抑制P13刚Akt／mTOR信号通路对血管性痴呆大鼠海马CA1区自噬相关蛋白表达的影响  

Effect of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway inhibition on autophagy related protein expressions in CA1 area of hippocampus of vascular dementia rats

作　　者：王一超 刘斌 毛文静 马原源 任伯 邓春颖 贺永贵 吕超男 

机构地区：[1]华北理工大学临床医学院实验中心,唐山063000 [2]华北理工大学附属医院神经内一科,唐山063000

出　　处：《中华神经医学杂志》2017年第2期127-132,共6页Chinese Journal of Neuromedicine

基　　金：河北省重大医学科研课题（Zd2013087）;河北省高等学校科学技术研究重点项目（ZH2012046）

摘　　要：目的探讨抑制磷脂酰肌醇3激酶（P13K）／蛋白激酶B（Akt）／雷帕霉素靶蛋白（roTOR）信号通路对血管性痴呆rvD）大鼠海马CA1区自噬相关蛋白表达的影响。方法108只SD大鼠按随机数字表法分为假手术组、VD组、P13K抑制剂LY294002组（LY294002组1和mTOR抑制剂雷帕霉素组（雷帕霉素组），每组再依据取材时间不同分为1、2、4、8周4个时间点。后3组采用改良Pulsinelli四血管阻断法制备大鼠VD模型，后2组在制备VD模型前侧脑室注射10mmol／L LY294002溶液10μL或8ng／μL雷帕霉素4μL。分别在各时间点采用Westem blotting检测大鼠海马CA1区Beclinl蛋白表达及微管相关蛋白轻链3（LC3）Ⅱ／LC3Ⅰ比值，在4周时间点采用免疫荧光染色检测大鼠海马CA1区LC3表达。结果（11与假手术组比较，VD组各时间点Beclinl蛋白表达及LC3II／LC3I比值均增高，差异均有统计学意义（P〈0．05）；与VD组比较，LY294002组和雷帕霉素组各时间点Beclinl蛋白表达及LC3IIFLC3I比值均明显增高，差异均有统计学意义（氏0．05）。（2）免疫荧光染色结果显示：4周时，假手术组神经元胞质内LC3呈散在低表达，VD组LC3表达增多．LY294002组和雷帕霉素组LC3表达明显增多。结论抑制P13K／Akt／mTOR信号通路能促进VD大鼠海马CA1区自噬相关蛋白Beclinl及LC3表达，神经元自噬得到明显增强。Objective To investigate the effect ofphosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway on expressions of autophagy related proteins in CA1 area of hippocampus in vascular dementia （VD） rat models. Methods One hundred and eight healthy male SD rats were randomly divided into sham-operated group, VD group, PI3K inhibitor LY294002 group （LY294002 group） and roTOR inhibitor rapamycin group （rapamycin group, n=27）; the VD models in the later three groups were prepared by modified Pulsineli＇ s four-vessel occlusion; lateral ventricles in the later two groups were injected 10 mmol/L LY294002 solution 10 μL or 8 ng/μL rapamycin 4 μL before preparing of VD models. One, 2, 4 and 8 weeks after the success of model preparation, the autophagy associated protein Beclinl expression and microtubule-associated protein light chain （LC）3Ⅱ/LC3Ⅰ ratio in CA1 area of the hippocampus were detected by Western blotting. The expression of LC3 was detected in CA1 area of the hippocampus by fluorescence microscope 4 weeks after the success of model preparation. Results （1） As compared with those in the sham-operated group, the Beclinl expression and LC3Ⅱ/LC3Ⅰ ratio in VD group were significantly increased at all observation time points （P〈0.05）; As compared with those in the VD group, the Beclinl expression and LC3Ⅱ/LC3Ⅰ ratio in the LY294002 group and rapamycin group were significantly increased at all observation time points （P〈0.05）. （2） At 4 weeks after the success of model preparation, immunofluorescence results showed that LC3 in the cytoplasm of neurons was scattered with low expression in the sham-operated group, the LC3 expression was increased in the VD group, and that in the LY294002 group and rapamycin group was obviously increased. Conclusion Inhibition of PI3K/Akt/mTOR signaling pathway can promote the expressions of autophagy related protein Beclinl and LC3 in CA1 area of the hippocampus of vascul

关 键 词：血管性痴呆 磷脂酰肌醇3激酶 蛋白激酶B 雷帕霉素靶蛋白 细胞自噬 BECLIN1 微管相关蛋白轻链3 

分 类 号：R749.13[医药卫生—神经病学与精神病学]