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目的探讨大鼠全脑缺血再灌注后皮质组织中HAX-1蛋白(HS1-associated protein X-1,HAX-1)与皮质神经元凋亡的关系。方法建立大鼠四血管法全脑缺血模型,将实验动物分为5组(n=5):正常组,缺血6、24、48、72h组。采用HE染色,观察大鼠皮层组织病理变化;采用免疫组化、TUNEL法检测大鼠全脑缺血再灌注后HAX-1蛋白的表达以及皮质神经元凋亡蛋白Caspase-3的表达情况。结果HAX-1蛋白在缺血后6h表达最高(37.60±3.45),24、48、72h降低[分别为(11.40±1.14)、(10.40±1.52)、(9.80±1.30)],且低于正常水平(P<0.01);Caspase-3蛋白随着缺血时间的延长逐渐增高[6、24、48、72h分别为(72.80±5.49)、(106.20±6.91)、(129.00±19.74)、(166.20±15.32)](P<0.01),并伴随神经元凋亡的数目逐渐增加[(92.00±9.06)、(133.80±10.64)、(157.00±10.83)、(187.80±14.96)]。结论全脑缺血再灌注后皮质神经元中HAX-1蛋白在短期缺血中表达升高,可能参与神经元的抗凋亡,随着缺血时间延长,其表达水平降低,与神经元的凋亡增强相关。
Objective To investigate the relationship between the expression of HAX-1 protein (HAX-1) and cortical neurons in cortical tissues after global cerebral ischemia-reperfusion in rats. Methods The rat model of global cerebral ischemia with four vessels was established. The experimental animals were divided into 5 groups (n = 5): normal group, ischemia group 6,24,48,72h. The pathological changes of cortex were observed by HE staining. The expression of HAX-1 protein and the expression of Caspase-3 in cortical neurons were detected by immunohistochemistry and TUNEL after global cerebral ischemia and reperfusion. Results The expression of HAX-1 protein was the highest at 6h after ischemia (37.60 ± 3.45) and decreased at 24, 48 and 72h (11.40 ± 1.14, 10.40 ± 1.52 and 9.80 ± 1.30, respectively) (72.80 ± 5.49), (106.20 ± 6.91), (129.00 ± 19.74), (166.20, P <0.01). Caspase-3 protein gradually increased with the prolongation of ischemia time ± 15.32)] (P <0.01), and the number of apoptotic neurons increased gradually [(92.00 ± 9.06), (133.80 ± 10.64), (157.00 ± 10.83), (187.80 ± 14.96)]. Conclusion The expression of HAX-1 protein in cortical neurons after global cerebral ischemia-reperfusion is increased in short-term ischemia, which may be involved in the anti-apoptosis of neurons. With the prolongation of ischemia, the expression of HAX- Apoptosis related.