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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 mRNA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、12h1、d、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。
Objective To detect the temporal pattern of NF-κB mRNA expression in rat brain after contusion by real-time quantitative PCR. Methods Sixty-four Wistar rats were randomly divided into 1 control group and 7 experimental groups, 8 in each. The rat model of contusion was established and the total RNA was extracted at 0.5h, 6h, 12h1, d, 3d, 7d and 14d after injury. The total RNA was extracted by one-step method. The integrity and concentration of 0.4μg total RNA were reverse transcribed Chain cDNA, RPL32 as a reference gene for quantitative detection of fluorescence, using △ Ct method compared with the control group, NF-κB mRNA expression in the multiple relationship. Results The expression of NF-κB mRNA increased first and then decreased after cerebral contusion, increased 0.5h, which was 9.190 times that of the control group. The expression of NF-κB mRNA increased to 13.40 times of the control group at 12h after injury, 14d after injury to maintain a high level of 2.585 times the control group. Conclusion The expression of NF-κB mRNA in brain tissue of rats with brain contusion increases first and then decreases with the time of injury, which is expected to be one of the objective indexes to infer the early cerebral contusion.