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目的研究食品添加剂亚硫酸钠(Na2SO3)对β-淀粉样物质(Aβ)前体蛋白(APP)表达的影响。方法用反转录-聚合酶链反应(RT-PCR)检测人胚肾293(HEK293)中APP基因的mRNA表达水平,用蛋白印迹法(W est-ern-b lotting)检测HEK293中APP蛋白水平。结果RT-PCR检测显示,HEK293接触0.039~10 mmol/L Na2SO324 h后细胞内APP基因的3个亚型APP695、APP751、APP770的mRNA水平均未见明显改变。蛋白印迹分析发现,10mmol/L Na2SO3可明显降低细胞内APP蛋白水平,为阴性对照组的0.908倍;而较低剂量组0.039~2.5 mmol/LNa2SO3却可以明显增加细胞内APP蛋白的水平,分别达到阴性对照组的1.596,1.630,1.556和1.446倍。结论亚硫酸钠在一定剂量范围内影响HEK293细胞内APP蛋白水平,可能对老年性痴呆的发生有一定影响,但是细胞内APP蛋白水平的增加不是由该基因转录水平增加所致。
Objective To study the effect of sodium sulfite (Na2SO3) on the expression of β-amyloid precursor protein (APP). Methods The mRNA expression of APP gene in human embryonic kidney 293 (HEK293) was detected by reverse transcription-polymerase chain reaction (RT-PCR). The protein level of APP in HEK293 was detected by Western blotting (W est-ern-b lotting) . Results RT-PCR results showed that there was no significant change in the mRNA levels of APP695, APP751 and APP770 in 3 subtypes of APP gene after exposure to 0.039-10 mmol / L Na2SO4 for HEK293. Western blot analysis showed that 10 mmol / L Na2SO3 could significantly decrease the level of APP protein in cells, which was 0.908 times of that in the negative control group, while 0.039-2.5 mmol / LNa2SO3 in the lower dose group could significantly increase the levels of APP protein Negative control group 1.596,1.630,1.556 and 1.446 times. Conclusion Sodium sulfite affects the protein level of APP in HEK293 cells in a certain dose range, which may have an impact on the development of Alzheimer’s disease. However, the increase of intracellular APP protein is not caused by the increased transcription level of this gene.