Objective To investigate the expression of NF-κB mRNA after brain contusion in rats and the relationship between the NF-κB mRNA expression and the wound age. Methods Sixty-four Wistar rats were randomly divided into 8 groups. One control group and seven experimental groups which were wounded and then taken samples respectively in 0. 5h,6h, 12h, 1d,3d,7d and 14d after brain contusion. Total RNA was extracted from each group and the 1st strand cDNA was synthesized by reverse transcription polymerase chain reaction. The amount of NF-κB mRNA in every sample was quantified by using RPL32 intrinsic fluorescent assay and the data was compared by Deta Ct with that of the control group. Results The expression of NF-κB mRNA began to increase in 0.5h post injury,which was 9. 19 times as much as the control group;and reached the maximum level in 12h in marginal area of injury focus,which was 13.40 times of the control group;then the expression decreased gradually and sustained at a high level up to 14d ,which was 2.59 times as that of the control groups. Conclusion The time-related expression of NF-κB mRNA after brain contusion may be useful for estimation of early wound age.%目的 应用实时定量PCR技术检测大鼠脑挫伤后脑组织中NF-κB mliNA的时序性表达规律.方法 Wistar大鼠64只,随机分为1个对照组和7个实验组,每组8只.建立大鼠脑挫伤模型,于伤后0.5h、6h、12h、1d、3d、7d及14d取材,一步法提取总RNA,检测其完整性和浓度,按照0.4μg总RNA量逆转录合成第一链cDNA,以RPL32为参比基因进行荧光定量检测,采用△Ct法比较其与对照组脑组织中NF-κB mRNA表达的倍数关系.结果 NF-κB mRNA的表达于脑挫伤后呈现先增高后下降的表达规律,0.5h升高,为对照组的9.190倍,伤后12h升至最高,为对照组的13.40倍,随后开始下降,至伤后14d时仍维持较高水平,为对照组的2.585倍.结论 大鼠脑挫伤后脑组织中NF-κB mRNA的表达随着损伤时间呈现先增高后降低的规律性变化,可望作为推断早期脑挫伤的客观指标之一.
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