Promotion Of Melanogenesis Of Qibai Mixture And Osthole Et Al And Their Mechanism

Objective: To study the effects of serum containing Qibai mixture on cellular proliferation, tyrosinase activity and melanin contents of cultured normal human melanocytes in vitro.And in the second part of the article,we seleceted six kinds of Chinese medicine herbal compounds, in order to obtain potential vitiligo curing agents from traditional Chinese herbs.we tested changes of melanin content in cultured human epidermal melanocyets system after treatment with Psoralen, Osthole, Paeoniflorin, Baicalin, AstragalosideⅣ, Menthol.Methods: In using the method of serum-pharmacology of traditional Chinese medicine, serum samples of rabbits containing Qibai-mixture which collected in different time(30min,60min,120min,180min,300min) were added into cultured human epidermal melanocyets(HEMn) system in vitro to estimate the cellular proliferation, tyrosinase activity and melanogenesis. The expression of TYR and MITF mRNA was examined with semiquantitative RT-PCR analysis. Psoralen, Osthole, Paeoniflorin, Baicalin, AstragalosideⅣ, Menthol with various concentration (2．5μg/ml,5μg/ml～100μg/ml) were in melanocytes system in vitro.Melanin content and cell viability were the two indications for evaluation. Tyrosinase activity and the expression of melanin synthesisrelated enzymes in cells treated with the compounds were also tested. Quantitative real-time PCR (qRT-PCR) and western-blot were used to analysis TYR and TRP-1 mRNA, protein expression.Results: Serum containing Qibai-mixture can stimulate the proliferation of melanocytes,up-regulate activation of tyrosinase and melanin contains compared with rabbit serum without drug (p<0．05) ,expecial serum sample collected with 60min,120min.serum collected of different time show different action. The level of TYR and MITF mRNA expression is significantly higher than the group control.Most of the compounds show to inhibit the proliferation of HEMn cells with high concentration(>50μg/ml).But in 5-20μg/ml, the proliferation , tyrosinase activity and melanin contents of HEMn cells were increased with treated by Psoralen, Osthole, Paeoniflorin, Baicalin. Western blot analysis revealed that expression of tyrosinase and tyrosinase-related proteins-1(TRP-1) was increased in Psoralen, Osthole, Paeoniflorin, Baicalin treated HEMn cells. Additionally, quantitative real-time PCR (qRT-PCR) analysis disclosed that expression of mRNAs for tyrosinase and TRP-1 was also enhanced by these compounds,and similar to that of 8-mop.Conclusions: These results demonstrate thet Qibai-mixture can direct effect melanocyte and tyrosinase signal pathway to treatment vitiligo or other depigmental diseases. Six compounds were proved to have repigmentation activity similar to or better than that of 8-mop and low cytotoxicity to melanocytes. Most of the compounds enhenced tyrosinase activity and the expression of tyrosinase. Four of them also promoted tyrosinase in cultured cells. These compounds in promoting pigmentation and that this action is exerted through promotion of transcription of the genes encoding tyrosinase and TRP1．...