Adventitious Roots Extract Of Oplopanax Elatus Inhibits Melanin

Opolopanax elatus belongs to Araliaceae plants,which has precious medicinal value.However,due to its strict growing conditions and artificial over-excavation,the wild resources of Opolopanax elatus are destroyed,which affects the development and utilization of products.The cultured adventitious roots of Opolopanax elatus with bioreactors provide a large material to the product production.It has been proved that the adventitious roots of Opolopanax elatus have active ingredients such as flavonoids,phenols,and polysaccharides,and their anti-inflammatory,anti-oxidative and anti-cancer pharmacological activities have been preliminary studied,however there is no report on the inhibition of melanin.The plant composition has high safety and effects.Therefore,biochemical enzymology method and cytology method were used to study the effect of adventitious roots of Opolopanax elatus on the inhibition of melanin in order to provide a new plant material for the production of whitening cosmetics.Tyrosinase is the key enzyme for the formation of melanin in studies of melanin formation.Therefore in this study the biochemical enzymology method was used.Kojic acid was used as a control to optimize the enzyme activity conditions,and the final enzyme activity system was identified as the substrate dopa content was 1.0 mmol L-1,the tyrosinase content was 50 U mL-1,the constant temperature water bath time was 10 min,and the reaction time was 5 min;The ethanol extract showed the highest inhibitory effect on tyrosinase activity with IC50 was 0.39 mg mL-1.The effect of extract temperature,extract time,extract times and material-liquid ratio on tyrosinase activity was studied by using complete randomalized design.The extraction process was further optimized using orthogonal test methods.Through data analysis,the optimum extraction conditions were determined as follows:extract temperature was 70 ?,extract times was 3,extract time was 1.5 h,and solid-liquid ratio was 1:20,at this time,the inhibition rate of tyrosinase activity reached 77.6%.In cytology experiments,the effects of adventitious root extracts on melanin content in mouse melanoma cells(B16-F10)were studied,and the expression levels of MAPK and cAMP signaling pathway-related proteins were examined,in order to clarify the mechanism of the inhibition of melanin formation by adventitious root extracts.The results showed that the concentrations of the extracts from the adventitious roots ranged from 12.5 to 50 ?g mL-1,and the tyrosinase activity and the melanin content were significantly inhibited(p<0.01).In the MAPK signaling pathway,the p-p38(p<0.05)and p-JNK(p<0.01)protein expression were significantly decreased,and the p-ERK protein expression was significantly increased(p<0.01);in the cAMP signaling pathway related proteins CREB,the protein expression level of was significantly decreased(p<0.01)after treatment with the adventitious root extract.Further melanin formation-related transcription factors(MITF)and tyrosinase and its related proteins(TRP-1 and TRP-2)were studied and found that the expression of MITF and tyrosinase,TRP-1 and TRP-2 were significant(p<0.01)inhibitory effect.Therefore,adventitious root extracts can inhibit melanin by regulating the signaling pathways of MAPK and cAMP.