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目的探讨大鼠液压冲击伤后水肿脑组织核因子2相关因子2(nuclear factor erythroid-2-related factor 2,Nrf2)、血红素氧化酶-1(heme oxygenase-1,HO-1)和磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1(NADPH quinineoxidoreductase-1,NQO-1)的表达变化及其意义。方法成年雄性SD大鼠56只制作液压冲击颅脑损伤模型,术后1 h、6 h、12 h、24 h、3 d、7 d用干湿质量法测定脑组织含水量,Western blot测定Nrf2胞浆、胞核蛋白以及HO-1、NQO-1蛋白;用实时荧光定量PCR测定Nrf2 mRNA表达。结果自冲击伤后6 h,脑组织含水量开始增加,24h达高峰,持续至3 d,然后开始下降(P<0.05)。组织学观察印证了脑水肿趋势。Western blot检测结果显示胞浆Nrf2蛋白于伤后12 h开始持续降低,而胞核Nrf2蛋白则于1h开始升高,24 h达到高峰,3 d时降低,7 d后明显降低,但仍高于NC组(P<0.05)。HO-1和NQO-1蛋白表达与胞核Nrf2蛋白趋势一致。实时荧光定量PCR显示TBI组术后各时间点Nrf2 mRNA表达水平无明显变化(P>0.05)。结论液压冲击伤后出现核因子Nrf2转位激活,可能通过上调HO-1、NQO-1蛋白表达发挥神经保护作用。
Objective To investigate the expression of Nrf2, heme oxygenase-1 (HO-1) and phosphamide (AM) in rats with edema after fluid percussion injury Changes of adenine dinucleotide quinone oxidoreductase - 1 (NQO-1) expression and its significance. Methods Fifty-six adult male Sprague-Dawley rats were used to make the model of head injury by hydraulic shock. The water content of brain tissue was measured by dry-wet method at 1 h, 6 h, 12 h, 24 h, 3 d and 7 d after operation. The expression of Nrf2 Cytoplasm, nuclear protein and HO-1, NQO-1 protein; Nrf2 mRNA expression was measured by real-time fluorescence quantitative PCR. Results After 6 h, the water content of brain tissue began to increase and peaked at 24 h, reaching the level of 3 d and then decreased (P <0.05). Histological observation confirmed the trend of cerebral edema. Western blot results showed that Nrf2 protein in cytoplasm began to decrease 12 h after injury, while Nrf2 protein in nuclei increased at 1 h, peaked at 24 h, decreased at 3 d, decreased significantly after 7 d, but still higher than NC group (P <0.05). HO-1 and NQO-1 protein expression and nuclear Nrf2 protein trend. Real-time PCR showed no significant change in the expression of Nrf2 mRNA at each time point after TBI (P> 0.05). Conclusion The Nrf2 translocation activated by hydraulic shock injury may play a neuroprotective role by up-regulating the expression of HO-1 and NQO-1.