摘要
BACKGROUND: It has been suggested that melatonin (MT) can protect secondary neuronal injury. However, the protective effect of MT on neuronal injury in ischemia/reperfusion models in vitro still has not been proved. OBJECTIVE: To investigate the protective effect of MT on central ischemic injury of nerve cells and analyze its possible mechanism. DESIGN: Contrast observational study. SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Rats aged 7-8 days and weighing 10-12 g were provided by Medical Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technology. MT was provided by Sigma Company, USA. METHODS: The experiment was carried out in the Laboratory of Biochemistry and Molecular Biology, Tongji Hospital, Huazhong University of Science and Technology from October 2002 to March 2004. The effects of MT on the neurodegeneration induced by oxygen-glucose-deprivation (OGD) were tested in cultured rat cerebellar granule cells. Neuron damage was quantitatively assessed by Typan Blue exclusion and MTT assay at different time points after oxygen-glucose-deprivation (90 minutes). DNA gel electrophoresis and acridine orange stain were performed to determine the nature of cell damage. And fluorescence spectrophotometer was used for quantification of intracellular malondialdehyde (MDA) at various time intervals. MAIN OUTCOME MEASURES: Correlation between degrees of neuronal injury and reperfusion times, apoptosis, and production of MDA in cells. RESULTS: ① The neuron injury was aggravated with reperfusion time. ② The protective effect of MT was time- and dose-dependent when its concentration was not higher than 10 μmol/L. ③ When neurons were exposed to OGD for 90 minutes, part of the cells exhibited typical features of apoptosis: internucleosomal DNA condensation and DNA ladder on agarose gel electrophoresis. MT added to cells recovering from OGD exerted neuroprotective action against OGD-induced apoptosis. ④ In OGD exposed cultures, the production of MDA burst 12 hours after OGD, while MT significantly decreased the generation of MDA (P < 0.05) in a time-dependent manner. CONCLUSION: MT may have therapeutic potential in the prevention and treatment of ischemic/hypoxic neuronal damage, and this neuroprotective action may contribute to the antioxidant nature of MT.
BACKGROUND: It has been suggested that melatonin (MT) can protect secondary neuronal injury. However, the protective effect of MT on neuronal injury in ischemia/reperfusion models in vitro still has not been proved. OBJECTIVE: To investigate the protective effect of MT on central ischemic injury of nerve cells and analyze its possible mechanism. DESIGN: Contrast observational study. SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Rats aged 7-8 days and weighing 10-12 g were provided by Medical Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technology. MT was provided by Sigma Company, USA. METHODS: The experiment was carried out in the Laboratory of Biochemistry and Molecular Biology, Tongji Hospital, Huazhong University of Science and Technology from October 2002 to March 2004. The effects of MT on the neurodegeneration induced by oxygen-glucose-deprivation (OGD) were tested in cultured rat cerebellar granule cells. Neuron damage was quantitatively assessed by Typan Blue exclusion and MTT assay at different time points after oxygen-glucose-deprivation (90 minutes). DNA gel electrophoresis and acridine orange stain were performed to determine the nature of cell damage. And fluorescence spectrophotometer was used for quantification of intracellular malondialdehyde (MDA) at various time intervals. MAIN OUTCOME MEASURES: Correlation between degrees of neuronal injury and reperfusion times, apoptosis, and production of MDA in cells. RESULTS: ① The neuron injury was aggravated with reperfusion time. ② The protective effect of MT was time- and dose-dependent when its concentration was not higher than 10 μmol/L. ③ When neurons were exposed to OGD for 90 minutes, part of the cells exhibited typical features of apoptosis: internucleosomal DNA condensation and DNA ladder on agarose gel electrophoresis. MT added to cells recovering from OGD exerted neuroprotective action against OGD-induced apoptosis. ④ In OGD exposed cultures, the production of MDA burst 12 hours after OGD, while MT significantly decreased the generation of MDA (P < 0.05) in a time-dependent manner. CONCLUSION: MT may have therapeutic potential in the prevention and treatment of ischemic/hypoxic neuronal damage, and this neuroprotective action may contribute to the antioxidant nature of MT.
基金
the Natural Science Foundation of Hygienic Committee of Hubei Province, No: WJ01510
