ApoE通过影响mir-30a-5p调控BDNF表达  

Apo E regulates BDNF expression by influencing mir-30a-5p

作　　者：臧贵勇[1] 潘开昌[1] 龙友国[2] 余跃生[3] 李成朋 官志忠[4,5] 禹文峰[4,5] ZANG Guiyong;PAN Kaichang;LONG Youguo(Anatomy and Tissue Embryology Teaching and Research Section(Qiannan Medical School for Nationalities),Duyun 558000,Chian)

机构地区：[1]黔南民族医学高等专科学校解剖学与组织胚胎学教研室,贵州都匀558000 [2]黔南民族医学高等专科学校生物化学教研室,贵州都匀558000 [3]黔南民族医学高等专科学校预防医学教研室,贵州都匀558000 [4]地方病与少数民族疾病教育部重点实验室(贵州医科大学),贵州贵阳550004 [5]贵州医科大学分子生物学重点实验室,贵州贵阳550004

出　　处：《中风与神经疾病杂志》2018年第10期893-896,共4页Journal of Apoplexy and Nervous Diseases

基　　金：贵州省地方病和少数民族疾病创新人才团队(贵州省教育厅);黔南民族医学高等专科学校校基金;教育部"长江学者和创新团队发展计划资助"(IRT13058);贵州省科技计划[黔科合重大专项字(2014)6008号];贵州省创新计划项目[黔教合协同创新中心(2014)06];贵州省科技创新人才团队[黔科通(2016)161号]共同资助

摘　　要：目的研究ApoE基因型是如何影响脑源性神经营养因子(brain derived neurotrophic factor,BDNF)的表达,以更好的理解AD的发病机制。方法在SH-SY5Y细胞系和大鼠原代神经元中加入Apo E3和Apo E4重组蛋白,用real-time PCR、Western blot和ELISA检测BDNF的表达水平;用luciferase assay、real-time PCR和Western blot证实BDNF是mir-30a-5p的靶基因;用Taq Man技术检测mir-30a-5p的水平。结果与对照组相比,Apo E3增加了BDNF的mRNA、蛋白水平和培养基中的BDNF浓度,而ApoE4减少了BDNF的mRNA、蛋白水平和培养基中的BDNF浓度; hsa-mir-30a-5p抑制BDNF的3’UTR的luciferase活性,mRNA和蛋白水平。ApoE3降低mir-30a-5p的表达,Apo E4增加mir-30a-5p的表达,而过表达mir-30a-5p inhibitor可以逆转ApoE4对BDNF的抑制效果。结论BDNF是mir-30a-5p的靶基因,ApoE是通过影响mir-30a-5p的水平来调控BDNF的表达。Objective To investigate how ApoE genotype affects brain derived neurotrophic factor(BDNF)expression in the pathogenesis of AD.Methods ApoE3 and ApoE4 recombinant proteins were added to SH-SY5Y and rat primary neurons,the expression of BDNF was detected by real-time PCR,Western blot and ELISA.The luciferase assay,real-time PCR and Western blot confirm that BDNF is the target gene of mir-30a-5p.Results Compared to the control group,ApoE3 increased both BDNF mRNA and its protein levels as well as BDNF concentration in the culture medium while ApoE4 reduced BDNF at mRNA and protein levels as well as BDNF concentrations in the culture medium.At the same time,hsa-mir-30a-5p inhibits luciferase activity,mRNA and protein levels of the 3’UTR of BDNF.ApoE3 increased BDNF expression by decreasing mir-30a-5p,while overexpression of mir-30a-5p mimic reversed the effect of ApoE3 on BDNF.ApoE4 decreased BDNF expression by increasing mir-30a-5p,while overexpression of mir-30a-5p inhibitor reversed the inhibitory effect of ApoE4 on BDNF.Conclusion BDNF is a target gene for mir-30a-5p,and ApoE regulates the expression of BDNF by affecting the level of mir-30a-5p.

关 键 词：阿尔茨海默病 APOE 神经营养因子 mir-30a-5p 

分 类 号：R749.1[医药卫生—神经病学与精神病学]