Mailuoning suppresses H2O2-induced cortical neuronal injury and correlates with increased catalase activity and inhibited oxidative stress function

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 H202-induced primary cortical neuron injury in embryonic mice.DESIGN, TIME AND SETTING：Comparative observation and in v#ro 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; mitochondriai 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. Mai/uoning 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.