大鼠液压冲击脑损伤Nrf2、HO-1和NQO-1动态表达变化及意义  

Expression of Nrf2,NO-1 and NQO-1 in rat brain following fluid percussion injury

作　　者：高梦颖[1] 何永昌[2] 孙国柱[2] 徐伟[2] 武利伟 

机构地区：[1]河北医科大学第二医院急诊科,石家庄050000 [2]河北医科大学第二医院神经外科,石家庄050000

出　　处：《第三军医大学学报》2016年第18期2023-2028,共6页Journal of Third Military Medical University

基　　金：河北省自然科学基金(C2008001084;H2014206383);河北省高层次人才资助项目(A201401041);河北省卫生厅资助课题(20120073;20130178)~~

摘　　要：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 alternation of nuclear factor erythroid 2-related factor 2 （Nrf2） , hemeoxygenase-1 （ HO-1 ） and NADPH ： quinine oxidoreductase （ NQO-1 ） in rats following fluid percussion injury. Methods Traumatic brain injury （TBI） model in rats was established by fluid percussion injury （n =56）. At 1, 6, 12 and 24 h, and 3 and 7 d after operation, the water content in brain tissues was quantified by dry-wet measurement, and the protein expression levels of Nrf2 （cytoplasm and nuclear）, NO-I and NQO-1 were detected by Western blotting. Also, the mRNA expression of Nil2 was measured by real-time （RT） PCR. Results Comparing with normal control （NC） group, the water content in TBI group increased after 6 h, reaching the peak at 24 h and retaining for 3 d, and then dropped （P 〈 0.05 ）. This was in agreement with the edema tendency in injured brain tissue observed by pathohistological staining. Moreover, Western blotting result showed the same change trend in protein expression of Nrf2 in nuclei, NO-1 and NQO-1 but not Nrf2 in cytoplasm. Only after 7 d, their expression was remarkably decreased but still higher than NC group （ P 〈 0.05 ）. The expression of Nrf2 in cytoplasm was consistently decreased after 12h of the injury. In addition, RT-PCR analysis showed that there were no significant differences in Nrf2 mRNA levels in TBI group at different time points （P 〉 0. 05 ）. Conclusion Nrf2 is translocated and activated in rats following fluid percussion injury, indicating that Nrf2-ARE signaling pathway may be involved in neuro-orotection via unrezulatingexpression of NO-1 and NQO-1 proteins in TBI.

关 键 词：颅脑液压冲击伤 核因子2相关因子2 血红素氧化酶-1 磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶-1 

分 类 号：R332[医药卫生—人体生理学] R642[医药卫生—基础医学]