Regulation Of Tribulus Terrestris On Melanocyte Metabolism In SK-Mel-23 Cells

ObjectiveThis experiment explored the effects of different concentrations of Robinia pseudoacacia on melanin metabolism in human melanoma SK-Mel-23 cells,and preliminary explored the molecular mechanism of the regulation of pigment metabolism by Robinia pseudoacacia.MethodsIn this experiment,Tribulus terrestris extract as experimental drugs,the human melanoma SK-Mel-23 cells as a model,the experiment divided into Tribulus Terrestris group and H-89 + Tribulus terrestris group,MTT assay SK-Mel-23 Cell viability,melanin content was determined by NaOH cleavage method,tyrosinase activity was determined by oxidation rate method with dopa rate,and protein expression was detected by Western Blot method.Results1.Tribulus terrestris group of melanin metabolism and mechanism of action1.1 Effect on cell activity The cell viability of SK-Mel-23 cells treated with different concentrations of Tribulus terrestris was statistically significant at 24,48 and 72 hours(F=19.623,P<0.001;F=5.49,P<0.01;F=2.04,P<0.01).At low concentration,the activity of the cells showed a promoting effect on the cell activity,with inhibition as the concentration increased.1.2 Effect on melanin synthesis The concentrations of Tribulus terrestris in SK-Mel-23 cells after 24 hours,48 hours and 72 hours,were all statistically significant for melanin synthesis(F = 11.927,P<0.001;F = 9.03,P<0.001;F ?16.5,P<0.001),inhibited the melanogenesis of SK-Mel-23 cells at low concentration,and promoted the increase of the concentration of melanin.1.3 Effect on tyrosinase activity The tyrosinase activities of SK-Mel-23 cells after 24 hours,48 hours and 72 hours of different concentrations of Tribulus terrestris were statistically significant(F = 5.74,P<0.001;F = 3.97,P<0.001;F =21.55,P<0.001).The SK-Mel-23 cell tyrosinase activity was inhibited at low concentration,which showed a positive effect with increasing concentration.1.4 Effect on protein expression The effects of Tribulus terrestris on the expression of TYR,TRP-1,MITF and p-CREB in SK-Mel-23 cells after 48h were statistically significant(F = 23.97,P<0.001;F = 11.37,P<0.001;F = 1.53,P<0.05;F = 2.67,P<0.05;),the protein expression was inhibited at low concentration(1.02mg/ml-2.05mg/ml)to promote protein expression.2.Effects of H-89 + Tribulus terrestris group on melanin metabolism and protein expression2.1 Effect on melanin synthesis H-89 treatment 1 hour after adding different concentrations of Tribulus terrestris culture 48h,different concentrations of the group and the control group added H-89 compared to melanin synthesis had no significant difference(F = 0.683,P = 0.61),different concentrations and The effect of adding H-89 control group to melanin synthesis was statistically significant(F =2915,P<0.001).2.2 Effect on tyrosinase activity H-89 treatment 1 hour after adding different concentrations of Tribulus terrestris cultured 48h,different concentrations of H-89 control group and compared to the ratio of tyrosinase activity was not statistically significant(F=0.416,P=0.822),different concentrations There was no significant difference in the activity of tyrosinase compared with the control without H-89(F=354.9,P<0.001).2.3 Effect on protein expression H-89 treatment for 1 hour and then added different concentrations of Tribulus terrestris for 48h.There was no significant difference in the expression of TYR,TRP-1,MITF and p-CREB between H-89 and control group(F=2.499,P=0.9;F=3.062,P=0.89;F=0.76,P=0.61;F=0.89,P=0.51).The protein expression did not change with the concentration.Protein expression was statistically significant(F=7.324,P<0.05;F=2.927.P<0.05;F=10.41,P<0.01;F=7.233,P<0.001).ConclusionThrough the study of this topic found that Tribulus terrestris biomembrane regulation of biometrical existence,the best experiment time is 48h.Tribulus terrestris inhibited the cell viability at low concentration,promoted the cell viability at high concentration,inhibited the synthesis of melanin and tyrosinase activity at low concentration,and promoted the activity at high concentration,The mechanism may be the inhibition of the expression of TYR,TRP-1,MITF and p-CREB protein at low concentration,and the expression of TYR,TRP-1,MITF and p-CREB protein at high concentration.After addition of PKA pathway inhibitor,different concentrations of Tribulus terrestris did not change the pigment metabolism,that is,Tribulus terrestris regulated the pigment metabolism by regulating the cAMP/PKA pathway.