Influence Of Phenylethanoid Glycosides From Cistanche On Melanocytes Melanin Synthesis And Its Mechanism

Objective:1.The interference of phenylethanoid glycosides from Cistanche in the metabolism of melanocytes was evaluated,including the effects the proliferation of cells,tyrosinase activity and melanin content.2.Screening of radiation dose of UVB,establishing the model of UVB radiation induced pigmentation.3.To establish the model of pigmentation cells,and to explore the influence of pigmentation phenylethanoid glycosides on UVB radiation;4.To investigate the inhibitory effects of phenylethanoid glycosides from cistanche on melanogenesis in cultured human epidermal melanocytes,and explore the structure-activity relationship through comparison of the inhibition effect of ten kinds of PhGs.Method:MTT assay after different intensities of cell proliferation rate of the cells,L-DOPA assay cells tyrosinase activity,NaOH lysis assay melanin content,RT-PCR assay related protein MITF,TRY,TRP-1,TRP-2 expression.From the cellular level and molecular level detection phenylethanoid glycosides from Cistanche on melanocytes melanin synthesis and its mechanism.Results:1.phenylethanoid glycosides from Cistanche on melanocytes tyrosinase activity significantly inhibited,effectively inhibit the proliferation of melanocytes in a dose-response relationship,melanocytes also significantly reduced melanin content,along with phenylethanoid glycosides from Cistanche increasing the total glycoside concentration adjusted G₁/S transition and induces cell apoptosis.400?g/mL phenylethanoid total glycosides from Cistanche can be significantly reduced TYR,TRP-1,TRP-2 and MITF mRNA expression levels.2.0.3J/cm² UVB model group compared with the control group,the proliferation rate of cell proliferation rate was significantly higher?P<0.01?,0.3 J/cm² UVB model group and control group,tyrosinase activity significantly increased,the difference was statistics significance?P<0.05?,cell melanin content significantly increasing trend,and the difference was statistically significant?P<0.01?.After 0.3 J/cm² UVB intervention can significantly increase cell MITF,TRY,TRP-1,TRP-2 mRNA.3.The study proved that phenylethanoid glycosides from Cistanche obviously inhibit the melanin biogenesis of melanocytes promoted by UVB.phenylethanoid glycosides from Cistanche can effectively inhibit melanocytes tyrosinase activity and melanin content,cell apoptosis with flow cytometry results show that phenylethanoid glycosides in different concentrations in the intervention of UVB induced pigmentation effect concentration is safe.Phenylethanoid glycosides significantly reduced melanin synthesis related TYR?TRP-1 and MITF mRNA expression levels,200?g/ml and 400?g/ml phenylethanoid glycosides was significantly downregulated in Melanin synthesis related genes of TYR?TRP-1?TRP-2 and MITF protein expression levels.4.PhGs reduced tyrosinase activity in a dosed-depended manner.The content of melanin was reduced by PhGs.Compared with the positive control,significant elevation was noticed in the inhibitory effect of poliumoside,2'-acetylacteoside,acteoside,oleuropein and calceolarioside B?P<0.01?,respectively.On the contrary,the inhibitory effects of forsythoside A/B was relatively weaker compared with positive control group?P<0.01?.Conclusion:Phenylethanoid glycosides from Cistanche by inhibition of cell proliferation and tyrosinase activity decreased and Melanin synthesis decline,also regulation Melanin synthesis related gene and protein expression,thereby reducing the synthesis of melanin in.UVB radiation has a positive regulatory effect on the expression of related genes in the pathway of melanin synthesis.Phenylethanoid glycosides could significantly inhibit after UVB radiation,tyrosinase activity and melanin content,molecular level down melanin synthesis related gene and protein expression.Phenylethanoid glycosides in the intervention of UVB induced pigmentation of normal cell growth were not significantly affected.Therefore,phenylethanoid glycosides can be added to the whitening and anti-aging cosmetics,but the definite,practical effect is also affected by preparation of carrier effect,still must strictly for in vivo animal experiments and human skin validity test.In this study,,To investigate the inhibitory effects of phenylethanoid glycosides from cistanche on melanogenesis in cultured human epidermal melanocytes,and explore the structure-activity relationship through comparison of the inhibition effect of ten kinds of PhGs.beneficial to discover new restrain skin pigment calm drug or chemical synthesis provide lead compounds,in order to develop new drugs with independent intellectual property rights.