蝎毒耐热蛋白对红藻氨酸诱导的海马NPY能神经元损伤的影响

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。

Effects of scorpion venom heat-resistant protein on kainic acid induced-damage of cultured primitive rat hippocampal neuropeptide Y-nergic neurons

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.