白斑冲剂含药血清对H_(2)O_(2)诱导的正常人表皮黑色素细胞氧化应激和抗氧化基因表达的影响  

Effect of Baiban Granule-medicated Serum on H_(2)O_(2)-induced Oxidative Stress and Antioxidant Gene Expression in Human Epidermal Melanocytes

作　　者：韩静倩 姚卫锋[1] 李欢[1] 李芸芸[1] 赵艳霞[1] Han Jingqian;Yao Weifeng;Li Huan;Li Yunyun;Zhao Yanxia(Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital,Tianjin 300120,China)

机构地区：[1]天津市中医药研究院附属医院,天津300120

出　　处：《中国中西医结合皮肤性病学杂志》2024年第4期289-294,299,共7页Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine

基　　金：2021年天津市卫生健康委员会中医;中西医结合科研项目(编号:2021085)。

摘　　要：目的 研究白斑冲剂含药血清对过氧化氢(H_(2)O_(2))诱导的正常人表皮黑色素细胞(PIG1)氧化应激(OS)和抗氧化基因表达的影响。方法 以适当浓度H_(2)O_(2)处理PIG1,建立体外黑色素细胞OS模型。配制白斑冲剂中药复方含药血清。将对数生长期的PIG1随机分为对照组(PIG1+正常大鼠血清)、模型组(PIG1+H_(2)O_(2))和白斑冲剂组(PIG1+白斑冲剂含药血清+H_(2)O_(2))。流式细胞仪检测各组细胞内活性氧(ROS);硫代巴比妥酸(TBA)法检测各组细胞内丙二醛(MDA);实时荧光定量聚合酶链反应(RT-qPCR)检测抗氧化基因核因子E2相关转录因子2(Nrf2)、血红素氧合酶1(HO-1)、谷胱甘肽过氧化物酶(Gpx)、超氧化物歧化酶2(SOD2)和醌氧化还原酶1(NQO1)mRNA的表达。结果 与对照组比较,模型组细胞内ROS和MDA水平显著升高[ROS:(166.30±3.62)%比(100.00±3.20)%,P<0.01;MDA:(3.60±0.17) nmol/mg比(2.79±0.36) nmol/mg,P<0.01];与模型组比较,白斑冲剂组细胞内ROS和MDA水平显著下降[ROS:(137.10±13.38)%比(166.30±3.62)%,(P<0.05);MDA:(3.25±0.18) nmol/mg比(3.60±0.17) nmol/mg,P<0.01)];与对照组比较,模型组细胞中GPx和SOD2 mRNA的表达显著下降(GPx mRNA:0.26±0.01比1.00±0.12,P<0.01;SOD2 mRNA:0.25±0.02比1.00±0.09,P<0.01),Nrf2、HO-1和NQO1 mRNA的表达差异无统计学意义(P>0.05);与模型组比较,白斑冲剂组细胞中GPx和SOD2 mRNA的表达显著升高(GPx mRNA:0.75±0.09比0.26±0.01,P<0.01;SOD2mRNA:0.62±0.10比0.25±0.02,P<0.01),Nrf2和HO-1 mRNA的表达也明显升高(Nrf2 mRNA:1.95±0.23比1.11±0.34,P<0.05;HO-1 mRNA:1.91±0.21比1.03±0.32,P<0.05),NQO1 mRNA的表达差异无统计学意义(P>0.05);与对照组比较,白斑冲剂组Nrf2和HO-1 mRNA的表达明显升高(Nrf2 mRNA:1.95±0.23比1.00±0.07,P<0.01;HO-1 mRNA:1.91±0.21比1.00±0.23,P<0.01)]。结论 白斑冲剂含药血清能明显降低OS下PIG1内ROS和MDA的含量,有抗氧化损伤的作用。其作用机制可能是与激活Nrf2-ARE信号通路,提高下游抗氧�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 H_(2)O_(2) 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+H_(2)O_(2)),and Baiban granule group(PIG1+H_(2)O_(2)+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(SOD_(2))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.

关 键 词：白癜风 白斑冲剂 氧化应激 黑色素细胞 活性氧簇 核因子E2相关因子-抗氧化反应元件信号通路 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]