摘要
目的观察脑缺血再灌注中芳香族-L-氨基酸脱羧酶(AADC)对凋亡的作用和与PI3K/Akt信号通路的关系。方法建立tMCAO大鼠和OGD/RPC12细胞模型,检测大脑皮层神经元和PC12细胞凋亡情况与AADC、PI3K/Akt信号通路及下游凋亡相关蛋白的表达情况。结果体内实验结果显示,皮层神经元受损时,模型组AADC蛋白表达水平显著上升(P<0.01)。体外实验结果显示,PC12细胞存活率降低时,AADC蛋白表达水平显著上升(P<0.01),PI3K显著下降(P<0.01),Bcl-2等抗凋亡蛋白降低,Caspase-3等促凋亡蛋白升高。沉默AADC可促进PC12细胞存活并使PI3K、Akt等抗凋亡蛋白表达增加,Caspase-3等促凋亡蛋白表达降低。结论AADC在CIRI中可通过抑制PI3K/Akt信号通路,影响下游Bcl-2、Caspase-3等凋亡相关蛋白表达诱发神经元凋亡。
Objective To investigate the role of AADC in cerebral ischemia reperfusion injury on apoptosis and its relationship with the PI3K/Akt signaling pathway.Methods In this study,we established tMCAO rat model and OGD/R PC12 cell model,detecting the apoptosis of cortex neurons and PC12 cells and the expression of AADC,PI3K/Akt signaling pathway and downstream apoptosis related protein.At the same time,siRNA and LY294002 were used to silence or inhibit AADC and PI3K,and the expression of downstream apoptosis related proteins was observed under low expression conditions,to explore whether the relative expression changes were related to AADC.Results In vivo experimental results showed that when cortical neurons were damaged,the expression levels of AADC protein and mRNA in model group significantly increased.In vitro experimental results showed that when the survival rate of PC12 cells decreased,the expression level of AADC protein significantly increased(P<0.01),PI3K significantly decreased(P<0.01),anti apoptotic proteins such as Bcl-2 decreased,and pro apoptotic proteins such as Caspase-3 increased.Silencing AADC can promote the survival of PC12 cells and increase the expression of anti apoptotic proteins such as PI3K and Akt,while reducing the expression of pro apoptotic proteins such as Caspase-3.Conclusion This study indicates that AADC can induce neuronal apoptosis in CIRI by inhibiting the PI3K/Akt signaling pathway and affecting the expression of downstream apoptosis related proteins such as Bcl-2 and Caspase-3.
作者
路颖慧
尚津锋
黄贵金凤
文胤琏
张晓璐
刘欣
LU Ying-hui;SHANG Jin-feng;HUANG Gui-jin-feng;WEN Yin-lian;ZHANG Xiao-lu;LIU Xin(College of TCM,Beijing University of Chinese Medicine,Beijing 102488)
出处
《湖北中医药大学学报》
2023年第5期5-9,共5页
Journal of Hubei University of Chinese Medicine
基金
国家自然科学基金面上项目(项目编号:81573726)
北京中医药大学“解码中医”揭榜挂帅项目(项目编号:2022-JYB-JBZR-005)。
