CAJ | 학술논문

目的：明确 OGD 损伤诱导的 PC12细胞激活素 A（Activin A，ActA）信号在细胞内基于 Smad4的转导模式及作用。方法使用鼠神经生长因子（NGF，50 ng／ml）诱导大鼠嗜铬细胞瘤 PC12细胞向神经元样转化，利用无糖DMEM 培养液联合连二亚硫酸钠（1 mmol／L）构建神经元氧糖剥夺（oxygen-glucose deprivation，OGD）模型，体外模拟神经元缺血性损伤。OGD 0、3 h、6 h、12 h 后 Realtime-PCR 检测 ActA mRNA 表达水平；激光共聚焦观察免疫荧光染色后的 Smad4质核分布情况。结果NGF 诱导后 PC12细胞贴壁并伸出较长的突触，向神经元样转化。细胞 OGD处理3 h 后 ActA mRNA 表达水平较0 h 显著升高，6 h 开始下降；OGD3 h 后 Smad4在细胞核内的分布较 OGD 0 h明显增加，6小时开始下降。结论OGD 损伤可诱导神经元样 PC12细胞 ActA 信号激活，Smad4可通过质核移位介导 ActA 信号在受体水平下游的信号转导，且随时间延长呈动态变化。
목적：명학 OGD 손상유도적 PC12세포격활소 A（Activin A，ActA）신호재세포내기우 Smad4적전도모식급작용。방법사용서신경생장인자（NGF，50 ng／ml）유도대서기락세포류 PC12세포향신경원양전화，이용무당DMEM 배양액연합련이아류산납（1 mmol／L）구건신경원양당박탈（oxygen-glucose deprivation，OGD）모형，체외모의신경원결혈성손상。OGD 0、3 h、6 h、12 h 후 Realtime-PCR 검측 ActA mRNA 표체수평；격광공취초관찰면역형광염색후적 Smad4질핵분포정황。결과NGF 유도후 PC12세포첩벽병신출교장적돌촉，향신경원양전화。세포 OGD처리3 h 후 ActA mRNA 표체수평교0 h 현저승고，6 h 개시하강；OGD3 h 후 Smad4재세포핵내적분포교 OGD 0 h명현증가，6소시개시하강。결론OGD 손상가유도신경원양 PC12세포 ActA 신호격활，Smad4가통과질핵이위개도 ActA 신호재수체수평하유적신호전도，차수시간연장정동태변화。
Objective To explore the transduction mechanisms and function of the ActivinA (ActA)signaling based on Smad4 in the oxygen-glucose deprivation (OGD)PC12 cell line.Methods Nerve growth factor (NGF,50ng/ml) was used in this study to induce the differentiation of rattus PC12 pheochromocytoma cells to nerve-like cells.Then the glucose-free DMEM and hyposulfite of Na2 S2 O4 (1 mmol/L)was introduced to cells to construct an OGD model in vitro.The expression of ActA mRNA in OGD 0 h/3 h/6 h/12 h group was abserved by Realtime-PCR.And after im-munofluorescence staining,the intranuclear mobility of Smad4 in OGD 0 h/3 h/6 h/12 h group was observed under the confocal laser scanning microscope at 554 nm.Results PC12 cell was successfully differentiated into nerve-like cell af-ter NGF exposure.Compared to the group of OGD 0 h,the expression of ActA mRNA in OGD 3 h group was signifi-cantly increased,while gradually decreased in OGD 6 h and 12 h group.The fluorescence intensity of Smad4 in nuclei was increased in OGD 3 h group compared to that in OGD 0 h group,and decreased in OGD 6h group.Conclusion OGD injury could activate the ActA signaling in nerve-like PC12 cell line.Smad4 could mediate the tranduction of ActA signaling through cytoplasmic-nuclear translocation.