论文部分内容阅读
BACKGROUND:Mailuoning,a Chinese herb,has been widely used in China to treat acute ischemic stroke,and the major component exhibits anti-oxidative effects.However,the precise anti-oxidation pathway remains uncertain.OBJECTIVE:To validate the protective effects of Mailuoning on H2O2-induced primary cortical neuron injury in embryonic mice.DESIGN,TIME AND SETTING:Comparative observation and in vitro experiments were performed at the Jiangsu Key Laboratory for Molecular Medicine from January 2008 to September 2009.MATERIALS:Mailuoning(Nanjing Jinling Medical Company,China),reactive oxygen species(ROS) kit(Beyotime Biotechnology,China),superoxide dismutase(SOD),Cu/Zn SOD kit,malondialdehyde(MDA) kits(Nanjing Jiancheng,China),mitochondrial membrane potential(GMS10013.1,GENMED,USA) and catalase activity assay kit(Beyotime Biotechnology,China) were utilized for the present study.METHODS:Mouse embryonic cortical neurons were isolated and cultured with culture medium containing H2O2(80 μmol/L) and/or Mailuoning(1.25 μg/mL) for 24 hours.MAIN OUTCOME MEASURES:Neuronal viability and death were detected by methyl thiazolyl tetrazdium and flow cytometry;ROS production was determined by flow cytometry;mitochondrial membrane potential was detected using fluorescent staining;SOD activity was detected using a modified nitroblue tetrazolium method;Cu/Zn SOD and catalase activity was detected by spectrophotometry;and MDA was determined using the lipid peroxidation method.RESULTS:H2O2 increased ROS production and MDA concentration(P < 0.05),and decreased mitochondrial membrane potential,SOD,Cu/Zn SOD and catalase activity(P < 0.05);the number of surviving neurons(P < 0.05) was also reduced.Mailuoning reversed these changes.CONCLUSION:Mailuoning protects H2O2-induced injury in cortical cells by inhibiting ROS and MDA,increasing depolarization of mitochondrial membrane,and enhancing SOD and catalase activity.
BACKGROUND: Mailuoning, a Chinese herb, has been widely used in China to treat acute ischemic stroke, and the major component exhibits anti-oxidative effects. However, the precise anti-oxidation pathway remains uncertain. OBJECTIVE: To validate the protective effects of Mailuoning on H2O2-induced primary cortical neuron injury in embryonic mice. DIGNIGN, TIME AND SETTING: Comparative observation and in vitro experiments were performed at the Jiangsu Key Laboratory for Molecular Medicine from January 2008 to September 2009. MAIALIALS: Mailuoning (Nanjing Jinling Medical Company, (Reactive oxygen species (ROS) kit (Beyotime Biotechnology, China), superoxide dismutase (SOD), Cu / Zn SOD kit, malondialdehyde (MDA) kits (Nanjing Jiancheng, China), mitochondrial membrane potential (GMS10013.1, GENMED , USA) and catalase activity assay kit (Beyotime Biotechnology, China) were utilized for the present study. METHODS: Mouse embryonic cortical neurons were isolated and cultured with culture medium containing H2O2 (80 μmol / L) and / or Mailuoning (1.25 μg / mL) for 24 hours. MAIN OUTCOME MEASURES: Neuronal viability and death were detected by methyl thiazolyl tetrazdium and flow cytometry; ROS production was determined by flow cytometry; mitochondrial membrane potential was detected using fluorescent staining; SOD activity was detected using a modified nitroblue tetrazolium method; Cu / Zn SOD and catalase activity was detected by spectrophotometry; and MDA was determined using the lipid peroxidation method .RESULTS: H2O2 increased ROS production and MDA concentration (P <0.05), and decreased mitochondrial The number of surviving neurons (P <0.05) was also reduced. Mailu-t switched these changes. CONCLUSION: Mailuoning protects H2O2-induced injury in cortical cells by inhibiting ROS and MDA, increasing depolarization of mitochondrial membrane, and enhancing SOD and catalase activity.