CAJ | 학술논문

目的：观察低氧预处理（ HPC）对氧糖剥夺（ OGD）损伤的PC12细胞的作用，同时探讨自噬在其中的作用。方法：培养的PC12细胞按照处理因素分为6组：对照组、HPC模型组、3－甲基腺嘌呤（3－MA）组、低氧预处理后氧糖剥夺（HPC＋OGD）组、3－甲基腺嘌呤预处理后行低氧预处理及氧糖剥夺（3－MA＋HPC＋OGD）组和OGD组。 CCK－8检测细胞存活率；caspase－3活性检测试剂盒检测酶活性；TUNEL染色和流式细胞术检测细胞凋亡的情况；Western blot法检测自噬相关蛋白LC3、beclin－1和凋亡相关蛋白caspase－3的表达。结果：与对照组相比，OGD组细胞存活率明显降低；与3－MA＋HPC＋OGD组及OGD组相比，HPC＋OGD组细胞存活率明显升高（ P＜0．05）。与对照组相比，OGD组细胞的caspase－3酶活性明显升高；与3－MA＋HPC＋OGD组及OGD组相比，HPC＋OGD组的caspase－3酶活性明显降低（ P＜0．05）。与对照组相比OGD组细胞凋亡明显增多；HPC＋OGD组与OGD组相比凋亡明显减少（P＜0．05）。此外，与对照组相比，OGD组的激活型caspase－3蛋白水平明显升高（P＜0．05）；且HPC＋OGD组与OGD组相比激活型caspase－3蛋白水平明显减少而LC3、beclin－1的蛋白水平明显升高（ P＜0．05）。结论：HPC抗OGD损伤的机制可能与其激活细胞自噬有关。
목적：관찰저양예처리（ HPC）대양당박탈（ OGD）손상적PC12세포적작용，동시탐토자서재기중적작용。방법：배양적PC12세포안조처리인소분위6조：대조조、HPC모형조、3－갑기선표령（3－MA）조、저양예처리후양당박탈（HPC＋OGD）조、3－갑기선표령예처리후행저양예처리급양당박탈（3－MA＋HPC＋OGD）조화OGD조。 CCK－8검측세포존활솔；caspase－3활성검측시제합검측매활성；TUNEL염색화류식세포술검측세포조망적정황；Western blot법검측자서상관단백LC3、beclin－1화조망상관단백caspase－3적표체。결과：여대조조상비，OGD조세포존활솔명현강저；여3－MA＋HPC＋OGD조급OGD조상비，HPC＋OGD조세포존활솔명현승고（ P＜0．05）。여대조조상비，OGD조세포적caspase－3매활성명현승고；여3－MA＋HPC＋OGD조급OGD조상비，HPC＋OGD조적caspase－3매활성명현강저（ P＜0．05）。여대조조상비OGD조세포조망명현증다；HPC＋OGD조여OGD조상비조망명현감소（P＜0．05）。차외，여대조조상비，OGD조적격활형caspase－3단백수평명현승고（P＜0．05）；차HPC＋OGD조여OGD조상비격활형caspase－3단백수평명현감소이LC3、beclin－1적단백수평명현승고（ P＜0．05）。결론：HPC항OGD손상적궤제가능여기격활세포자서유관。
AIM: To examined the effects of hypoxic preconditioning ( HPC) on oxygen-glucose deprivation ( OGD)-induced PC12 cells, and to investigate its possible mechanisms of autophagy .METHODS: Cultured PC12 cells were randomly divided into control group , HPC group, 3-methyladenine (3-MA) group, HPC+OGD group, 3-MA+HPC+OGD group and OGD group .CCK-8 assay was used to detect the cell viability .The caspase-3 activity was also tested . TUNEL staining and flow cytometry were used to detect the cell apoptosis .The protein levels of apoptosis-related protein caspase-3 and autophagy-marked protein LC3-2 and beclin-1 were determined by Western blot .RESULTS:Compared with control group, the viability of PC12 cells was significantly reduced , and the activity of caspase-3 was significantly increased in OGD group.Compared with 3-MA+HPC+OGD group and OGD group , the viability of PC12 cells was significantly in-creased, and the activity of caspase-3 was significantly reduced in HPC +OGD group (P<0.05).The PC12 cell injury was apparent after OGD with a great increase in the apoptotic rate (P<0.05).Compared with OGD group, the apoptotic rate significantly decreased in HPC +OGD group ( P<0.05 ) .Compared with control group , the protein level of cleaved caspase-3 was significantly increased in OGD group ( P<0.05) .Compared with OGD group , the protein level of cleaved caspase-3 was significantly decreased , and the levels of LC3-2 and beclin-1 were significantly increased in HPC +OGD group (P<0.05).CONCLUSION:OGD decreases cell survival and induces apoptosis .Activation of cell autophagy may be the mechanism by which hypoxic preconditioning protects the PC 12 cells from OGD induced injury .