低氧对SH-SY5Y细胞miR-124及BACE1蛋白表达的影响  

Effects of hypoxia on the expressions of miR-124 and BACE1 protein in SH-SY5Y cells

作　　者：张晓雯[1] 崔静[1] 李良[1] 

机构地区：[1]首都医科大学基础医学院病理系,北京100069

出　　处：《基础医学与临床》2015年第4期470-474,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(81070926;81271310)

摘　　要：目的研究低氧对培养细胞miR-124及β-淀粉样前体蛋白裂解酶1(BACE1)蛋白表达的影响。方法培养人神经母细胞瘤细胞(SH-SY5Y),分为对照组、氧-糖剥夺组、Aβ1-42处理组、过表达或抑制miR-124组,用实时定量PCR方法检测miR-124表达,Western blot法检测BACE1蛋白表达。结果氧-糖剥夺处理48 h,细胞内miR-124表达水平明显下降,仅为对照组的46.9%(P<0.05),BAE1蛋白表达水平与较对照组增高36%(P<0.01);转染miR-124过表达组组细胞内miR-124表达水平较对照组增高245倍(P<0.01),为miR-124过表达对照组的219倍(P<0.01),BACE1蛋白表达水平较对照组下降了29%(P<0.05),较miR-124过表达对照组下降25%(P<0.05)。转染miR-124抑制剂组miR-124表达水平下降至对照组的45.4%(P<0.05),至miR-124抑制对照组的48%(P<0.05),BACE1表达水平较对照组升高21%(P<0.05),较miR-124抑制对照组升高40.3%(P<0.01);细胞经Aβ1-42处理24 h,miR-124表达水平较对照组下降52%(P<0.05),BACE1蛋白表达水平较对照组增高51%(P<0.05)。结论低氧通过降低SH-SY5Y细胞miR-124表达进而升高BAEC1蛋白表达,且可能在阿尔茨海默病(Alzheimer's disease,AD)早期发病中发挥作用。Objective To investigate the role of hypoxia on the expression of miR-124 and BACE1. Methods Cultured SH-SY5Y cells were divided into blank control group, oxygen-glucose deprivation （OGD） group, Aβ_1-42 group and overexpression or inhibition of miR-124 group. RT-PCR was performed to analyze the expression of miR- 124 level. The level of BACE1 was examined by Western blot. Results After OGD for 48 h, the expression level of miR- 124 decreased significantly to 46.9% of the control group （ P 〈 0.05 ） , while the expression level of BACE1 was increased by 36% compared with the control group （P 〈 0.01 ）. After transfection of 48 h, miR- 124 level of miR-124 mimic group was increased by 245 folds as compared with control （P 〈 0.01 ） and by 219 folds as compared with miR-124 mimic control （P 〈0.01 ） ; BACE1 level was decreased by 29% compared with control （ P 〈 0.05 ） and by 25% compared with miR-124 mimic control （P 〈 0.05 ） ; miR-124 level of miR-124inhibitor group was decreased to 45.4% of control （ P 〈 0. 05 ）, 48% of miR- 124 inhibitor control （ P 〈 0. 05 ）, BACE1 level was increased by 21% compared with control （P 〈0.05） , 40. 3% compared with miR-124 inhibitor control （P 〈 0. 01 ）. miR-124 level was decreased by 52% compared with control （P 〈 0. 05） and the level of BACE1 level was increased by 51% after A[31.42 treatment for 24 h （P 〈 0. 05 ）. Conclusions Hypoxia may in- crease the expression of BACE1 through the clown-regulation of miR-124 which plays a role in the early pathogene- sis of Alzheimer＇s disease.

关 键 词：低氧 miR-124 β-淀粉样蛋白(AB) β-淀粉样前体蛋白裂解酶1(BACE1) 

分 类 号：R749.16[医药卫生—神经病学与精神病学]