measuring green fluorescent protein concentration in tyrp1 a:eGFP(tyrosinase-related protein 1)zebrafish and mitfa:eGFP(microphthalmia associated transcription factor)zebrafish and antioxidative activity.All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF.6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20μmol·L^(-1 )in vivo and 100μmol·L^(-1) in vitro,and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1 a:e GFP and mitfa:e GFP zebrafish in vitro.However,its relative anti-oxidative activity was found to be very low.Overall,our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism,by increasing melanin content via positive regulation of tyrosinase activity in vitro,as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.
Several chemical compounds can restore pigmentation in vitiligo through mechanisms that vary according to disease etiology.In the present study,we investigated the melanogenic activity of six structurally distinct compounds,namely,scopoletin,kaempferol,chrysin,vitamin D3,piperine,and 6-benzylaminopurine.We determined their effectiveness,toxicity,and mechanism of action for stimulating pigmentation in B16F10 melanoma cells and in a zebrafish model.The melanogenic activity of 6-benzylaminopurine,the compound identified as the most potent,was further verified by measuring green fluorescent protein concentration in tyrp1 a：eGFP（tyrosinase-related protein 1）zebrafish and mitfa：eGFP（microphthalmia associated transcription factor）zebrafish and antioxidative activity.All the tested compounds were found to enhance melanogenesis responses both in vivo and in vitro at their respective optimal concentration by increasing melanin content and expression of TYR and MITF.6-Benzyamino-purine showed the strongest re-pigmentation action at a concentration of 20μmol·L^-1 in vivo and 100μmol·L^-1 in vitro,and up-regulated the strong fluorescence expression of green fluorescent protein in tyrp1 a：e GFP and mitfa：e GFP zebrafish in vitro.However,its relative anti-oxidative activity was found to be very low.Overall,our results indicated that 6-benzylaminopurine stimulated pigmentation through a direct mechanism,by increasing melanin content via positive regulation of tyrosinase activity in vitro,as well as up-regulating the expression of the green fluorescent protein in transgenic zebrafish in vivo.
基金
supported by the National Natural Science Foundation of China(No.81874331)
the Open Project of State Key Laboratory of Natural Medicines(No.3144060130)
