MiR-615-5p缓解Aβ25-35诱导的APOE4＋/-型海马细胞的凋亡和变性

MiR-615-5p alleviates the apoptosis and degeneration of Aβ25-35-induced APOE4 + /-hippocampus cells

目的探讨miR-615-5p对人海马细胞的凋亡和变性的影响及潜在机制。方法运用实时定量PCR检测正常、家族性AD和散发性AD老年人血液中miR-615-5p的表达水平,以及APOE4+/-型人胚海马细胞和Aβ诱导的APOE4+/-海马细胞中miR-615-5p的表达水平;在Aβ预处理的APOE4+/-型海马细胞中分别转染miR-615-5p mimic和inhibitor后检测细胞凋亡、氧化应激标志物含量和突触生长标志蛋白的表达水平;运用生物学信息分析和荧光素酶报告基因技术预测并验证mir-615-5p对APOE基因的靶定调控关系。结果散发性AD患者和细胞模型中miR-615-5p的表达水平显著下调;过表达miR-615-5p显著抑制海马细胞的凋亡和细胞内氧化应激,并促进细胞内突触生长蛋白的表达,而沉默miR-615-5p则作用相反;mir-615-5p可靶定结合APOE mRNA,并下调APOE4+/-海马细胞中APOE4蛋白的表达。结论 mir-615-5p通过靶向调节APOE,缓解Aβ25-35诱导的APOE4+/-型海马细胞的凋亡和变性。

Objective Apolipoprotein E4（ apo E4） to explore the regulatory role and mechanism of miR-615-5p in the apoptosis and function of human hippocampus cells. Methods Expression of blood miR-615-5p were detected with real-time q PCR in familial Alzheimer＇s disease（ AD） patients,sporadic AD patients and non-AD aged volunteers,as well as APOE4 ＋ /- human embryonic hippocampus cells and Aβ-pretreated APOE4 ＋ /-hippocampus cells; the mir-615-5p mimic or inhibitor was transfected into the Aβ-pretreated APOE4 ＋ /-hippocampus cells,and then cell apoptosis,oxidative stress level and synaptic growth protein expression were detected; bioinformatics and luciferase reporter gene analyses were applied to evaluate and validate the targeting relationship between miR-615-5p and APOE4. Results MiR-615-5p was significantly decreased in sporadic AD patients and Aβ-pretreated APOE4 ＋ /-hippocampus cells; mir-615-5p mimic transfection suppressed cell apoptosis and oxidative stress,and promoted the expression of synaptic growth proteins; in contrast,mir-615-5p inhibitor transfection had an opposite effect; APOE mRNA was targeted by mir-615-5p and APOE4 protein level in APOE4＋ /-hippocampus cells was suppressed by mir-615-5p mimic transfection. Conclusion MiR-615-5p alleviates the apoptosis and degeneration of Aβ25-35-induced APOE4 ＋ /-hippocampus cells.