依达拉奉对海人酸诱导癫痫大鼠海马神经元损伤的保护作用

Protective effect of edaravone against neuron injury in KA-induced seizure of rats

目的:研究依达拉奉对海人酸诱导大鼠癫痫引起神经元损伤的保护作用及其分子机制。方法:SD大鼠随机分为癫痫对照组(saline组)、癫痫组(KA组)和药物对照组(KA+saline组)及依达拉奉组(KA+Ed组)。采用脑室注射海人酸制作大鼠癫痫模型。取海马组织应用Western-blot法检测cleaved caspase-3、FasL和Fas的表达,以免疫共沉淀法检测FasL与Fas的结合;焦油紫染色来观察海马CA1区CA3/DG区锥体细胞形态并计数成活的细胞。结果:癫痫组和药物对照组FasL的表达及其与Fas的结合以及cleaved caspase-3的表达较癫痫对照组显著增加(P<0.05),而依达拉奉组较药物对照组显著降低(P<0.05),Fas的蛋白表达各组间差异未见显著性(P>0.05);依达拉奉明显改善海人酸诱导损伤的神经细胞形态,减少CA1区CA3区神经元的缺失(P<0.05)。结论:依达拉奉可以通过抑制KA诱导FasL的表达及其与Fas的结合,抑制Fas/FasL通路,进而抑制caspase-3的活化对神经细胞发挥保护作用。

OBJECTIVE To investigate the neuroprotection of edaravone against neuronal death induced by kainic acid and to elucidate the molecular mechanisms. METHODS Adult male Sprague-Daley rats weighing 200-300 g were allotted into 4 groups as follows： control group （saline）, seizure group （KA）, drug control group （KA ＋ saline） , and edaravone group （Ed ＋ KA）. Seizures were induced by intracerebroventricular injection of KA dissolved in sterile saline. Edaravone was intraperitoneally administrated to the rats 40 min before KA injection. Subsequently a series of methods including immunoprecipitation,immunoblotting,histologic analysis were used to analyze the interaction,expression of relevant proteins as well as the survival of the CA1/CA3 pyramidal neurons. RESULTS Neuroprotective effect of edaravone was observed on neuronal injury induced by KA to hippocampal CA1 and CA3 pyramidal neurons. Edaravone could inhibit the expression of FasL and the combination of FasL with Fas and inhibit the cleavage of the caspase-3 zymogen to its active subunits induced by kainic acid in hippocampus. CONCLUSION Neuroprotective effect of edaravone may involve in inhibiting the expression of FasL and the combination of FasL with Fas and subsequently the activation of caspase-3 by decreasing the cleavage of the caspase zymogen to its active subunits.