摘要/Abstract
摘要: 目的 ·研究微环境变化对 PC12细胞淀粉样前体蛋白 β位点裂解酶 1(β-site amyloid precursor protein cleaving enzyme 1, BACE1)表达的影响。方法 ·分别采用毒胡萝卜素( thapsigargin,Tg)、脂多糖( lipopolysaccharide,LPS)、过氧化氢( H2O2)、缺氧培养箱和低糖培养基对 PC12细胞模拟轻度内质网应激( endoplasmic reticulum stress,ERS)、炎症环境、轻度氧化应激、低氧和低糖环境,用 Western blotting和荧光定量 PCR检测 BACE1蛋白和 BACE1 mRNA的表达水平。结果 · PC12细胞用 0.13 μmol/L Tg处理 12 h或用 0.01 mg/mL LPS处理 36 h后,BACE1蛋白和 mRNA水平显著上调(均 P<0.05);而用 0.13 mmol/L和 0.25 mmol/L H2O2处理 12 h,以及低氧环境和低糖环境下, BACE1蛋白和 mRNA均未观察到显著变化(均 P>0.05)。结论 ·轻度 ERS和炎症环境可以使 PC12细胞 BACE1的 mRNA和蛋白表达上调。
关键词: 阿尔茨海默病, 微环境, PC12细胞, 淀粉样前体蛋白 &, beta, 位点裂解酶 1, &, beta, -淀粉样蛋白
Abstract:
Objective · To investigate the effect of microenvironment on the of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in PC12 cells. Methods · The PC12 cells were respectively treated with thapsigargin (Tg), lipopolysaccharide (LPS), H2O2, hypoxia incubator and hypoglycemic medium to produce mild endoplasmic reticulum stress (ERS), inflammatory environment, mild oxidative stress, hypoxia and low glucose conditions. Western blotting and fluorescence quantitative PCR were used to detect the of BACE1 protein and BACE1 mRNA. Results · After PC12 cells were treated with Tg (0.13 μmol/L) for 12 h or LPS (0.01 mg/mL) for 36 h, BACE1 protein and mRNA levels were significantly up-regulated (P<0.05). No significant changes were observed in the of BACE1 protein and mRNA of the PC12 cells treated with H2O2 (0.13 mmol/L or 0.25 mmol/L) for 12 h, under hypoxic or hypoglycemic environment (P>0.05). Conclusion · Mild ERS and inflammatory condition can up-regulate BACE1 mRNA and protein in PC12 cells.
Key words: Alzheimers disease, microenvironment, PC12 cell, β-site amyloid precursor protein cleaving enzyme 1 (BACE1), amyloid-&beta, peptide (Aβ)
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