吡格列酮对谷氨酸引起皮质神经元损伤的保护作用及其机制

Protection of pioglitazone on cultured cortical neurons from glutamate induced neurotoxicity via inhibiting p38 MAPK

目的观察吡格列酮对谷氨酸引起皮质神经元损伤的保护作用及机制。方法初生大鼠乳鼠的皮质神经元体外培养7 d后用于实验,建立谷氨酸损伤模型,用Hoechst33258核染色观察细胞凋亡的形态学改变,MTT法测定细胞活力,W estern印迹法检测磷酸化p38MAP激酶蛋白水平。结果谷氨酸组细胞存活率较对照组明显降低(P<0.01),吡格列酮、SB203580和一氧化氮合酶抑制剂(SMT)提高细胞存活百分率(P<0.01)。谷氨酸组细胞凋亡百分比较对照组增加(P<0.01),吡格列酮SB203580和SMT均能降低细胞凋亡百分比(P<0.01)。谷氨酸组磷酸化p38MAP激酶蛋白水平较对照组明显升高(P<0.01),吡格列酮明显降低磷酸化p38MAP激酶蛋白水平(P<0.01)。结论吡格列酮对谷氨酸引起体外培养皮质神经元损伤具有保护作用,此作用与降低磷酸化p38MAP激酶蛋白水平有关。

Objective To investigate whether pioglitazone protection effect on cultured cortical neurons injured by glutamate and the possible molecular mechanisms.Methods The cortical neurons were taken from new born rats and used for experiments 7 days after culture.The neurons were randomly divided into control;glutamate;glutamate ＋ pioglitazone;glutamate ＋ SB203580 and glutamate ＋ SMT groups.Western blotting was performed to investigate the protein level of phospho-p38MAPK.MTT was used to measure cell viability.The morphology change of neurons was observed under a fluorescence microscope with fluorescence dye Hoechst 33258.Results Pioglitazone markedly reduced the damage of cortical neurons induced by glutamate（P0.01） and significantly inhibited glutamate induced up-regulation of phospho-p38MAPK protein level（P0.01）.SB203580 and SMT antagonized the toxicity caused by glutamate to certain extent（P0.01）.Conclusions Pioglitazone can protect cultured cortical neurons from glutamate induced neurotoxicity. The protective effect of pioglitazone appears to be associated with inhibiting p38MAPK.