The Study Of Effection And Possible Mechanism Of Stilbene Glycosides To Melanin Synthesis

Objectives As standards of living have risen and life style have change;vitiligo,chloasma hyperpigmentation more and more receive attention.Studies along with these natural medicines are getting more and more attention.Chinese medicine extracted from natural plants,are getting more and more concern because of little side effects.Polygonum multiflorum is one element of most chinese classical prescriptions which treat vitiligo,some scholars have found that the active ingredient of Polygonum multiflrum was stibene glucoside(THSG),and THSG can influence melanogenesis.culture melanocytes in three different density of THSG for 72 h,then detected the melanin synthesis and tyrosinase activity,cell cycle,ROS in the cell.analyze the effect of THSG on melanocytes in cultured environment.All the experiment were projected to supplement previous investigation,and to improve the treament of vitiligo.Provide more advice for chinicians and patients.Methods Normal human melanocytes were cultured in vitro,cells were randomly divided into five groups: the first group(0.1?g/ml),the second group(1?g/ml),the third group(12.5?g/ml),the fourth group(0?g/ml)group,the fifth group(?-MSH positive control group).Then,all the groups be cultured in the carbon dioxide incubator for 72 hours.Measured the melanocytes melanin syntheis by Na OH lysis method;and measure tyrosinase activity by DOPA oxidation.Distribution of cell cycle and ROS contents be examined by flow cytometry.At last,analyze the test results,and draw conclusions from the data.Results 1 Results of melanin content by NaOH lysis method show: After culture for 72 hours in the medium containing THSG,the melanin content is 166.66±0.10 when the concerntration of THSG is 0.1?g/ml;and the melanin content is 148.14±0.07 when the conterntion is 1?g/ml;when the concerntration is 12.5?g/ml,the result is 67.90±0.07.The result of the positive group is 148.14±0.07.Every group has significant effect compared with the blank control group(P<0.05).So the melanin content is increased when the concerntration is 0.1 and 1?g/ml,melanin content is reduced when the concerntration is12.5?g/ml.2 Results of tyrosinase activity by DOPA oxidation showed: After culture for72 hours in the medium containing THSG,the tyrosinase activity is 121.78±0.11 when the conterntion is 0.1?g/ml.and when the conterntion is 1 or 12.5?g/ml,the tyrosinase activity is 119.22±0.07 and 102.06±0.05;The result of the positive group is 245.35±0.77.The groups of 0.1 and 1?g/ml have significant effect compared with the blank control group(P<0.05),the tyrosinase activity increased.When the concerntion is12.5?g/ml,tyrosinase activity remained unchanged.3 Cell cycle: Cell cycle distribution of melanin detected by flow cytometry showed,after cultured in medium content MSH or THSG,the proportion of G0/G1 phase: 87.55±0.24%,85.20±0.18%,93.30±0.10%,and G0/G1 phase of blank control group is 92.52±0.52%.every group have significant effect compared with the blank control group(P<0.05).And the proportion of S phase:1.67±0.12%,1.78±0.04%,1.13±0.11%,S phase of blank control group is1.05±0.03%;1.67±0.12,1.78±0.04 have significant effect compared with the blank control group(P<0.05).4 ROS level: Ros level detected by flow cytometry showed,after cultured in the medium content THSG for 72 hours,ROS level of 0.1?g/ml group rised to121.33±0.27% of the control group,and 128.94±0.67% of 1?g/ml group.and112.95±0.45% of 12.5?g/ml group.Every group have significant effect compared with the blank control group(P<0.05).Conclusions 1 In certain concentrations(0.1,1?g/ml)THSG will promote norma l human melanocytes synthesis melanin and tyrosinase activity cultured in vitro.Can decrease number of S phase,increase ROS level.2 In certain concentrations(12.5?g/ml)THSG will inhibite normal human melanocytes synthesis melanin cultured in vitro.2 In certain concentrations,THSG will increase tyrosinase activity of Normal human melanocytes cultured in vitro.With the increase of concentration,tyrosinase activity were not affected.3 We found that low concentration of THSG promote melanocytes synthese,and high concentration of THSG inhibite melanin synthesis.THSG effect melanin synthesis not only through affect tyrosinase activity but also other access.4 This experiment provide further evidence for THSG on treating pigment disorder.