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14 April 2016 : Laboratory Research  

Protective Effect of Puerarin Against Oxidative Stress Injury of Neural Cells and Related Mechanisms

Yuan ChengABCD, Wei LengDE, Jingshu ZhangFG

DOI: 10.12659/MSM.896058

Med Sci Monit 2016; 22:1244-1249

Abstract

BACKGROUND: Parkinson’s disease (PD) is manifested as degeneration of dopaminergic neurons in substantia nigra compacta. The mitochondrial dysfunction induced by oxidative stress is believed to a major cause of PD. Puerarin has been widely applied due to its estrogen nature and anti-oxidative function. This study thus investigated the protective role of puerarin against oxidative stress injury on PC12 neural cells, in addition to related mechanisms.

MATERIAL AND METHODS: PC12 cells were pre-treated with gradient concentrations of puerarin, followed by the induction of 0.5 mM H2O2. MTT assay was used to detect cell viability. Enzyme-linked immunosorbent assay (ELISA) was employed to detect intracellular level of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH). Cell apoptosis was determined by Annexin-V/7-AAD double labelling. Reactive oxidative species (ROS) and lactate dehydrogenase (LDH) activities were then measured. Cellular levels of caspase-3 and caspase-9 were also determined.

RESULTS: The pre-treatment using puerarin significantly reversed H2O2-induced oxidative stress injury, as it can increase proliferation, SOD and GSH activities, decrease MDA activity, suppress apoptosis of PC12 cells, and decrease ROS and LDH production (p<0.05 in all cases). Further assays showed depressed up-regulation of caspase-3 and caspase-9 after puerarin pretreatment.

CONCLUSIONS: Puerarin pretreatment can decrease activity of caspase-3 and caspase-9 activity in PC12 cells, thus protecting cells from oxidative injury.

Keywords: Antioxidants - pharmacology, Caspase 9 - metabolism, Cell Survival - drug effects, Glutathione - metabolism, Isoflavones - pharmacology, Malondialdehyde - metabolism, Neurons - pathology, Neuroprotective Agents - pharmacology, Oxidative Stress - drug effects, Reactive Oxygen Species - metabolism, Superoxide Dismutase - metabolism

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750