摘要
目的研究AMP激活的蛋白激酶(AMPK)信号通路在蛛网膜下腔出血(SAH)大鼠脑组织中的表达,探讨AMPK参与SAH早期脑损伤中细胞凋亡的机制。方法血管内穿刺法建立SAH模型,免疫组化检测AMPK与磷酸化AMPK(p-AMPK)的组织细胞定位,RT-PCR及Western blot检测造模后6、24、48、72h额底皮质AMPK mRNA及p-AMPK、Bim、caspase-3蛋白的动态表达,并与正常对照组和假手术组比较。侧脑室给予AMPK激动剂AICAR及抑制剂Compound C,检测药物干预对大鼠神经行为及凋亡相关蛋白表达的影响,并与SAH组及vehicle组进行比较。结果 SAH后AMPKα的mRNA表达升高,p-AMPK、Bim、caspase-3蛋白表达均持续升高;AICAR可以增高AMPK磷酸化和Bim、caspase-3表达水平,加重神经功能缺损,Compound C可以减低AMPK磷酸化和Bim、caspase-3表达水平,改善神经功能评分。结论 AMPK信号通路参与了SAH后早期脑损伤中神经元细胞凋亡的病理生理过程,其机制可能与调控Bim的转录活性有关,抑制AMPK信号通路可以减轻SAH后皮层神经元凋亡。
Objective To investigate the activation of AMP-activated protein kinase （AMPK） and Bim in rat cortex after subarachnoid hemorrhage （SAH） and to explore the role of AMPK in cortical apoptosis in early brain injury after SAH. Methods SAH model was established by an endovascular perforation technique. The immunohistochemical method was used to detect the localization of AMPK and phosphorylated AMPK. RT-PCR was performed to detect the dynamic mRNA expression of AMPKa, and Western blot was used to detect the protein levels of phosphorylated AMPK and apoptosis-related proteins （Bim and caspase-3） in the cortex. After AICAR and compound C were administered via i. c. v, the change of p-AMPK level was examined, and the effects of AICAR and compound C on neurological behavior and apoptosis-related protein expressions were observed. Results Experimental SAH increased the expressions of AMPK~ mRNA and phosphorylated AMPK. Bim and caspase-3 were also significantly increased. With compound C treatment, the phosphorylation level of AMPK and the up-regulation of Bim were modified. However, AICAR administration exacerbated cortical apoptosis and neurological deficit through up-regulation of Bim. Conclusion AMPK is involved in the pathophysiological process of neuronal apoptosis in early brain injury after SAH. The mechanism may be related to the regulation of transcription of Bim. Inhibition of AMPK signaling pathway can provide neuroprotection by reducing cortical apoptosis after SAH.
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2013年第6期704-709,716,共7页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.30870844)
陕西省"13115"重大科技创新专项基金资助项目(No.2008ZDKG-66)
教育部高等学校博士学科点专项科研基金资助项目(No.20110201110060)~~
关键词
蛛网膜下腔出血
细胞凋亡
早期脑损伤
AMPK
BIM
大鼠
subarachnoid hemorrhage
apoptosis
early brain injury
AMP-activated protein kinase （AMPK）
Bim
rats
