阿托伐他汀对谷氨酸引起大鼠神经元损伤保护作用机制的研究

目的探讨阿托伐他汀对谷氨酸所致培养大鼠皮层神经元损伤保护作用的机制。方法 MTT法测定细胞存活率;Hoechst33258核染色观察细胞凋亡的形态学改变;Western blot检测活性的半胱氨酸天冬氨酸蛋白酶-3(caspase-3)和钙蛋白酶Ⅰ蛋白表达水平。结果谷氨酸(100μmol.L-1)可使神经元细胞存活率下降,细胞凋亡百分比明显增加,活性的caspase-3和钙蛋白酶Ⅰ蛋白表达增加。阿托伐他汀明显对抗谷氨酸诱导的神经元存活率下降及细胞凋亡百分比增加,同时明显抑制活性的caspase-3和钙蛋白酶Ⅰ蛋白表达增加。磷酯酰肌醇-3激酶(phos-phoinositide3-kinase,PI3K)/磷酸化蛋白激酶B(protein ki-nase B,Akt)通路特异性阻断剂LY294002(10μmol.L-1)能抑制阿托伐他汀对抗谷氨酸引起神经元细胞存活率下降,细胞凋亡百分比增加及活性caspase-3和钙蛋白酶Ⅰ蛋白表达增加的作用。结论阿托伐他汀能够明显对抗谷氨酸引起的皮层神经元损伤作用,这种作用可能与激活PI3K/Akt信号转导通路有关。

The mechanisms of neuroprotective effect of atorvastatin on glutamate-induced neurotoxicity in cultured cortical neurons of rats

Aim To investigate the possible mechanisms responsible for neuroprotective effect of atorvastatin against glutamate-induced neurotoxicity in cultured cortical neurons of rats.Methods The cell viability was assessed by MTT assay.The morphology change of neurons was observed under a fluorescence microscope with fluorescence dye Hoechst33258.The protein expression of active caspase-3 and μ-calpain was measured by Western blot.Results The decrease of cell viability and the increase of apoptotic cells in cultured cortical neurons were observed when incubated with glutamate for 24 h compared with the normal controls.Glutamate induced the increases of active caspase-3 and μ-calpain protein expression.Atorvastatin inhibited glutamate-induced decrease in cell viability and increase of apoptotic cells,as well as increases of active caspase-3 and μ-calpain protein expression.PI3K/Akt inhibitor LY294002(10 μmol·L-1)inhibited the afore-mentianed effects of atorvastatin.Conclusion We demonstrate that atorvastatin can protect cultured cortical neutons of rats from glutamate-induced damage,and atorvastatin protects cortical neurons against glutamate insult through activating PI3K/Akt pathway.