SorLA对阿尔茨海默病β淀粉样蛋白表达水平的影响-生物化学与分子生物学专业论文.docxVIP

PAGE PAGE 1 摘 要 目的 研究分拣蛋白相关受体 A 基因 (Sortilin-related receptor A gene, SorLA) 对阿尔茨海默病 (Alzheimer’s Disease, AD) β 淀粉样蛋白 (Amyloid β-Protein, Aβ) 表 达水平的影响，揭示 SorLA 基因与 Aβ 的表达水平之间的作用关系。 方法 培养 HEK-293(Human embryonic kidney cells) 细胞，分别将 SorLA-cDNA、SorLA-siRNA 重组质粒以 Lipofectamine? 2000 转染 HEK-293 细胞并以 G418 筛选稳定表达的细 胞，采用 Western-blot 技术检测转染组细 胞中 SorLA 蛋 白 及 淀 粉 样 前 体 蛋 白 (Amyloid precursor protein, APP) 的表达水平，采用 ELISA 技术检测转染组细胞中 Aβ 的生成量。结果 (1) 超表达 SorLA 基因转染组细胞中 APP 表达水平降低显著， Aβ 表达量降低显著(36.8±19.1)％(p0.05)；(2) SorLA-siRNA 干扰该基因的表达，转 染组细胞中 Aβ 表达量上升显著(68.2±11.6)％(p0.05)。 结论 超表达 SorLA 基因 可显著降低 Aβ 的表达量，干扰 SorLA 基因表达可显著增加 Aβ 的生成量，揭示： SorLA 基因对引发阿尔茨海默病的 Aβ 表达量具有调控作用，两者呈负相关关系， 此结论为进一步研究 Aβ 的调节机制奠定基础，为探索 AD 的治疗方法提供新思路。 关键词：SorLA；超表达；RNA 干扰；β 淀粉样蛋白；阿尔茨海默病 Effect of SorLA Gene on the Expression of β-amyloid in Alzheimer’s Disease Abstract Objective To investigate the effect of sortilin-related receptor A gene(SorLA) on the expression of amyloid β-protein(Aβ). Revealed the relation between SorLA gene and the total amount of Aβ. Methods Cultured the human embryonic kidney cells(HEK-293).Constructed the SorLA-cDNA 、 SorLA-siRNA and transfected to HEK-293T cells in mediation of Lipofectamine? 2000. Screened stability cell lines by G418 gradient concentration.The expression levels of SorLA protein and Amyloid precursor protein(APP) were analysised by Western-blot, and the total amount of Aβ were measured by ELISA. Results (1)Western-blotting showed that compared to the control group the expression levels of APP in the transfected group was reduced, and ELISA showed that the total amount of Aβ were reduced by(36.8±19.1)％(p0.05) obviously. (2)SorLA-siRNA can inhibit the expression levels of SorLA effectively，the total amount of Aβ were rosed by(68.2±11.6) ％ (p0.05). Conclusions The overexpression of SorLA gene can reduce the production of Aβ. SorLA -siRNA can inhibit the expression levels of Sor