Effects Of Yunnan Camellia And Several Plants Extracts On The Melanin Synthesis And Tyrosinase Activity Of Human Epidermal Melanocytes

Objective:Screen out the effective ingredients and concentrations from extracts of Yunnan camellia, Chloranthus japonicas, Dodonaea viscose, Paris polyphylla, which have security and skin whitening function, as to provide theoretical basis for the development of plant whitening agent.Methods:Thawing human epidermal melanocytes (light), maintenance the culture and subculture the cell, seeding the3-10generation cells which in logarithmic growth phase in96-well culture plate. Set up7different concentration gradient (10μg/mL、25μg/mL、50μg/mL、100μg/mL、200μg/mL、400μg/mL、800μg/mL) for this7different kinds of plants extracts (Yunnan camellia leaves extract,70%ethanol elution of Yunnan camellia leaves extract, Yunnan camellia flower bud extract,70%ethanol elution of Yunnan camellia bud extract, Chloranthus japonicas, Dodonaea viscose, Paris polyphylla), work on the human epidermal melanocytes culture in vitro for72h respectively, each different concentration drug for three repeated hole, put arbutin as the positive control (HEM-m+MelM+Arbutin100μg/mL,100uL/hole), set the negative control(HEM-m+MelM) and the blank control(MelM), set up3repeated hole for each group. Using MTT to measure the influence of different ingredients and concentrations on the HEM-m proliferation activity, and measure the effect on tyrosinase activity through L-Dopa oxidation reaction in vitro, using NaOH-lysis to test melanin synthesis of HEM-m. Each experiment repeated for3times.Results:1the influence on HEM-m proliferation activity:compare to the negative control,70%ethanol elution of Yunnan camellia leaves extract and Dodonaea viscose had no obvious effect on HEM-m proliferation at10μg/mL (P>0.05), the rest different concentration drugs and arbutin all showed inhibiting the proliferation activity of HEM-m obviously (P<0.05or P<0.01); Compare to the positive control(arbutin), when the Yunnan camellia leaves extract at400μg/mL,70%ethanol elution of Yunnan camellia leaves extract at100-10μg/mL, Yunnan camellia flower bud extract at25-10μg/mL,70%ethanol elution of Yunnan camellia bud extract at10μg/mL, Dodonaea viscose at25-10μg/mL, the proliferation activity of HEM-m was same as arbutin (P>0.05). the rest drugs compare with positive control, is statistically different,(P<0.05or P<0.01), with Yunnan camellia leaves extract had the stronger suppressive effect on HEM-m proliferation at800μg/mL, and the rest of the drugs showed a inferior suppressive effect.2the influence on HEM-m tyrosinase activity:compare to the negative control, the tyrosinase activity of arbutin and all plants extract at any concentration was obviously lower than the negative control (P<0.05or P<0.01); Compare with the positive control, when Yunnan camellia leaves extract at10μg/mL,70%ethanol elution of Yunnan camellia leaves extract at400、200、100、50、25、10μg/mL, Yunnan camellia flower bud extract at25、10μg/mL,70%ethanol elution of Yunnan camellia bud extract at100、50μg/mL, Chloranthus japonicas at400、200、100、50、25、10μg/mL, Dodonaea viscose at any concentration, Paris polyphylla at200、100、50、25、10μg/mL, the tyrosinase activity had no obvious different (P>0.05). The tyrosinase activity of the rest drugs compare with positive control, is statistically different,(P<0.05or P<0.01), with the70%ethanol elution of Yunnan camellia bud extract at25、10μg/mL had the higher activity and the rest drugs had the lower activity than arbutin.Conclusions:the security and skin whitening effect of Yunnan camellia leaves extract at800μg/mL was superior than arbutin, when Yunnan camellia leaves extract at400μg/mL, the effect of skin whitening was superior than arbutin although the security was same as it. When70%ethanol elution of Yunnan camellia leaves extract at100-10μg/mL, Yunnan camellia flower bud extract at25-10μg/mL, Dodonaea viscose at25-10μg/mL, the security and effect of skin whitening were same as arbutin. The concentration of those plant extracts can be used as a new plant skin whitening agent to research and develope in the future.