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目的 研究转化生长因子 (TGF) - β1 在缺氧缺血性脑损伤 (HIBD)后脑内基因转录的变化。 方法 制备左脑 HIBD的新生大鼠模型。利用反转录聚合酶链反应 (RT- PCR)和 DNA- RNA斑点杂交技术检测新生大鼠 H IBD后不同时间海马、皮质 TGF-β1 基因转录水平的表达 ,经凝胶成像及分析系统扫描 RT- PCR扩增产物 ,以内参照甘油醛 - 3-磷酸脱氢酶 (GAPDH)半定量分析 TGF- β1m RNA的动态变化。 结果 (1) RT- PCR和斑点杂交技术最低可分别检出 10 0 fg、lpg总 RNA。 (2 )HIBD后 48h TGF- β1 m RNA出现 ,72 h为表达高峰 ,5 d后基本消失 ,高峰表达时左脑 TGF- β1 m RNA水平为右脑 2 .7倍 (P<0 .0 1) ,海马表达略高于皮质。 结论 TGF-β1 转录水平表达在 HIBD后稍晚期出现 ,其作用可能与损伤修复有关。
Objective To investigate the gene transcription of transforming growth factor (TGF) -1 in the brain after hypoxic-ischemic brain damage (HIBD). Methods The neonatal rat model of left HIBD was prepared. The expression of TGF-β1 in hippocampus and cortex at different time points after HIBD in neonatal rats was detected by reverse transcription-polymerase chain reaction (RT-PCR) and DNA-RNA dot blot hybridization. The expression of TGF-β1 was detected by gel imaging and analysis system - PCR amplification products, semi-quantitative analysis of the dynamic changes of TGF-β1m RNA by glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Results (1) RT-PCR and dot blot hybridization could detect 10 0 fg and 1pg total RNA respectively. (2) At 48h after HIBD, the expression of TGF-β1 m RNA appeared at 72 h, and disappeared after 5 d. The peak expression of TGF-β1 m RNA in left brain was 2.7 times higher than that of right brain (P <0.01) ), Hippocampus slightly higher expression of the cortex. Conclusion The expression of TGF-β1 at the later stage of HIBD may be related to the injury repair.