Experiental Studies Of Non-photosensitive Traditional Chinese Medicinal Herbs Of Its Effects On Vitiligo

Objective Through animal experiments,to explore the possible mechanism of extracts of non-photosensitive traditional Chinese Medicinal Herbs(No.2 Traditional Chinese Medicinal Herbs,No.2 TCMHs.composed by Glossy Privet Fruit,Balck Pepper,Duckweed,Tuberculate Speranskia Herb,Stenoloma chusanum Ching) in the treatment of vitiligo.Through cell experiments,in order to explore its possible mechanism of tyrosol and piperine on the proliferation and/or melanogenesis of murine B 16 melanoma cells.Methods(1)Animal experiments- No.2TCMHs were extracted by alcohol,then the rat skin was treated by the ethanol extracts twice a day,and ethanol was used as negative control.Four weeks later,the skin specimens were obtained. Immunohistochemistry method was used to detect the S100,Kit-tyrosine kinase receptor(c-KIT) and proteinase-activated receptor-2(PAR-2) protein expression in these specimens.(2) Cell Experiments:After treatment of murine B 16 melanoma cells with either tyrosol or piperine for 48h or 72h,the cell proliferation was detected by MTT,the tyrosinase activity by DOPA oxidation assay,the melanin content by NaOH-assay,and the expression of c-KIT and TRP-2 protein were measured by immunohistochemistry.Results(1)Compared with the ethanol group,there was a significant increase in the positive rates of c-KIT and PAR-2 expression in No.2TCMHs treated group(P＜0.05), while there was no significant increase of the S100 positive cells(P＞0.05).(2) Tyrosine activity and melanogenesis were increased with tyrosol of the concentration of 25μg/mL(P＜0.05),and the protein expression of c-KIT and TRP-2 were also increased by 9.50%(P＜0.05) and 9.16%(P＜0.01),respectively.Cell proliferation (P＜0.05) and tyrosine activity(P＜0.01) as well as melanogenesis(P＜0.05) were increased with piperine of the concentration of 12.5μg/mL,and the protein expression of c-KIT and TRP-2 were also increased by 8.81%(P＜0.05) and 10.58% (P＜0.05),respectively. Conclusions(1) Our results suggest that the efficacy of non-photosensitive traditional Chinese Medicinal Herbs in the treatment of vitiligo may be related to its ability to enhance cell migration and melanin transfer,and not related to the increasing of melanocytes.(2) Both tyrosol and piperine could stimulate melanogenesis in murine B16 melanoma cells,which may partly be induced by upregulating of tyrosinase activity as well as expression of c-KIT and TRP-2 protein.(3) Our results raised new ideas for how to composing Traditional Chinese Recipes for Vitiligo,and provided more experimental basis for choosing Drugs of Traditional Chinese Recipes for Vitiligo.