CAJ | 학술논문

目的观察体外培养的大鼠海马神经元经氧糖剥夺损伤后信号转导和转录激活因子-3(signal transducer and activator of transcription factor-3,STAT3)磷酸化及其核转移,探讨体外模拟的脑缺氧、缺血损伤细胞模型中STAT3信号通路的动态变化. 方法选择新生24h内SD大鼠,取双侧海马神经元,用DMEM/F12培养基培养9d,氧糖剥夺处理4h,建立海马神经元氧糖剥夺损伤模型.运用免疫荧光技术,在激光共聚焦扫描显微镜下观察磷酸化STAT3(p-STAT3)的表达部位.采用Western bloting技术检测海马神经元氧糖剥夺后不同时相点p-STAT3表达强度的变化. 结果对照组p-STAT3在胞核表达不明显；而模型组在建模后1 h p-STAT3在胞核内即有表达,3h达到高峰,各时相点与对照组比较,差异有统计学意义(P＜0.05). 结论氧糖剥夺损伤诱导海马神经元胞核中p-STAT3水平明显升高,提示海马神经元氧糖剥夺损伤后STAT3信号转导通路被过度激活.
목적 관찰체외배양적대서해마신경원경양당박탈손상후신호전도화전록격활인자-3(signal transducer and activator of transcription factor-3,STAT3)린산화급기핵전이,탐토체외모의적뇌결양、결혈손상세포모형중STAT3신호통로적동태변화. 방법 선택신생24h내SD대서,취쌍측해마신경원,용DMEM/F12배양기배양9d,양당박탈처리4h,건립해마신경원양당박탈손상모형.운용면역형광기술,재격광공취초소묘현미경하관찰린산화STAT3(p-STAT3)적표체부위.채용Western bloting기술검측해마신경원양당박탈후불동시상점p-STAT3표체강도적변화. 결과 대조조p-STAT3재포핵표체불명현；이모형조재건모후1 h p-STAT3재포핵내즉유표체,3h체도고봉,각시상점여대조조비교,차이유통계학의의(P＜0.05). 결론 양당박탈손상유도해마신경원포핵중p-STAT3수평명현승고,제시해마신경원양당박탈손상후STAT3신호전도통로피과도격활.
Objective To observe the phosphorylation level and nuclear translocation of signal transducer and activator of transcription factor-3 (STAT3) in hippocampal neurons induced by oxygen and glucose deprivation in vitro and discuss the dynamic changes of STAT3 signal pathway in an in vitro cell model of brain hypoxia and ischemia.Methods Hippocampal neurons from newly born SD rats (within 24 hours from birth) were cultured with DMEM/F12 for nine days,and then were transferred to oxygen and glucose deprivation environment for four hours to establish experimental cell models.The distribution of phosphorylated STAT3 (p-STAT3) in the hippocampal neurons in different groups was observed under laser scanning confocal microscope after immunofluorescence staining.Expression intensity of p-STAT3 at different time points after oxygen and glucose deprivation in the hippocampal neurons was detected by Western blotting.Results Expression of p-STAT3 was unobvious in the nucleus of the control group,but it was observed in the nucleus of the model group one hour after modeling,and peaked at three hour.Expression levels of p-STAT3 in the hippocampal neurons at each time point between the two groups showed significant difference (P ＜ 0.05).Conclusion Oxygen and glucose deprivation induces noticeable up-regulation of p-STAT3 in the hippocampal neuronal nucleus,which indicates the overactivation of signal transduction pathway of STAT3.