CAJ | 학술논문

目的：在氧糖剥夺(OGD)细胞模型中，初步探讨丁基苯酞(NBP)调节β淀粉样蛋白(Aβ)代谢中关键酶淀粉样蛋白前体β位剪切酶1(BACE1)水平的可能机制。方法采用Neuro-2a/APP695细胞，建立脑缺血体外OGD细胞模型，观察NBP对自噬抑制剂(3-MA)和自噬诱导剂雷帕霉素(Rapa)的作用。以NBP 10μmol·L-1为后续实验干预浓度分组：①对照组，不做处理；②OGD组，OGD 1 h；③OGD+NBP组；④OGD+NBP+Rapa组，200 ng·L-1 Rapa预处理1 h；⑤OGD+NBP+3-MA组，5 mmol·L-13-MA预处理1 h。采用单丹染色、Western blot及电镜等方法检测BACE1、LC3蛋白、抗凋亡蛋白(Bcl-2)和Beclin1蛋白水平以观察NBP调节自噬。结果 OGD处理后，细胞中BACE1蛋白表达降低(P<0.05)。3-MA可增加BACE1蛋白表达(P<0.01)。上调自噬及NBP可降低BACE1蛋白表达(P<0.05)。此外， NBP增加Bcl-2表达而减弱应激损伤(P<0.05)。结论 NBP可下调BACE1表达，其可能通过抑制自噬过度激活和(或)抗凋亡作用而保护细胞应对应激损伤。
목적：재양당박탈(OGD)세포모형중，초보탐토정기분태(NBP)조절β정분양단백(Aβ)대사중관건매정분양단백전체β위전절매1(BACE1)수평적가능궤제。방법채용Neuro-2a/APP695세포，건립뇌결혈체외OGD세포모형，관찰NBP대자서억제제(3-MA)화자서유도제뢰파매소(Rapa)적작용。이NBP 10μmol·L-1위후속실험간예농도분조：①대조조，불주처리；②OGD조，OGD 1 h；③OGD+NBP조；④OGD+NBP+Rapa조，200 ng·L-1 Rapa예처리1 h；⑤OGD+NBP+3-MA조，5 mmol·L-13-MA예처리1 h。채용단단염색、Western blot급전경등방법검측BACE1、LC3단백、항조망단백(Bcl-2)화Beclin1단백수평이관찰NBP조절자서。결과 OGD처리후，세포중BACE1단백표체강저(P<0.05)。3-MA가증가BACE1단백표체(P<0.01)。상조자서급NBP가강저BACE1단백표체(P<0.05)。차외， NBP증가Bcl-2표체이감약응격손상(P<0.05)。결론 NBP가하조BACE1표체，기가능통과억제자서과도격활화(혹)항조망작용이보호세포응대응격손상。
Aim To investigate the possible mechanism that dl-3-n-butylph thalide (NBP) regulate the expression of BACE1, key rate-limiting enzyme in Aβmetabolism, through oxygen-glucose-deprivation (OGD) in cells in vitro. Methods The cell model of cerebral ischemia in vitro was set up through OGD in neuro-2a/APP695 cells stably over-expressing wild-type human APP695 protein. Then autophagy regulation by NBP will be observed and analyzed by the mothod of MDC staining, immunoblotting, and transmission electron microscopy technology. Results The expression of BACE1 decreased in all groups with OGD treatments while increased in the group of 3-MA in neuro-2a/APP695 cells (P<0.01). And, the level of BACE1 declined in the group of NBP pretreatment (P<0.05). Moreover, the anti-apoptoasis protein of Bcl-2 up-regulated by NBP pretreatment against the stress damage in cells exposed to OGD. Conclusion The expression of BACE1 can be down-regulated by NBP medication of which the related mechanism may be through the inhibition of excessive autophagy and (or) anti-apoptotic pathway.