甘草酸通过抑制自噬对脑缺血再灌注后神经元损伤的保护作用  

Neuroprotective Effect of Glycyrrhizic Acid in Rats with Cerebral Ischemia-reperfusion through Inhibition of Autophagy

作　　者：张印松 李光杰 相毅 ZHANG Yinsong;LI Guangjie;XIANG Yi(Department of Neurosurgery,Hengshui People's Hospital,Hengshui,Hebei,053000,China)

机构地区：[1]衡水市人民医院神经外科,河北省衡水市053000

出　　处：《医学分子生物学杂志》2023年第3期227-231,共5页Journal of Medical Molecular Biology

基　　金：衡水市科技计划项目(No.20220140582)。

摘　　要：目的探究甘草酸对脑缺血再灌注神经元损伤的保护作用.方法通过在大鼠原代神经元中进行氧糖剥夺再灌注(oxygen glucose deprivation reperfusion,OGD/R)构建体外脑缺血再灌注模型.显微镜观察细胞形态学变化,免疫荧光染色观察神经元特异性烯醇化酶(neuron-specific enolase,NSE)的表达.使用甘草酸对损伤神经元进行治疗,CCK-8法检测神经元细胞的活力变化,乳酸脱氢酶(LDH)-细胞毒性检测试剂盒检测神经元释放的LDH水平.使用AMPK激活剂AICAR对神经元AMPK/mTOR通路进行激活,Western印迹检测AMPK/mTOR通路相关蛋白及自噬相关蛋白的表达.结果甘草酸抑制OGD/R诱导的神经元LDH水平升高、细胞活力下降及自噬相关蛋白Beclin 1、LC3-Ⅱ的增加,并抑制AMPK/mTOR通路的激活.结论甘草酸介导AMPK/mTOR通路抑制神经元自噬从而对脑缺血再灌注后神经元发挥保护作用.Objective To investigate the protective effect of glycyrrhizic acid on neurons in rats with cerebral ischemia/reperfusion.Methods An cerebral ischemia-reperfusion model was constructed in rat primary neurons in vitro by oxygen glucose deprivation reperfusion(OGD/R)method.The changes of cell morphology were observed by microscope,and the expression of neuron-specific enolase(NSE)was observed by immunofluorescence staining.The damaged neurons were treated with glycyrrhizic acid,and the CCK-8 method was performed to detect the viability of neuronal cells,and the level of LDH released from neurons was detected by using the lactate dehydrogenase(LDH)-cytotoxicity assay kit.The AMPK activator AICAR was used to activate the AMPK/mTOR pathway in neurons,and the Western blotting assay was used to detect the expression levels of AMPK/mTOR pathway-related proteins and autophagy-related proteins.Results Glycyrrhizic acid inhibited the OGD/R-induced enhancement of neuronal LDH level and the expression levels of autophagy-related proteins Beclin 1 and LC3-II,and the suppression of cell viability.The OGD/Rinduced activation of AMPK/mTOR pathway was also suppressed after glycyrrhizic acid treatment.Conclusion Glycyrrhizic acid exerts a protective effect on neurons in rats with cerebral ischemia-reperfusion by inhibiting neuronal autophagy through AMPK/mTOR pathway.

关 键 词：脑缺血再灌注 氧糖剥夺再灌注 甘草酸 自噬 AMPK/mTOR 神经元保护 

分 类 号：R743.3[医药卫生—神经病学与精神病学]