脑出血患者血肿周围组织氧化还原因子-1表达与细胞凋亡的关系

The Relationship Between the Expression of Redox Effector Factor-1 and Apoptosis in Perihematoma Tissue in Patients with Intracerebral Hemorrhage

目的：研究脑出血患者血肿周围组织氧化还原因子-1（Ref-1）表达与细胞凋亡的关系。方法：根据发病到手术的时间，将30例脑出血患者分为〈6h组（6例）、6～12h组（7例）、12～24h组（5例）、24～72h组（6例）和≥72h组（6例）。应用HE染色、免疫组化染色和逆转录-聚合酶链反应（RT-PCR）分别观察血肿旁约1cm处脑组织的病理学、Ref-1、凋亡细胞、促凋亡基因Bax和抑凋亡基因Bcl-x的变化情况。选择在发病12h内手术的7例脑出血患者作为对照组，观察手术入路上远离血肿处少许脑组织的上述指标，并与血肿周围组织进行比较。结果：HE染色显示，对照组和〈6h组的血肿周围组织基本正常，6～12h组损伤较轻，12～24h组损伤较重，24～48h组损伤严重，以后逐渐好转，8d时与对照组相似；免疫组化染色显示，凋亡细胞和Bax蛋白表达在发病6h后逐渐增高，12-72h达高峰（P〈0．01），以后逐渐下降。Bcl-x蛋白表达在发病后12～72h虽有升高趋势，但无统计学意义（P〉0．05）；RT-PCR显示，Ref-1 mRNA表达在发病后12～72h期间明显下降（P〈0．01），此后逐渐上调。Bax和Bcl-xmRNA表达与免疫组化染色结果相似。相关分析显示，Ref-1 mRNA表达与凋亡细胞和Bax蛋白及mRNA表达呈显著负相关（P〈0．01），与Bcl-x蛋白及mRNA表达无相关性。结论：Ref-1 mRNA表达可能对脑出血后血肿周围组织细胞凋亡的产生和细胞的保护起重要作用。

Objective： To study the relationship between the expression of redox effector factor- 1 （Ref-1） and apoptosis in perihematoma tissue in patients with intracerebral hemorrhage （ICH）. Methods-A total of 30 patients with ICH were divided into the following groups： 〈 6 hours （n =6）, 6-12 hours （n=7）, 12-24 hours （n =5）, 24-72 hours （n =6） and ≥72 hours （n =6）. The changes of pathology, Ref-1, apoptosis cells, promoting apoptosis gene Bax, and anti-apoptosis gene Bcl-x were observed respectively in brain tissue about 1 cm beside the hematoma by using HE staning, immunohistochemistry staining and reverse transcription polymerase chain reaction （RT-PCR）. Seven patients with ICH who were operated within 12 hours of the onset were selected as controls. The above indexes of a few brain tissues far from hematoma on the operative approaches were evaluated and compared with the perihematoma tissue. Results： HE staning showed that perihematoma tissue was almost normal in the control group and 〈 6 hours group, tissue damage was mild in the 6-12 hours group, more severe in the 12-24 hours group and severe in the 24-48 hours group, then it became better, and it was similar to the control group on day 8. Immunohistochemistry staining showed that the expression of the apoptosis cells and Bax protein increased gradually 6 hours after the onset of ICH, and it reached the maximum at 12-72 hours （P 〈 0.01 ）, then it decreased gradually. Although the expression of Bcl-x had the trend of increasing at 12-72 hours after the onset of ICH, there was no statistical significance （P 〉 0.05）. RT-PCR showed that the expression of Ref-1 mRNA decreased significantly at 12-72 hours after the onset of ICH （P 〈 0.01 ）, and then it was upregulated gradually, The expressions of Bax and Bcl-x mRNA were similar to the results of immunohistochemistry, The correlation analysis showed that the expression of Ref-1 mRNA was significantly negatively correlated with the expressions of apoptosis cells, Bax protein and mRNA （P 〈0.01 ）, and it had no correlation with the expressions of Bcl-x protein and mRNA. Conclusion： The expression of Ref-1 mRNA may play an important role in the development of apoptosis and the protection of cells in perihematoma tissue after ICH.