Effects Of Ginger Essential Oil On Melanogenesis In Melanoma Cells And Its Mechanism

In the study,ginger essential oil was extracted by supercritical CO2 fluid.And murine B16 melanoma cells were cultured with 20-100 ?g/mL of ginger essential oil for 24 h and 48 h.The effects of ginger essential oil on cell proliferation,melanogenesis and antioxidant activity were determined.The related mechanism of inhibiting melanogenesis was also studied.The main contents and results are as follows:(1)In B16 melanoma cells,the effects of ginger essential oil on cell viability were determined by MTT assay,cell morphology observation,and flow cytometric assay.Results indicated that 20-100 ?g/mL of ginger essential oil significantly decreased the cell viability and the cells had unclear structure and retraction after treated with 100 ?g/mL of ginger essential oil for 48 h.Furthermore,the results of cell cycle indicated that cell cycle was arrested after treating with ginger essential oil for 24 h.The cells in G0/G1 and G2/M decreased,while cells in S phase increased.These results demonstrated that ginger essential oil could arrest the cell cycle at G2/M and S-G2 phase,which led to the inhibitory effects on cell proliferation.(2)The effects of ginger essential oil on melanogenesis and tyrosinase activity were studied.Results indicated that 20-100 ?g/mL of ginger essential oil could decrease intracellular melanin synthesis and melanin transportation;the results of tyrosinase activity showed that ginger essential oil inhibited intracellular tyrosinase activity,which demonstrated that ginger essential oil could decrease the melanin synthesis and affect melanin transportation by inhibiting the tyrosinase activity.(3)Intracellular glutathione level,superoxide dismutase activity,catalase activity,lipid peroxidation levels and reactive oxygen species were used as test indexes,and the effects of ginger essential oil on antioxidant activity were studied in B16 melanoma cells.Results showed that 20-100 ?g/mL of ginger essential oil improved the glutathione level,superoxide dismutase activity,and catalase activity.Furthermore,ginger essential oil inhibited the lipid peroxidation,production of intracellular reactive oxygen species,and extracellular reactive species.These results indicated that ginger essential oil had a certain antioxidant activity in melanoma cells,which was deduced to be one of mechanisms for ginger essential oil to inhibit the melanogenesis.(4)The effects of ginger essential oil on melanogenesis-related proteins were determined by Western Blotting.Results indicated that 20-80 ?g/ml of ginger essential oil decreased the expressions of tyrosinase(TYR),tyrosinase related protein-1(TRP1),tyrosinase related protein-2(TRP2),microphthalmia-associated transcription factors(MITF)and p38 mitogen-activated protein kinase pathway(p38 MAPK),which demonstrated that ginger essential oil decreased the intracellular melanin synthesis by inhibiting the expressions of tyrosinase related proteins,microphthalmia-associated transcription factors(MITF)and p38.Thus,the study demonstrated that ginger essential oil could inhibit the cell proliferation,and ginger essential oil suppressed the melanogenesis by controlled tyrosinase activity,expressions of related proteins and its antioxidant activity.It indicated that ginger essential oil could be used as a potent skin-whitening agent applied in food and cosmestics industry.