ERK1/2和PI3-K通路抑制剂对蛛网膜下腔出血大鼠海马区神经细胞自噬的协同作用  

Synergistic Effects of ERK1/2 and PI3-K Pathway Inhibitors on Autophagy in the Hippocampus of Rats with Subarachnoid Hemorrhage

作　　者：安朝旺 刘瑶 赵晓云 韩颖 赵雅宁 赵旭 李建民 薛承景 

机构地区：[1]华北理工大学附属医院神经外科,河北唐山063000 [2]华北理工大学护理与康复学院临床教研室,河北唐山063000

出　　处：《中国医科大学学报》2017年第11期995-1000,共6页Journal of China Medical University

基　　金：河北省省级重大医学科研课题(zd2013087);唐山市科技计划(14130220B)

摘　　要：目的通过观察联合抑制细胞外信号调节激酶(ERK1/2)和磷脂酰肌醇3激酶(PI3-K)蛋白对神经细胞自噬的影响,探讨蛛网膜下腔出血(SAH)救治靶点。方法将200只雄性SD大鼠按照数字表法随机分为假手术(Sham)组、模型(SAH)组、ERK1/2抑制剂U0126组、PI3-K抑制剂LY294002组、U0126+LY294002组。二次注血法制作SAH大鼠模型;HE染色观察海马区神经细胞的形态变化;实时荧光定量PCR法检测ERK1/2、PI3-K、自噬相关蛋白beclin-1和微管相关蛋白1轻链(LC3)m RNA的表达,免疫组化法检测海马区磷酸化ERK1/2、PI3-K、beclin-1和LC3-Ⅱ蛋白的表达水平。结果 SAH组海马区神经细胞死亡率、ERK1/2、PI3-K、beclin-1和LC3 m RNA以及磷酸化ERK1/2、PI3-K、beclin-1和LC3-Ⅱ蛋白水平均高于Sham组(均P<0.05);U0126组和LY294002组海马区神经细胞死亡率高于SAH组,而ERK1/2、PI3-K、beclin-1和LC3 m RNA以及磷酸化ERK1/2、PI3-K、beclin-1和LC3-Ⅱ蛋白水平均低于SAH组(均P<0.05);U0126+LY294002组海马区神经细胞死亡率高于U0126或LY294002组,而ERK1/2、PI3-K、beclin-1和LC3 m RNA以及磷酸化ERK1/2、PI3-K、beclin-1和LC3-Ⅱ蛋白水平分别低于U0126和LY294002组(均P<0.05)。结论联合抑制ERK1/2和PI3-K通路激活可显著降低SAH后神经细胞自噬,加重神经细胞丢失。Objective To investigate synergistic effects of extracellular signal regulated kinase （ERK1/2） and phosphatidylinositol 3 kinase （PI3-K） pathway inhibitors on autophagy in the hippocampus of rats with subarachnoid hemorrhage （SAH）,for identification of therapeutic targets in SAH. Methods Totally, 200 male SD rats were randomly divided into a sham operated group, SAH group,inhibitor U0126 group, inhibitor LY294002 group, and a U0126＋ LY294002 group. Animal models were established by injecting autologous blood twice into the eisterna magna. Morphological changes in the hippocampus nerve cells were detected by HE staining;ERK1/2 ,PI3-K,bee- 1in-1, and LC3 mRNA expression in the hippocampus were detected by real-time PCR, and phosphorylated ERK1/2, PI3-K, beclin-1, and LC3-Ⅱ protein expression were detected by immunohistochemistry. Results Neuronal death rate and phosphorylated ERK1/2,PI3-K, beclin-1, and LC3- Ⅱ levels in the hippocampus in the SAH group were higher than in the sham group （all P 〈 0.05）. Neuronal death rate in U0126 or LY294002 group was higher than in SAH group, while ERK1/2 ,P13-K,becliu-1, and LC3 mRNA and phosphorylated ERK1/2, PI3-K, beclin-1, and LC3- Ⅱ protein levels were lower than in SAH group （all P 〈 0.05）. Neuronal death rate in U0126 ＋LY294002 group was higher than in U0126 or LY294002 group,while ERK1/2 ,PI3-K,beclin-1 ,LC3 mRNA and phosphorylated ERK1/2,PI3-K,beclin-1, and LC3- Ⅱ protein levels in the hippocampus were lower than in U0126 or LY294002 group （all P 〈 0.05）. Conclusion Co-targeting the inhibition of ERK1/2 and PI3-K pathways can significantly reduce neuronal cell autophagy and aggravate cells loss after SAH.

关 键 词：蛛网膜下腔出血 细胞外信号调节激酶 磷脂酰肌醇3激酶 自噬相关蛋白 微管相关蛋白1轻链 

分 类 号：R651.1[医药卫生—外科学]