The Effects And Molecular Mechamism Of Medicated Blood Serum Of Baojing Oral Liquid On Melanocytes And Melanosomes Transfer

Objective:In this study,normal human melanocytes system and coculture system of epidermal keratinocytes and melanocytes were established in vitro.serum pharmacological methods were used to explore the effect and mechanism of Baojing Oral Liquid on melanin synthesis and melanosomes transfer.Methods:Preparation of medicine Serum by Serum Pharmacology.In the normal human melanocytes system,The proliferation of melanocytes?the tyrosinase activety and melanin synthesis were measured by MTT,dopaoxidase and NaOH pyrolysis methods respectively,The expression of TRYmRNA was measured by RT-PCR.Then compared the results.In the coculture system of epidermal keratinocytes and melanocytes,The expression of RhoA and Racl protein in melanocytes was detected by pull-down method,and the melanogenesis was detected by flow cytometry.Vitamin C was used as the control group.SPSS 16.0 software was used to analyze the experimental data.Results:1 Compared with the Nacl group,the effect of Baojing Oral Liquid Group and Vitamin C Group on the proliferation of melanocytes was not significantly(P>0.05).There was no significant difference between the two groups(P>0.05).Compared with the Nacl group,the medicated blood serums could decrease the melanogenesis(P<0.05).and the difference between the two groups was not statistically(P>0.05).Compared with the Nacl group,the tyrosinase activety in the medicated blood serums was decreased(P<0.05),and the difference was statistically significant(P<0.05).But there were no significant differences between the two groups(P>0.05).PCR was used to detect tyrosinase mRNA.There was no significant difference in the expression of tyrosinase mRNA between the three groups.2 The expression of RhoA and Racl protein was measured by pull-down method.Compared with the Nacl group,the expression of RhoA protein was up-regulated in the Baojing Oral Liquid group,and the expression of RhoA protein in the vitamin C group was statistically significant(P<0.05).The expression of Racl protein was down-regulated by Baojing Oral Liquid and Vitamin,and the difference was statistically significant compared with that of saline.The effect of Baojing Oral Liquid was more obvious than that of Vitamin C(P<0.05)The results showed that the transport rate of melanin was 10.32%and 20.09%,respectively,which was significantly lower than that of the Nacl group(29.43%).Baojing oral liquid group was the most significant.Conclusion:Baojing oral liquid could inhibit the activity of tyrosinase in melanocytes,reduce the synthesis of melanin.It could alao up-regulate the expression of RhoA protein,and down-regulate the expression of Racl protein,then inhibits the growth of melanocyte dendrites,reducing the melanosomes transfer.Compared with vitamin C,the inhibition of melanin synthesis was equivalent.Although Baojing Oral Liquid and Vitamin C could inhibit the expression of Racl protein,the effect of Baojing Oral Liquid was more significant than that of vitamin C.Baojing oral liquid could promote the expression of RhoA protein.Vitamin C could inhibit the expression of RhoA protein.Baojing oral liquid is better than vitamin C on the growth of melanocytes dendritic.It has important value to chloasma treatment.