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. OBJECTIV...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, H202, testosterone, and testosterone (pre-added) plus H2O2 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 H202 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 H2O2 group were significantly greater than in the H2O2 group (P 〈 0.05), and nitric oxide synthase and malondialdehyde levels were significantly less than in the H2O2 group (P〈 0.05). CONCLUSION: Androgen partially reversed H2O2-induced neuronal damage and protected neurons.展开更多
The effects of bis(7) tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate induced cell injury were investigated in primary cerebral cortical neurons of rats. Exposure of cultured neurons (1...The effects of bis(7) tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate induced cell injury were investigated in primary cerebral cortical neurons of rats. Exposure of cultured neurons (12 days after plating) to 0.5 mmol/L glutamate for 30 min resulted in significant cell damage. Pretreatment with bis(7) tacrine (0.03 1.0 μmol/L) reduced the glutamate induced neurotoxicity in a concentration dependent manner and the maximal response was seen at 1 μmol/L with approximately 30% protection. A receptor binding assay showed that bis(7) tacrine can completely displace MK 801 binding to rat cortical membrane with an IC 50 of 0.57 μmol/L. These findings suggest that bis(7) tacrine can directly interact with N methyl D aspartate receptor channel complex, which may contribute to the inhibitor's protective effects against glutamate induced excitotoxicity. Thus, it is possible that anti glutamate/anti AChE synergism is responsible for potentially better Alzheimer's therapy of bis(7) tacrine relative to tacrine.展开更多
Primary cultures of rat cerebral corticai neurons were prepared from conices or 2 days Sprague-Dawlley rats.We made an attempt to imitate the hypoxic neurons injury for testing neuroprotective erfect or Sibelium. It ...Primary cultures of rat cerebral corticai neurons were prepared from conices or 2 days Sprague-Dawlley rats.We made an attempt to imitate the hypoxic neurons injury for testing neuroprotective erfect or Sibelium. It was found that Sibelium could significantly decrease the amounts of LDH leaking from intracellular fluid and increase the neuron-membrane fluidity of hypoxic neurons.Hypoxic group had significantly higher incorporation of 3H-TDR and 3H-UR than control group,while Sibelium could decrease the incorporation of 3H-TDR and 3H-UR. In addition,Sibelium could decrease the incorporation of 3H-TDR and 3H-UR in normal cultured neurons.The results showed that Sibelium could protect the neuronmembranes against alteration caused by hypoxia and regulate the synthesis of DNA and RNA.展开更多
The prevalence of domestic and industrial electrical appliances has raised concerns about the health risk of extremely low-frequency magnetic fields(ELF-MFs). At present, the effects of ELF-MFs on the central nervou...The prevalence of domestic and industrial electrical appliances has raised concerns about the health risk of extremely low-frequency magnetic fields(ELF-MFs). At present, the effects of ELF-MFs on the central nervous system are still highly controversial, and few studies have investigated its effects on cultured neurons. Here, we evaluated the biological effects of different patterns of ELF-MF exposure on primary cultured hippocampal neurons in terms of viability, apoptosis, genomic instability,and oxidative stress. The results showed that repeated exposure to 50-Hz 2-mT ELF-MF for 8 h per day after different times in culture decreased the viability and increased the production of intracellular reactive oxidative species in hippocampal neurons. The mechanism was potentially related to the up-regulation of Nox2 expression.Moreover, none of the repeated exposure patterns had significant effects on DNA damage, apoptosis, or autophagy, which suggested that ELF-MF exposure has no severe biological consequences in cultured hippocampal neurons.展开更多
Objective Muncl8-1 has an important role in neurotransmitter release, and controls every step in the exocy- totic pathway in the central nervous system. In the present study, whether epileptic seizure causes a change ...Objective Muncl8-1 has an important role in neurotransmitter release, and controls every step in the exocy- totic pathway in the central nervous system. In the present study, whether epileptic seizure causes a change of Muncl8 localization in neuronal nuclei was analyzed. Methods Epilepsy models were established by injection of kainic acid (KA) solution into hippocampus of Sprague-Dawley (SD) rats or intraperitoneal injection of KA in Kunming mice. The hippocampal neurons were prepared from embryonic day 18 SD rats, and cultured in neurobasal medium, followed by treatment with glutamate for 3 h. Neuronal and glial nuclei of hippocampus were separated by sucrose density gradient centrifugation. The nucleus-enriched fractions were stained with 0.1% Cresyl Violet