Experimental Study Of Molecular Mechanism Of Vernoniaanthelmintica Willd Treating Vitiligo

Vitiligo is characterized by depigmentation in large or small areas of the skin. There is a complete lack of melanin production in the hypopigmented areas, due to the destruction of melanocytes. Although the etiology of vitiligo remains unknown, an autoimmune theory is gaining acceptance. In order to provide the experimental basis for the detailed molecular immunological mechanism of vitiligo and clinical use of vemoniaanthelrnintica willd injuction, we have explored the effect of the Vernoniaanthelmintica willd injection (traditional Chinese medicine) on the immune function of mice, tyrosinase activity, melanoma cells proliferation , the melanogenesis and expression of tyrosinase gene in of A375 human melanoma cells, finaly identified peptide mass fingerprinting of the vitiligo-related proteins.1 Effects of vernoniaanthelmintica willd on the immune function of miceWe have identified the action of vernoniaanthelmintica willd injection (a traditional Chinese medicine) on the immune function of mice, using [3H]-TdR incorperation , spleen cell mediated sheep red blood cell quantitative hemolytic spectrophotometry, delayed type of hyper-sensitivity (DTH) and flow cytometer methods respectively. Results showed vemoniaanthelrnintica willd injection could inhibit spleen T.B lymphocyte proliferation of normal mouse in vitro and in vivo, inhibit antibody forming and DTH in vivo, Vemoniaanthehnintica willd could also inhibit spleen T lymphocyte cecretion of IL-2, the total immunoglobulin, specific antibody against A3 75 human melanocytoma cell and B cellsurface antigen (CD 19) express of normal mouse in vivo. In a word vernoniaanthelmintica willd injection could inhibit the cell mediated immune function and humoral immune function of normal mouse.2 The studes of Effect of Vernoniaanthelmintica Willd on tyrosinase activityWe have studied the effect of Vernoniaanthehnintica Willd on tyrosinase activity , the melanoma cell proliferation and the melanogenesis of A3 75 human and B16 murine melanoma cell ,by zymologic method, MTT methods. Results showed vernoniaanthelmintica willd injection could significantly activate the tyrosinase activity in vitro, and in vivo, could enhance the melanoma cells proliferation and the melanogenesis of A375 human melanoma cell in vitro.3 Effect of Vernoniaanthehnintica Willd on Gene Expression of Tyrosinase of A375 Human Melanoma cellsOn the basis of above,we have studied the effect of vernoniaanthelmintica willd on geneticexpression of tyrosinase of A375 human melanoma cell, we first purified tatol RNA by TRIzol kit from cultured A3 75 human melanoma cells, the expression level of tyrosinase mRNA were assessed by RT-PCR technique. Results showed that vemoniaanthelmintica willd significantly can enhance the expression of tyrosinase mRNA in cultured A3 75 human melanoma cells. It is proved that vemoniaanthelmintica willd can enhance the synthesis of tyrosinase from gene level.4 A mass spectrometric analysis of A375 human melanoma cell proteinsA molecular weight 45kD vitiligo-relatived protein were identified by western blotting, where the A375 human melanoma cell proteins was resolved by SDS-polyacrylamide gel electrophoresis , transfer to nitric acid cellulose (NC) membrane and to react with vitiligo patient serum, according to report 40kD and 90kD also were vitiligo-relatived proteins, the peptide mass fingerprinting(PMF) of the three proteins were identified by MALDI-TOP-MS and showed these proteins were new proteins.