FKBP51·PHLPP·Akt信号模块对大鼠脑缺血／再灌注后Akt磷酸化及海马神经元损伤的影响  

Effect of the FKBP51 · PHLPP·Akt signal module on the phosphorylation of Akt and hippocampal neu- ronal injury after the cerebral ischemia/reperfusion

作　　者：魏秀娥[1] 张风玉[2] 王凯[1] 张清秀[1] 荣良群[1] 

机构地区：[1]徐州医学院第二附属医院神经内科,徐州市221006 [2]聊城市人民医院神经内科

出　　处：《中华行为医学与脑科学杂志》2014年第1期15-18,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：徐州市科技局立项课题（XF10C030,2010）

摘　　要：目的探讨FKBP51·PHLPP·Akt信号模块对大鼠脑缺血／再灌注（I／R）后Akt磷酸化及海马CAl区迟发性神经元损伤的影响。方法采用Pulsinelli-Brierley法制作全脑缺血损伤模型，每亚组6只动物，缺血前连续3d侧脑室注射PH结构域且富含亮氨酸重复基序的丝／苏氨酸蛋白磷酸酶2（PHdo．mainand Leucinerichrepeat Protein Phosphatases，PHLPP2）反义寡核苷酸（antisense oligodeoxynucleotides，ASODN）后缺血15rain，再灌注6h，应用免疫共沉淀及免疫印记分别检测PHLPP2、FK506连接蛋白5（FK506bindingprotein5，FKBP51）和蛋白激酶B（ProteinkinaseB，Akt）三者两两结合、检测Akt及其磷酸化的表达情况。再灌注5d，断头取脑，石蜡切片，苏木精一伊红染色，图像分析测定单位面积内染色细胞面积总和。I／R＋ASODN组分别与I／R组及I／R＋错义寡核苷酸（missense0ligodeoxynucleotides，MSODN）组比较，进行统计学分析。结果单因素方差分析结果显示：与I／R＋PHLPP2MSODN组（1．24＋0．24）比较，I／R＋PHLPP2ASODN组（1．06±0．01）PHLPP2与Akt间的两两结合水平明显受到抑制（P〈0．05）；与I／R＋PHLPP2MSODN组（1．68±0．11）比较，I／R＋PHLPP2ASODN组（1．04±0．13）FKBP51与Akt间的两两结合水平明显受到抑制（P〈0．05）；与I／R＋PHLPP2MSODN组（0．58±0．01）比较，I／R＋PHLPP2ASODN组（0．76±0．02），P-Akt表达水平明显增加（P〈0．05），而Akt总蛋白表达无明显变化（P〉0．05）；I／R5d＋PHLPP2ASODN组[（88-3±2．7）个]与I／R＋PHLPP2MSODN组[（20．1±2．5）个]比较，差异有统计学意义（P〈0．05）。结论FKBP51·PHLPP·Akt模块可能通过Akt信号通路参与脑缺血再灌注损伤，促进海马CAl区神经元的迟发性损伤。Objective To investigate the effects of the FKBP51 · PHLPP · AKT signal module on the phosphorylation of Akt and hippocampal neuronal injury after the cerebral ischemia / repeffusion induced neuronal death in rat hippocampus. Methods Transient（ 15 min） brain ischemia was induced by the four-vessel occlusion in Sprague-Dawley rats. 6 rats were used in each group. The antisense oligodeoxynucletides（ AS ODN）of PHLPP2 （PH domain and leucine rich repeat protein phosphatases） was used to suppress the assembly of FKBP51 · PHLPP · Akt signal module by intracerebroventricular infusion once per day for 3 days before ischemia. After 6 hours reperfusion, interactions of PHLPP2 and FKBP51 （FK506 binding protein 5） wiih Akt were detected by im- munoprecipitation （IP） and the phosphorylation of Akt was detected by western blot （IB）. After 5 days reperfu- sion, rats were perfusion-fixed with paraformaldehyde and Hematoxylin-Eosin staining was used to examine the sur- vival number of CA1 pyramidal cells of hippocampus. Results Compared to PHLPP2 MS ODN group（ 1.24± 0.24,1.68±0.11,0.58±0.01）, PHLPP2 AS ODN suppressed the assembly of the FKBP51 · PHLPP·Akt signa- ling module（ 1.06±0.01, 1.04±0.13）, and increased the phosphorylation of Akt（0.76±0.02）（P〈0.05）. Further- more,compared to PHLPP2 MS ODN group （20.1±2.5） ,the number of surviving neurons significantly increased in PHLPP2 AS ODN group（88.3±2.7） （P〈0.05）. Conclusion The increasing assembly of FKBP51· PHLPP · Akt signal nxxlule can damage CA1 pyramidal cells of hippocampus by inhibiting the phosphorylation level of Akt.

关 键 词：全脑缺血 PH结构域且富含亮氨酸重复基序的丝 苏氨酸蛋白磷酸酶2 FK506连接 蛋白5 蛋白激酶B 

分 类 号：R743.31[医药卫生—神经病学与精神病学]