Effects Of Imperatorin, Isoimpemtorin, Paeoniflorin On Biological Activity And Migration Of Human Epidermal Melanocytes

Objective: Through establishing melanocyte (MC) culture and stimulateing it withtwo monomers of radix angelicae (imperatorin, isoimpemtorin) and a monomer ofradix paeoniae alba (paeoniflorin)respectively. We observed the effects of variesmonomers on biological activities and migration of human epidermal melanocytes.It helps us to discuss the possible mechanism of three kinds of Chinese medicinemonomer on the melanocytes and provide new dieas for the treatment of pigmentdisorders.Methods:1.We established a culture system which is a simulation ofphysiological state of melanocytes.2.We detected the effects of Chinese herbsmonomers on proliferation activity of human epidermal melanocytes by MTTmethod respectively.3.We explored the effects of monomers on tyrosinase activityof human epidermal melanocytes by L-DOPA method respectively.4.We detectedthe effects of monomers on melanogenesis of human epidermal melanocytes byNaOH method respectively.5.We used transwell assay to explore the effects ofmonomers on migration of human epidermal melanocytes respectively. All thedata were analyzed by SPSS17.0statistics software.Results:1.The20μg/mL,50μg/mL imperatorin group，the5μg/mL,10μg/mL and20μg/mL isoimpemtorin group，the5μg/mL,10μg/mL,20μg/mL and50μg/mL paeoniflorin group，all showed the role of promoting melanocyte proliferation.When groups of monomers at the concentraton of100μg/mL,there were notobvious inhibitory effects on cell proliferation, the melanocyte still grew well.Groups of monomers’effects on promoting melanocytes proliferation hadconcentration-dependent characteristics when the concentrations were low, butwith high concentrations the ability of promoting melanocytes proliferationdeclined instead. According to the experimental results,10μg/mL was the besteffect concentration of the paeoniflorin group and the psoralen group; so was20μg/mL to the imperatorin group and the isoimpemtorin group. And the5μg/mL,10μg/mL and20μg/mL isoimpemtorin group, their proliferation abilitieswere stronger than the psoralen group with the same concentration.2. The20μg/mL,50μg/mL imperatorin group, the10μg/mL,20μg/mL isoimpemtoringroup，the10μg/mL,20μg/mL,50μg/mL paeoniflorin group, they all showed thethe role of increasing tyrosinase activity of melanocytes. And their efficacies weresimilar to the psoralen group. Groups of monomers’ effects on increasingmelanocytes tyrosinase activity also had concentration-dependent characteristicswhen the concentrations were low, but with high concentrations the ability ofincreasing melanocytes tyrosinase activity declined instead. And10μg/mL was thebest effect concentration of the isoimpemtorin group and the paeoniflorin group;so was20μg/mL to the imperatorin group and the psoralen group.3. Theimperatorin group did not showed the ability of promoting the synthesis ofmelanin at the concentration of2.5μg/mL,5μg/mL,10μg/mL,20μg/mL,50μg/mL, 100μg/mL. The20μg/mL isoimpemtorin group,the20μg/mL,50μg/mLpaeoniflorin group,they all promoted the synthesis of melanin.The isoimpemtoringroup,the paeoniflorin group and the psoralen group,their abilities of promotingthe synthesis of melanin all had concentration-dependent characteristics when theconcentrations were low, but with high concentrations the ability of promoting thesynthesis of melanin declined instead. And20μg/mL was the best effectconcentration of the isoimpemtorin group and the psoralen group;so was50μg/mLto the paeoniflorin group.4.The imperatorin group did not showed the ability ofpromoting migration of melanocyte at the concentrations of10μg/mL,20μg/mL,50μg/mL. The20μg/mL and50μg/mL isoimpemtorin group promotedthe migration of melanocytes and there was significant difference statistically(P<0.01)， and the ability of promoting the migration of melanocytes hadconcentration-dependent characteristics. The20μg/mL paeoniflorin grouppromoted the migration of melanocytes and there was significant differencestatistically (P<0.01).And the psoralen group promoted the migration ofmelanocytes at the concentrations of10μg/mL,20μg/mL,50μg/mL,in which20μg/mL was the best concentration for the migration. The isoimpemtorin groupand the psoralen group at the concentration of50μg/mL all promoted themigration of melanocyte and there was significant difference statistically(P<0.01),and their strength was equal.Conclusion:1.The imperatorin,isoimpemtorin and paeoniflorin all have the abilityof promoting the melanocytes proliferation, and when groups of monomers at high concentration of100μg/mL, there are not obvious inhibitory effects on cellproliferation, it confirms the safety of three monomers. And the effects onpromoting the melanocytes proliferation of isoimpemtorin group is better than thepsoralen group.2. The imperatorin, isoimpemtorin and paeoniflorin all have theabilities of increasing tyrosinase activity,and their efficacies are similar to thepsoralen group.3.The isoimpemtorin and paeoniflorin all have the abilities ofpromoting the synthesis of melanin,whereas the imperatorin was not detected hasthe ability.4.The20μg/mL,50μg/mL isoimpemtorin group and the20μg/mLpaeoniflorin group promote the migration of melanocytes; whereas the imperatoringroup does’ nt show the ability of promoting the migration of melanocytes at theconcentration of10μg/mL,20μg/mL,50μg/mL. Above all, we speculate that theisoimpemtorin is the major monomer of radix angelicae, the paeoniflorin is themajor monomer of radix paeoniae alba.The50μg/mL isoimpemtorin group and50μg/mL psoralen group have the equal abilities of promoting the migration ofmelanocytes.