摘要
目的:观察蝎毒耐热蛋白(SVHRP)对红藻氨酸(KA)诱导的原代培养海马神经肽Y(NPY)能神经元损伤的影响及其可能的分子机制。方法:制备原代培养10d的大鼠海马神经元并用神经元特异性MAP-2抗体进行鉴定,将鉴定成熟的神经元用终浓度为20μg/ml的SVHRP和10μmol/L的KA处理,共孵育24h后,分别用硫堇染色、MTT实验检测不同给药组残存神经元的数目和活力,用免疫细胞化学和RT-PCR技术检测NPY-IR和NPYmRNA的表达。结果:MAP-2-IR结果显示85%以上为阳性成熟神经元;硫堇染色显示,同模型组比较,模型给药组未见神经元形态异常,并且神经元数目未见明显减少(P<0.05);MTT实验显示,模型给药组海马神经元存活率较模型组明显增高(P<0.05);NPY-IR检测表明,模型组NPY阳性细神经元数目明显减少,模型给药组NPY阳性神经元数目明显多于模型组(P<0.01);RT-PCR实验表明,单独给药组海马神经元内NPYmRNA表达较其他三组明显增多(P<0.05)。结论:SVHRP对KA诱导的原代海马神经元的兴奋毒性损伤具有明显的保护作用,可能与SVHRP促进NPY合成有关。
Aim：To investigate the effects of scorpion venom heat-resistant protein（SVHRP）on kainic acid induced-damage of cultured primitive rat hippocampal neuropeptide Y-nergic neurons.Methods：We observed morphological changes,celluar vigor,NPY-immunoreactivity and NPY mRNA expression by means of Thionine staining,MTT assay,immunocytochemistry and RT-PCR,respectively,on the primitively cultured Sprague-Dawley rat hippocampal neuron treated with KA and SVHRP for 24 h.Results：MTT assay and morphologic analysis showed that SVHRP markedly increased neuron survival-rate,and protected them from kA-induced damage.The expression of NPY-immunoreactivity and NPY mRNA in SVHRP group increased obviously compared with other groups.Conclusion：SVHRP protected the primitively cultured hippocampal neurons from KA-induced neuroexcitotoxicity and promoted the expression of NPY.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2007年第3期315-318,I0002,共5页
Chinese Journal of Applied Physiology
关键词
癫痫
蝎毒耐热蛋白
红藻氨酸
原代培养海马神经元
epilepsy
scorpion venom heat-resistant protein
kainic acid,primitively cultured hippocampal neuron
