缺氧对稳定表达人淀粉样前体蛋白HEK293细胞的存活及阿尔茨海默病相关蛋白表达的影响

Effects of hypoxia on the survival and Alzheimer's disease-related protein expression in HEK293 cells stably expression APP695 protein

目的:探讨缺氧对稳定表达人淀粉样前体蛋白的HEK293细胞(HEK293-APP695)存活及相关蛋白表达的影响,为深入研究缺氧对阿尔茨海默病的调节作用提供稳定的细胞模型。方法:利用缺氧手套箱(0.3%O2)处理HEK293-APP695细胞,CCK-8法检测细胞的存活情况;Western blot检测缺氧条件下阿尔茨海默病(AD)相关蛋白APP、APP-CTFs和BACE1的表达变化。结果:缺氧处理后,HEK293-APP695细胞的存活率明显下降,APP表达降低,其剪切体APP-CTFs表达升高。结论:缺氧导致APP剪切的增多,抑制细胞的存活,提示缺氧可能通过影响BACE1的活性在AD的发病进程中起重要的调节作用。

Objective： To further study the regulation of hypoxia on Alzheimer＇s disease （AD） pathogenesis, we investigate the effect of hypoxia on the effect of cell survival and expression of related proteins in HEK293 cells stably expressing APP695 Swedish mutantK595N/M596L （HEK293-APP695 cells）. Methods： HEK293-APP695 cells were cultured at hypoxia condition （0.3% 02）. The survival rate of HEK293- APP695 cells was measured by CCK-8 assay. The protein expression levels of APP, APP-CTFs and BACE1 were detected by Western blot. Results： The survival of HEK293-APP695 cells was obviously decreased after exposed to hypoxia. The expression of APP was reduced, and the expression of APP-CTFs was increased under hypoxia. Conclusion： Our data indicate that hypoxia accelerated cell death of HEK293- APP695 cells by increasing the cleavage of APP and production of β-secretase （β-site amyloid precursor protein cleavage enzymel, BACE1 ）.