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BACKGROUND: Evidence illustrates that androgen has a neuroprotective role. However, whether androgen also has the protective effect on hippocampal neurons during free radical mediated injury remains unclear.OBJECTIVE: To investigate the neuroprotective effect of androgen on hippocampal neurons during free radical damage.DESIGN, TIME AND SETTING: A controlled in vitro experiment was performed at the Department of Human Anatomy, Cell Culture Lab, and Neuroendocrinology Lab, Basic Medical School, Hebei Medical University from February to June 2009.MATERIALS: Testosterone was provided by Tianjin Jinyao Amino Acid Company, China.METHODS: Primary cultured neurons from 24 Sprague Dawley rats were randomly assigned into four groups: control, H_2O_2, testosterone, and testosterone (pre-added) plus H_2O_2 groups.MAIN OUTCOME MEASURES: The positive cell ratio of microtubule associated protein-II and neuron specific enolase was determined by immunocytochemistry. Neuronal morphology was observed by hematoxylin-eosin staining and Nissl staining. Cell vitality and viability were determined using an inverted phase contrast microscope. The content of nitric oxide synthase, malondialdehyde, and superoxide dismutase were measured with a spectrophotometer.RESULTS: As compared with the control group, cell vitality and viability, and superoxide dismutase level were significantly decreased in the H_2O_2 group (P < 0.05), while nitric oxide synthase and malondialdehyde levels were significantly increased (P < 0.05). Neuronal vitality and viability as well as superoxide dismutase level in the testosterone plus H_2O_2 group were significantly greater than in the H_2O_2 group (P < 0.05), and nitric oxide synthase and malondialdehyde levels were significantly less than in the H_2O_2 group (P < 0.05).CONCLUSION: Androgen partially reversed H_2O_2-induced neuronal damage and protected neurons.