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目的探讨血红素结合蛋白(HPX)是否通过上调血红素氧合酶-1(HO-1)改善大鼠脑缺血再灌注(IR)损伤。方法 60只雄性SD大鼠,7~8周龄,体重250~280 g,采用随机数字表法分为五组:假手术(Sham)组、缺血再灌注(MCAO)组、溶剂(叠氮钠)(MCAO+Vehicle)组、HPX(MCAO+HPX)组和HO-1抑制剂组(MCAO+HPX+ZnppⅨ)组,每组12只。采用大脑中动脉栓塞(MCAO)法制备大鼠缺血再灌注模型,MCAO组、MCAO+Vehicle组、MCAO+HPX组、MCAO+HPX+ZnppⅨ组大鼠分别在IR即刻经侧脑室单次注射生理盐水(10μl,0.9%)、叠氮钠(10μl,0.1%)、HPX(10μl,1.86 g/L)和10μl HPX+ZnppⅨ混合液(HPX 1.86 g/L+叠氮钠0.1%)。IR后24 h、7 d分别采用Carcia评分法评价大鼠神经行为学评分(NBS),采用实时荧光定量聚合酶链式反应(RT-PCR)法检测半暗带区脑组织中HO-1 m RNA水平。结果 IR后24 h Sham组、MCAO组、MCAO+Vehicle组、MCAO+HPX组、MCAO+HPX+ZnppⅨ组大鼠的NBS评分分别为(15.50±2.43)、(5.17±1.60)、(5.00±2.10)、(10.83±1.47)、(5.33±1.97)分,比较差异有统计学意义(F=34.819,P<0.05);IR后7 d Sham组、MCAO组、MCAO+Vehicle组、MCAO+HPX组、MCAO+HPX+ZnppⅨ组大鼠的NBS评分分别为(16.83±1.60)、(8.83±3.31)、(9.17±3.19)、(12.50±3.08)、(9.33±2.07)分,比较差异有统计学意义(F=13.113,P<0.05)。与Sham组相比,MCAO组大鼠IR后24 h、7 d缺血半暗带区脑组织HO-1 m RNA水平显著增加(P<0.05);与MCAO组相比,MCAO+Vehicle组大鼠IR后24 h、7 d缺血半暗带区脑组织HO-1 m RNA水平无明显变化(P>0.05);与MCAO+Vehicle组大鼠相比,MCAO+HPX组大鼠IR后24 h、7 d缺血半暗带区脑组织HO-1 m RNA水平显著升高(P<0.05)。结论 HPX可通过上调HO-1表达改善大鼠脑缺血再灌注后神经行为学表现。
Objective To investigate whether heme-binding protein (HPX) improves cerebral ischemia-reperfusion (IR) injury in rats by upregulating heme oxygenase-1 (HO-1). Methods Sixty male Sprague Dawley rats aged 7-8 weeks and weighing 250-280 g were randomly divided into five groups: sham group, MCAO group, (MCAO + Vehicle) group, HPX (MCAO + HPX) group and HO-1 inhibitor group (MCAO + HPX + ZnppⅨ) MCAO group, MCAO + Vehicle group, MCAO + HPX group, MCAO + HPX + ZnppⅨ group were injected with IR immediately through the lateral ventricle single injection of physiology Saline (10 μl, 0.9%), sodium azide (10 μl, 0.1%), HPX (10 μl, 1.86 g / L) and 10 μl of HPX + ZnppIX cocktail (HPX 1.86 g / L + sodium azide 0.1%). The neurological behavioral score (NBS) was evaluated by Carcia score at 24 and 7 d after IR, respectively. The expression of HO-1 m in brain tissue was detected by real-time fluorescence quantitative polymerase chain reaction (RT-PCR) RNA level. Results The NBS scores of Sham group, MCAO group, MCAO + Vehicle group, MCAO + HPX group and MCAO + HPX + ZnppⅨ group were (15.50 ± 2.43), (5.17 ± 1.60), (5.00 ± 2.10 ), (10.83 ± 1.47) and (5.33 ± 1.97), respectively. The difference was statistically significant (F = 34.819, P <0.05) , And the scores of NBS in MCAO + HPX + ZnppⅨ group were (16.83 ± 1.60), (8.83 ± 3.31), (9.17 ± 3.19), (12.50 ± 3.08) and (9.33 ± 2.07) Significance (F = 13.113, P <0.05). Compared with Sham group, the level of HO-1 mRNA in brain tissue of ischemic penumbra in MCAO group was significantly increased 24 h and 7 d after IR (P <0.05). Compared with MCAO group, MCAO + Vehicle group Compared with MCAO + Vehicle group, the level of HO-1 m RNA in brain tissue of ischemic penumbra in 24 h and 7 d IR rats was not significantly changed (P> 0.05) The levels of HO-1 m RNA in brain tissue in ischemic penumbra zone on day 7 and day 7 were significantly increased (P <0.05). Conclusion HPX can improve neurological behavior after cerebral ischemia-reperfusion in rats by up-regulating the expression of HO-1.