摘要
目的:探讨STAT3,5基因在缺氧性海马神经元细胞损伤中的可能作用。方法:分离新生Wistar大鼠双侧海马进行体外培养,采用Tripure试剂提取海马神经元细胞的总RNA,自行设计大鼠STAT3,STAT5和阳性对照HPRT的PCR引物序列,通过半定量的逆转录多聚酶链反应(RT-PCR)技术检测常氧组、低氧2,6,12h组海马神经元细胞的STAT3和STAT5基因的表达水平;采用PCR产物纯化试剂盒纯化PCR产物,而后用四色荧光法直接测序。结果:图像分析结果显示低氧培养2h组海马神经元细胞中STAT3,5mRNA表达水平升高,目的片段与HPRT的光密度比值分别为0.57±0.11和0.64±0.09,在低氧6h组(0.85±0.14和0.86±0.11)达最高,与正常组(0.34±0.12和0.40±0.08)比较差别均有显著性意义(t=-24.59～-7.37,P<0.01)。低氧12h组的表达量低于6h组,但高于正常组(P<0.01)。测序显示,PCR扩增的产物STAT3,STAT5与大鼠Genbank中的序列完全一致。结论:低氧增强海马神经元细胞中STAT3,5基因的表达,在低氧所致的脑损伤中可能发挥重要作用。
AIM:To explore the effect of signal transducers and activators of transcription(STAT) on the injury of hypoxia hippocampal neurons. METHODS:Bilateral hippocampal cells of newborn rats were separated for culture in vitro.The total ribonucleic acid(RNA) of hippocampal neurons was extracted by using Tripure reagen,the PCR primer ranking of STAT3,STAT5 and positive control HPRT were designed.The gene expression level of STAT3 and STAT5 in hippocampal neurons were detected by semi quantitative reverse transcriptase and polymerase chain reaction(RT PCR) in common oxygen group(control group),2 h under hypoxia group(group 1)6 h under hypoxia group(group 2),12 h under hypoxia group(group 3).The products of PCR were purified by the purification kits,and were sequenced directly by the fluorescence method of four colours. RESULTS:The analysis results of image showed that the expression level of STAT3 messenge RNA(mRNA) and STAT5 mRNA were increased in the hippocampal neurons in group 1(0.57±0.11,0.64±0.09),the ratios of objective fragment and HPRT optical density were 0.57±0.11,0.64±0.09,it reached the highest in group 2(0.85±0.14, 0.86±0.11),it was significantly different from that in the control group(0.34±0.12,0.40±0.08)(t=-24.59--7.37,P< 0.01).The expression in group 3 was lower than that in group 2,but higher than that in the control group(P< 0.01).The sequencing showed that the amplificated product of PCR STAT3 and STAT5 were completely the same to that in the Genbank of rats. CONCLUSION:The gene expression of STAT3 and 5 in the hippocampal neurons reinforced by hypoxia plays an important role in brain injury caused by hypoxia.
出处
《中国临床康复》
CSCD
2004年第1期66-67,共2页
Chinese Journal of Clinical Rehabilitation
