Study On The Mechanism Of Melanin Degradation Based On Lysosomal Pathway And The Intervention Of Isoglycyrrhizin And Isoimperatorin

Objective: Explore the relevance of lysosome and melanin degradation;to observe the effects of isoliquiritigenin,isoimperatorin intervention activity of lysosomal enzyme and plain black degradation.Methods:1.The establishment of co culture of melanocytes and Ha Ca T cells:A375 melanoma cells and Ha Ca T human keratinocytes were co cultured with Transwell cells.2.Determination of the safety concentration of glycyrrhizin and isoimperatorin: Ha Ca T cells were treated with different concentrations of different concentrations of different levels of glycyrrhizin and MTT to determine the proliferation rate of Ha Ca T cells.3.Determination of melanin content in Ha Ca T cells by the treatment of different kinds of drugs with different drugs:Using Na OH lysis method on Haca T cell lysis,analysisde lysate A405 value.4.The recognition of lysosomal enzymes involved in the degradation of melanin and Effect of glycyrrhizin and isoimperatorin on lysosomal enzyme activity :Using Na OH lysis method for lysis of Haca T,using the kit of lysosomal acid phosphatase associated hydrolase activity were analyzed.By analyzing the relationship between the changes of the activity of lysosomal enzymes and melanin content changes.Results:Isoliquiritigenin,isoimperatorin the maximum safe concentration of 8ug/ml,determination of OD value of MTT method,different concentrations of isoliquiritigenin,isoimperatorin Hu Sugan calculated the rate of cell proliferation and prognosis,two have promoting effect on cell proliferation at the concentration of 0.5ug/ml and therefore excluded in isoliquiritigenin,isoimperatorin 2ug/ml.4ug/ml,8ug/ml cell proliferation rate was more than 90%,and had no significant effect on cell proliferation.Isoliquiritigenin,isoimperatorin intervention melanin content in cells decreased gradually,and has a certain concentration dependence.Isoliquiritigenin,isoimperatorin on acid enzyme activity of acidic phosphorus promoted but,the effect does not have concentration dependent,and in 4ug/ml on the activity of the greatest influence.4ug/ml is selected for subsequent experiments,adding the lysosomal inhibitor,increased melanin content in the cell,adding isoliquiritigenin,isoimperatorin intracellular melanin content decreased.Conclusion:Lysosomal hydrolases may be involved in keratinocyte cell degradation of melanin,isoliquiritigenin,isoimperatorin Hu Su change intervention to promote activity of lysosomal enzymes,thereby promoting degradation of melanin,and the two optimal concentration of 4ug/ml.