OBJECTIVE To investigate the neuroprotective effects of hesperetin on central neurons under chronic high glucose,and the relationship to glyoxalase 1(Glo-1),a cytoprotective enzyme.METHODS The human neuroblas⁃toma SH-...OBJECTIVE To investigate the neuroprotective effects of hesperetin on central neurons under chronic high glucose,and the relationship to glyoxalase 1(Glo-1),a cytoprotective enzyme.METHODS The human neuroblas⁃toma SH-SY5Y cells were divided into 5 groups:normal glucose,high glucose(HG),HG plus low,middle,or high concentra⁃tion of hesperetin(1,5,25μmol·L^-1).After treatment for 72 h,neuron damages,Glo-1 expressions and functions,as well as Nrf2/ARE pathway and its regulating mechanisms were examined.RESULTS Hesperetin increased cell viability and decreased lactate dehydrogenase release,which was accompanied by the elevated activity,protein,and mRNA levels of Glo-1 as well as the enhanced Glo-1 functions in SH-SY5Y cells cultured with HG.Moreover,hesperetin activated Nrf2/ARE pathway as evidenced by the raised Nrf2 and p-Nrf2 levels in nucleus and up-regulation of γ-glutamycysteine synthase(γ-GCS),a well-known target gene of Nrf2/ARE pathway.Nevertheless,pretreatment with a PKC inhibitor(Go 6983)or an Akt inhibitor(MK-22062HCl,reflecting GSK-3β activation)abolished the effect of hesperetin on protein expressions of Glo-1 and γ-GCS.CONCLUSION Hesperetin exerted the neuroprotection by promoting Glo-1 function in central neurons in long-term HG condition,which was mediated by activation of Nrf2/ARE pathway;moreover,the increased Nrf2 phosphorylation and nuclear translocation mediated by PKC activation and/or GSK-3β inhibition were involved in the activation of Nrf2/ARE pathway by hesperetin.展开更多
建立了同时测定加味四逆散中柴胡皂苷a、芍药苷、橙皮苷、橙皮素、甘草酸和甘草苷的超高效液相色谱-质谱(UPLC-MS)方法,并应用于复方的药代动力学研究。色谱柱为Agilent Zorbax Eclipse Plus C_(18)柱(100×2.1mm,1.8μm);流动相为0...建立了同时测定加味四逆散中柴胡皂苷a、芍药苷、橙皮苷、橙皮素、甘草酸和甘草苷的超高效液相色谱-质谱(UPLC-MS)方法,并应用于复方的药代动力学研究。色谱柱为Agilent Zorbax Eclipse Plus C_(18)柱(100×2.1mm,1.8μm);流动相为0.1%甲酸溶液-乙腈;体积流量0.5mL/min;进样量为5μL;采用电喷雾离子源负离子模式(ESI-),多反应监测(MRM)模式进行测定。方法对6种化合物测定在一定的浓度范围具有良好的线性关系(r^2≥0.998),日内和日间精密度(RSD)值均小于8.3%,准确度为94.3%~104.9%,基质效应在82.5%~101.6%合理范围内,提取回收率在85.8%~94.3%之间。经方法学考察,该方法快速、成本低,可同时监测加味四逆散中6种生物活性成分的血药浓度。展开更多
基金National Natural Science Foundation of China(81371210)
文摘OBJECTIVE To investigate the neuroprotective effects of hesperetin on central neurons under chronic high glucose,and the relationship to glyoxalase 1(Glo-1),a cytoprotective enzyme.METHODS The human neuroblas⁃toma SH-SY5Y cells were divided into 5 groups:normal glucose,high glucose(HG),HG plus low,middle,or high concentra⁃tion of hesperetin(1,5,25μmol·L^-1).After treatment for 72 h,neuron damages,Glo-1 expressions and functions,as well as Nrf2/ARE pathway and its regulating mechanisms were examined.RESULTS Hesperetin increased cell viability and decreased lactate dehydrogenase release,which was accompanied by the elevated activity,protein,and mRNA levels of Glo-1 as well as the enhanced Glo-1 functions in SH-SY5Y cells cultured with HG.Moreover,hesperetin activated Nrf2/ARE pathway as evidenced by the raised Nrf2 and p-Nrf2 levels in nucleus and up-regulation of γ-glutamycysteine synthase(γ-GCS),a well-known target gene of Nrf2/ARE pathway.Nevertheless,pretreatment with a PKC inhibitor(Go 6983)or an Akt inhibitor(MK-22062HCl,reflecting GSK-3β activation)abolished the effect of hesperetin on protein expressions of Glo-1 and γ-GCS.CONCLUSION Hesperetin exerted the neuroprotection by promoting Glo-1 function in central neurons in long-term HG condition,which was mediated by activation of Nrf2/ARE pathway;moreover,the increased Nrf2 phosphorylation and nuclear translocation mediated by PKC activation and/or GSK-3β inhibition were involved in the activation of Nrf2/ARE pathway by hesperetin.
文摘建立了同时测定加味四逆散中柴胡皂苷a、芍药苷、橙皮苷、橙皮素、甘草酸和甘草苷的超高效液相色谱-质谱(UPLC-MS)方法,并应用于复方的药代动力学研究。色谱柱为Agilent Zorbax Eclipse Plus C_(18)柱(100×2.1mm,1.8μm);流动相为0.1%甲酸溶液-乙腈;体积流量0.5mL/min;进样量为5μL;采用电喷雾离子源负离子模式(ESI-),多反应监测(MRM)模式进行测定。方法对6种化合物测定在一定的浓度范围具有良好的线性关系(r^2≥0.998),日内和日间精密度(RSD)值均小于8.3%,准确度为94.3%~104.9%,基质效应在82.5%~101.6%合理范围内,提取回收率在85.8%~94.3%之间。经方法学考察,该方法快速、成本低,可同时监测加味四逆散中6种生物活性成分的血药浓度。
