左归丸加减方对谷氨酸诱导的体外培养大鼠小脑颗粒神经元凋亡影响

Zuogui Pill Protects Rat Cerebellar Granule Neurons From Apoptosis Induced by Glutamate

目的 :研究左归丸加减方对谷氨酸引起的体外培养的小脑颗粒神经元 (CGNs)兴奋毒性死亡是否有抑制作用及其可能的分子机制。方法 :采用二乙酸荧光素 (PDA)和Hoechst332 5 8DNA染色法 ,观察神经元存活率及形态学特征 ,用琼脂糖凝脉电泳分析神经元死亡的生化特征。结果 :30 0 μmol·L- 1 谷氨酸可引起体外培养的CGNs调亡 ,左归丸加减方水提物 (ZGP)可以剂量依赖地阻断谷氨酸所致的CGNs调亡。PD980 5 9可以阻断谷氨酸的诱导CGNs调亡作用 ,协同ZGP的抗谷氨酸诱导CGNs调亡作用。LY2 94 0 0 2可以阻断ZGP的抗谷氨酸诱导CGNs调亡作用。结论 :ZGP对谷氨酸诱导的体外培养的大鼠CGNs调亡有保护作用 ,PI3 K Akt信号传导通路可能参与其作用。

OBJECTIVE To study whether ZGP can prevent cerebellar granule neurons from apoptosis induced by glutamate and its possible molecular mechanisms. METHODS The neurons were stained with fluorescein diacetate to observe their survival rate or stained with Hoechst 33258 to analyze their nuclear morphology. Internucleosomal DNA fragmentation of neurons was analyzed with agarose gel electrophoresis. RESULTS ZGP could increase the survival rate of cultured CGNSs in the medium containing 300μmol·L -1 glutamate concentration-dependently, and could prevent the neuronal nuclei from shrinkage, condensation and cleavage and nuclear DNA fragmentation induced by glutamate. LY294002 suppressed the protective effect of ZGP, PD98059 was able to block the excitotoxin of glutamate. CONCLUTION ZGP protected rat CGNs from appoptosis induced by glutamate and this effect was postulated to dependent PI3-K/Akt signaling pathway.