for morphological assay. Immuno- chemistry and immunoelectron microscopy with anti-Muncl 8-1 antibody were used to determine the nuclear locatization of Munc 18-1. Immunoblotting was used to detect the protein level of Munc 18-1. Results The localization of Munc 18-1 in nucleus of rat hippocampal neuron was confirmed by immunochemistry, immunoelectron microscopy, and immunob- lotting detection of neuronal nucleus fraction. In animals receiving intrahippocampal or intraperitoneal injection of KA, immunostaining revealed that the expression of Muncl 8-1 decreased in pyramidal cell layer of CA regions, as well as in hilus and granular cell layer of dentate gyrus in hippocampus. Moreover, immunoblotting analysis showed that the expres- sion level of Muncl 8-1 in nucleus fraction of hippocampus significantly decreased in KA-treated animals. The relation- ship between the change of Muncl8-1 expression in neuronal nuclei and neuronal over-activation was also tested in pri- mary cultured neurons. After treatment with 50 ~tmol/L glutamate acid for 3 h, Muncl8-1 level was decreased in nucleus fraction and increased in cytoplasmic fraction of primary cultured neurons. Conclusion These results suggest that excit- atory stimulation can induce the distribution change of Munc 18-1 in neuron, which may subsequently modulate neuronal functions in brain.展开更多
Heat-stroke is a serious form of hyperthermia with high mortality, and can induce severe central nervous system disorders. The neurovascular unit(NVU), which consists of vascular cells, glial cells, and neurons, con...Heat-stroke is a serious form of hyperthermia with high mortality, and can induce severe central nervous system disorders. The neurovascular unit(NVU), which consists of vascular cells, glial cells, and neurons, controls blood-brain barrier(BBB) permeability and cerebral blood flow, and maintains the proper functioning of neuronal circuits. However, the detailed function of each BBB component in heat-stroke remains unknown. In order to interpret alterations caused by heat stress, we performed transcriptome comparison of neuron and astrocyte primary cultures after heat treatment. Differentially-expressed genes were then selected and underwent Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway analysis. Gene-act networks were also constructed, and the expression of pivotal genes was validated by quantitative PCR, as well as single-cell q PCR in heatstroke rats. Our work provides valuable information on the transcriptional changes in NVU cells after heat stress,reveals the diverse regulatory mechanisms of two of these cellular components, and shows that a cell-type-specificapproach may be a promising therapeutic strategy for heatstroke treatments.展开更多
文摘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, H202, testosterone, and testosterone (pre-added) plus H2O2 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 H202 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 H2O2 group were significantly greater than in the H2O2 group (P 〈 0.05), and nitric oxide synthase and malondialdehyde levels were significantly less than in the H2O2 group (P〈 0.05). CONCLUSION: Androgen partially reversed H2O2-induced neuronal damage and protected neurons.
文摘The effects of bis(7) tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate induced cell injury were investigated in primary cerebral cortical neurons of rats. Exposure of cultured neurons (12 days after plating) to 0.5 mmol/L glutamate for 30 min resulted in significant cell damage. Pretreatment with bis(7) tacrine (0.03 1.0 μmol/L) reduced the glutamate induced neurotoxicity in a concentration dependent manner and the maximal response was seen at 1 μmol/L with approximately 30% protection. A receptor binding assay showed that bis(7) tacrine can completely displace MK 801 binding to rat cortical membrane with an IC 50 of 0.57 μmol/L. These findings suggest that bis(7) tacrine can directly interact with N methyl D aspartate receptor channel complex, which may contribute to the inhibitor's protective effects against glutamate induced excitotoxicity. Thus, it is possible that anti glutamate/anti AChE synergism is responsible for potentially better Alzheimer's therapy of bis(7) tacrine relative to tacrine.
文摘Primary cultures of rat cerebral corticai neurons were prepared from conices or 2 days Sprague-Dawlley rats.We made an attempt to imitate the hypoxic neurons injury for testing neuroprotective erfect or Sibelium. It was found that Sibelium could significantly decrease the amounts of LDH leaking from intracellular fluid and increase the neuron-membrane fluidity of hypoxic neurons.Hypoxic group had significantly higher incorporation of 3H-TDR and 3H-UR than control group,while Sibelium could decrease the incorporation of 3H-TDR and 3H-UR. In addition,Sibelium could decrease the incorporation of 3H-TDR and 3H-UR in normal cultured neurons.The results showed that Sibelium could protect the neuronmembranes against alteration caused by hypoxia and regulate the synthesis of DNA and RNA.
