Department of Neurology, Beijing Luhe Hospital Affiliated to Capital Medical University, Beijing, 101100, China.
Curr Neurovasc Res. 2019;16(5):473-480. doi: 10.2174/1567202616666191111124516.
Propose: To investigate whether miR-22-3p is able to regulate AD development and its molecular mechanism.
Morris water maze test was performed to test the spatial memory. Quantitative polymerase chain reaction (qPCR) was used to assess the expression level of miR-22-3p. The enzymelinked immunosorbent assay (ELISA) was used to assess the levels of Aβ40 and Aβ42. Immunoblotting analysis was performed to detect the protein expression levels of amyloid precursor protein (APP), mitogen-activated protein kinase 14 (MAPK14) and beta-site Amyloid precursor protein Cleaving Enzyme 1 (BACE1). Luciferase assay was used to identify the interaction between miR- 22-3p and MAPK14. The tetrazolium dye (MTT) colorimetric assay was used to test the influence of miR-22-3p overexpression on cell viability. Flow cytometry analysis was performed to evaluate the effect of miR-22-3p overexpression on cell apoptosis.
Morris water maze test showed that mice model of AD had impaired spatial memory, which was able to be ameliorated by miR-22-3p overexpression. Immunoblotting analysis revealed that the protein expression levels of APP, MAPK14 and BACE1 were enhanced in AD model, which could be prevented by miR-22-3p overexpression. ELISA showed that Aβ40 and Aβ42 levels were dramatically increased in AD model, which were inhibited by miR-22-3p overexpression. Luciferase assay and immunoblotting analysis indicated that miR-22-3p targeted and regulated MAPK14 expression.
MiR-22-3p overexpression reduced Aβ deposit and alleviated AD symptoms by targeting and regulating MAPK14 expression, which ameliorated AD symptoms.
探讨 miR-22-3p 是否能够调节 AD 的发生发展及其作用机制。
采用 Morris 水迷宫实验检测空间记忆能力，实时定量聚合酶链式反应（qPCR）检测 miR-22-3p 的表达水平，酶联免疫吸附测定（ELISA）检测 Aβ40 和 Aβ42 水平，免疫印迹分析检测淀粉样前体蛋白（APP）、丝裂原活化蛋白激酶 14（MAPK14）和β 位淀粉样前体蛋白裂解酶 1（BACE1）的蛋白表达水平，荧光素酶报告基因实验鉴定 miR-22-3p 与 MAPK14 的靶向关系，四唑盐（MTT）比色法检测 miR-22-3p 过表达对细胞活力的影响，流式细胞术检测 miR-22-3p 过表达对细胞凋亡的影响。
Morris 水迷宫实验结果显示 AD 模型小鼠的空间记忆受损，过表达 miR-22-3p 可改善 AD 模型小鼠的空间记忆能力。免疫印迹分析结果显示，AD 模型中 APP、MAPK14 和 BACE1 的蛋白表达水平升高，过表达 miR-22-3p 可抑制 AD 模型中 APP、MAPK14 和 BACE1 的蛋白表达水平。ELISA 结果显示，AD 模型中 Aβ40 和 Aβ42 水平显著升高，过表达 miR-22-3p 可降低 AD 模型中 Aβ40 和 Aβ42 水平。荧光素酶报告基因实验和免疫印迹分析结果表明，miR-22-3p 可靶向调控 MAPK14 的表达。
过表达 miR-22-3p 通过靶向调控 MAPK14 表达减轻 AD 症状，减少 Aβ 沉积，改善 AD 症状。
