Objective In order to study the effects of Baiban granule-medicated serum on H2O2-induced oxidative stress(OS)and antioxidant gene expression in human epidermal melanocytes.Methods Normal epidermal melanocytes(Pigment cell 1,PIG1)were treated with proper concentration of H2O2 to establish OS model of melanocytes in vitro.Baiban granule herbal compound medicated serum was prepared.PIG1 at logarithmic growth stage were randomly divided into control group(PIG1+normal rat serum),model group(PIG1+H2O2),and Baiban granule group(PIG1+H2O2+Baiban granule medicated serum).Intracellular reactive oxygen species(ROS)were detected by flow cytometry.Malondialdehyde(MDA)was measured by thiobarbituric acid(TBA)assay.Real-time quantitative fluorescent polymerase chain reaction(RT-qPCR)was used to detect the mRNA expressions of nuclear factor erythroid-2-related factor 2(Nrf2),heme oxygenase 1(HO-1),glutathione peroxidase(GPx),superoxide dismutase 2(SOD2)and quinone oxidoreductase 1(NQO1).Results Compared with those in the control group,ROS and MDA levels in model group were significantly increased[ROS:(166.30±3.62)%vs.(100.00±3.20)%,P<0.01;MDA:(3.60±0.17)nmol/mg vs.(2.79±0.36)nmol/mg,P<0.01].Compared with model group,intracellular ROS and MDA levels in Baiban granule group were significantly decreased[ROS:(137.10±13.38)%vs.(166.30±3.62)%,P<0.05;MDA:(3.25±0.18)nmol/mg vs.(3.60±0.17)nmol/mg,P<0.01].Compared with the control group,GPx and SOD2 mRNA expressions in model group were significantly decreased(GPx mRNA:0.26±0.01 vs.1.00±0.12,P<0.01:SOD2 mRNA:0.25±0.02 vs.1.00±0.09,P<0.01),while Nrf2,HO-1 and NQO1 mRNA expressions were not statistically significant.Compared with the model group,GPx and SOD2 mRNA expressions in Baiban granule group were significantly increased(GPx mRNA:0.75±0.09 vs.0.26±0.01,P<0.01;SOD2 mRNA:0.62±0.10 vs.0.25±0.02,P<0.01),Nrf2 and HO-1 mRNA expressions were also significantly increased(Nrf2 mRNA:1.95±0.23 vs.1.11±0.34,P<0.05;HO-1 mRNA:1.91±0.21 vs.1.03±0.32,P<0.05),while the expression of NQO1 mRNA was not statistically significant.Compared with the control group,the expressions of Nrf2 and HO-1 mRNA in Baiban granule group were significantly increased(Nrf2 mRNA:1.95±0.23 vs.1.00±0.07,P<0.01;HO-1 mRNA:1.91±0.21 vs.1.00±0.23,P<0.01).Conclusion Baiban granule medicated serum can significantly reduce the levels of ROS and MDA in PIG1 under OS,which plays a protective role on oxidative damage.The mechanism may be related to activating nuclear factor erythroid-2-related factor2-antioxidant response element(Nrf2-ARE)signaling pathway and improving the mRNA expressions of the downstream antioxidant enzymes Gpx,SOD2 and HO-1.
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