基金supported by the National Natural Science Foundation(31170799 and 30872082)the National Basic Research Development Program(973 Program)of China(2011CB503702)
文摘The prevalence of domestic and industrial electrical appliances has raised concerns about the health risk of extremely low-frequency magnetic fields(ELF-MFs). At present, the effects of ELF-MFs on the central nervous system are still highly controversial, and few studies have investigated its effects on cultured neurons. Here, we evaluated the biological effects of different patterns of ELF-MF exposure on primary cultured hippocampal neurons in terms of viability, apoptosis, genomic instability,and oxidative stress. The results showed that repeated exposure to 50-Hz 2-mT ELF-MF for 8 h per day after different times in culture decreased the viability and increased the production of intracellular reactive oxidative species in hippocampal neurons. The mechanism was potentially related to the up-regulation of Nox2 expression.Moreover, none of the repeated exposure patterns had significant effects on DNA damage, apoptosis, or autophagy, which suggested that ELF-MF exposure has no severe biological consequences in cultured hippocampal neurons.
基金supported by grants from the National Natural Science Foundation of China (No. 81071017, 30470536, 90919004)
文摘Objective Muncl8-1 has an important role in neurotransmitter release, and controls every step in the exocy- totic pathway in the central nervous system. In the present study, whether epileptic seizure causes a change of Muncl8 localization in neuronal nuclei was analyzed. Methods Epilepsy models were established by injection of kainic acid (KA) solution into hippocampus of Sprague-Dawley (SD) rats or intraperitoneal injection of KA in Kunming mice. The hippocampal neurons were prepared from embryonic day 18 SD rats, and cultured in neurobasal medium, followed by treatment with glutamate for 3 h. Neuronal and glial nuclei of hippocampus were separated by sucrose density gradient centrifugation. The nucleus-enriched fractions were stained with 0.1% Cresyl Violet for morphological assay. Immuno- chemistry and immunoelectron microscopy with anti-Muncl 8-1 antibody were used to determine the nuclear locatization of Munc 18-1. Immunoblotting was used to detect the protein level of Munc 18-1. Results The localization of Munc 18-1 in nucleus of rat hippocampal neuron was confirmed by immunochemistry, immunoelectron microscopy, and immunob- lotting detection of neuronal nucleus fraction. In animals receiving intrahippocampal or intraperitoneal injection of KA, immunostaining revealed that the expression of Muncl 8-1 decreased in pyramidal cell layer of CA regions, as well as in hilus and granular cell layer of dentate gyrus in hippocampus. Moreover, immunoblotting analysis showed that the expres- sion level of Muncl 8-1 in nucleus fraction of hippocampus significantly decreased in KA-treated animals. The relation- ship between the change of Muncl8-1 expression in neuronal nuclei and neuronal over-activation was also tested in pri- mary cultured neurons. After treatment with 50 ~tmol/L glutamate acid for 3 h, Muncl8-1 level was decreased in nucleus fraction and increased in cytoplasmic fraction of primary cultured neurons. Conclusion These results suggest that excit- atory stimulation can induce the distribution change of Munc 18-1 in neuron, which may subsequently modulate neuronal functions in brain.
基金supported by the 12thFive-Year Plan of the PLA (BWS11J062)the China Postdoctoral Science Foundation (2015M572806)the Director’s Fund of the General Hospital of Jinan Military Region, China (2014ZX03)
文摘Heat-stroke is a serious form of hyperthermia with high mortality, and can induce severe central nervous system disorders. The neurovascular unit(NVU), which consists of vascular cells, glial cells, and neurons, controls blood-brain barrier(BBB) permeability and cerebral blood flow, and maintains the proper functioning of neuronal circuits. However, the detailed function of each BBB component in heat-stroke remains unknown. In order to interpret alterations caused by heat stress, we performed transcriptome comparison of neuron and astrocyte primary cultures after heat treatment. Differentially-expressed genes were then selected and underwent Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway analysis. Gene-act networks were also constructed, and the expression of pivotal genes was validated by quantitative PCR, as well as single-cell q PCR in heatstroke rats. Our work provides valuable information on the transcriptional changes in NVU cells after heat stress,reveals the diverse regulatory mechanisms of two of these cellular components, and shows that a cell-type-specificapproach may be a promising therapeutic strategy for heatstroke treatments.