调节神经元CREB磷酸化水平对APP和γ分泌酶的影响  

Effect of regulating the phosphorylation level of CREB on APP and secretaseγ

作　　者：邸畅 李国萍[2] 罗浩丹 毕珊 孙欣 陈绍春 DI Chang;LI Guo-Ping;LUO Hao-Dan(Department of Anatomy and Histoembryology,Kunming Medical University,Kunming 650500,Yunnan,China)

机构地区：[1]昆明医科大学基础医学院人体解剖与组织胚胎学系,云南昆明650500 [2]昆明医科大学第三附属医院头颈外科,云南昆明650500 [3]昆明医科大学康复学院,云南昆明650500

出　　处：《中国老年学杂志》2021年第8期1682-1686,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81302362);云南省科技厅-昆明医科大学联合专项基金(2018FE001);昆明医科大学科技创新团队项目(CXTD201905)。

摘　　要：目的在人神经元样细胞SY5Y中激活和抑制环磷酸腺苷(cAMP)反应元件结合蛋白(CREB)的磷酸化,检测γ分泌酶复合体蛋白及β淀粉样蛋白前体(APP)水平的变化,分析通过调控CREB磷酸化防治阿尔茨海默病(AD)的可行性。方法培养空载体SY5Y细胞(Vector-SY5Y)、过表达Gαq(Gαq-SY5Y)细胞、过表达APP(APP-SY5Y)细胞,分别用cAMP激动剂Forskolin、蛋白激酶(PK)A受体抑制剂H 892HCL处理上述3种细胞,Western印迹检测APP及γ分泌酶复合体的蛋白成分早老素增强子(PEN2)及早老素C端(PS1-CTF)的水平变化。结果①在Vector-SY5Y细胞中,激动剂组APP的表达无明显变化,PEN2、PS1-CTF表达明显上调;抑制剂组APP的表达明显上调,PEN2、PS1-CTF变化不明显。②在Gαq-SY5Y细胞中,激动剂组APP明显下降,PEN2表达量明显上调,PS1-CTF明显下降;抑制剂组APP明显下降,PEN2明显下调,PS1-CTF无明显变化。③在APP-SY5Y细胞中,激动剂组APP变化不明显,PEN2表达明显上调,PS1-CTF表达明显下降;抑制剂组APP明显上升,PEN2表达明显下降,PS1-CTF明显上调。结论激活CREB的磷酸化可上调γ分泌酶复合体中PEN2水平,协同过表达Gαq可明显降低APP水平,可设计基于CREB和Gαq的AD防治策略。Objective To research the affection of activation or inhibition of phosphorylation of cAMP-response element binding protein(CREB)on the levels of amyloidβprotein precursor(APP)andγ-secretase complex in neuron like cell lines SY5Y,then to analyze the feasibility of preventing and treating Alzheimer′s disease(AD)by regulating CREB.Methods Vector-SY5Y cells(transfected with only empty vector),Gαq-SY5Y cells(over-expression with Gαq)and APP-SY5Y cells(over-expression with APP)were treated with Forskolin(to activate CREB)and H 892HCL(to inhibit CREB)respectively.Western blot was applied to determine the level of APP andγ-secretase components progesterone enhancer 2(PEN2)and progesterone 1 c-terminal(PS1-CTF).Results①In CREB activated Vector-SY5Y cells,APP had no significant change,PEN2 and PS1-CTF were significantly upregulated;in CREB inhibited Vector-SY5Y cells,APP was significantly enhanced,levels of PEN2 and PS1-CTF had no significant change.②In CREB activated Gαq-SY5Y cells,APP was significantly decreased,PEN2 was significantly upregulated,PS1-CTF was significantly downregulated;in CREB inhibited Gαq-SY5Y cells,APP and PEN2 were significantly decreased,PS1-CTF had no significant change.③In CREB activated APP-SY5Y cells,APP had no significant change,PEN2 was significantly upregulated,level of PS1-CTF was significantly downregulated;in CREB inhibited APP-SY5Y cells,APP was significantly enhanced,PEN2 was significant decreased,PS1-CTF was significantly upregulated.Conclusions Activation of phosphorylation of CREB could upregulateγ-secretase components PEN2.CREB activation combined with Gaq overexpression could downregulate APP level.It is possible to design strategy of preventing and treating AD based on CREB and Gαq.

关 键 词：β淀粉样蛋白前体 Γ分泌酶 早老素1 早老素增强子2 环磷酸蛋白反应元件结合蛋白 

分 类 号：R34[医药卫生—基础医学]