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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-1mRNA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-1mRNA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-1mRNA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 Ref-1 and the apoptosis of hematoma in patients with intracerebral hemorrhage. Methods According to the time from onset to operation, 30 patients with ICH were divided into six groups (6 cases), 6-12 hours group (7 cases), 12-24 hours group (5 cases), 24-72 hours group (6 cases) And ≥72h group (6 cases). The histopathology, Ref-1, apoptotic cells, Bax, and apoptosis-inhibiting genes in brain tissue were detected by HE staining, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) Change of Bcl-x gene. Seven patients with intracerebral hemorrhage who were operated within 12 hours of onset were selected as the control group. The above indexes of some brain tissues far away from the hematoma on the surgical approach were observed and compared with the surrounding tissues of the hematoma. Results Hematoxylin-eosin staining showed that the tissue around the hematoma in the control group and <6h group was basically normal, the lesion was lighter in 6 ~ 12h group, the heavier in 12 ~ 24h group, the severe damage in 24 ~ 48h group, Immunohistochemical staining showed that the apoptotic cells and Bax protein expression gradually increased 6h after onset and peaked at 12 ~ 72h (P <0.01), then gradually decreased. The expression of Bcl-x protein increased at 12-72h after onset, but there was no statistical significance (P> 0.05). The expression of Bcl-x protein was significantly decreased after 12-72h <0 · 01), then gradually raised. Bax and Bcl-xmRNA expression was similar to immunohistochemical staining. Correlation analysis showed that there was a significant negative correlation between Ref-1mRNA expression and apoptotic cells and Bax protein and mRNA expression (P <0.01), but not with Bcl-x protein and mRNA expression. Conclusion The expression of Ref-1 mRNA may play an important role in the apoptosis of cells around the hematoma and the protection of cells after intracerebral hemorrhage.