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目的:探讨大鼠脑出血(ICH)后细胞凋亡及其与NF-κB和I-κBα蛋白表达的关系和演变规律,本实验用Ⅶ胶原酶联合肝素注入大鼠右侧脑纹状体制备脑出血模型。方法:TUNEL染色(末端脱氧核糖核苷转移酶介导的dUTP缺口末端标记法)观察细胞凋亡的变化;免疫组化SABC法检测NF-κB和I-κBα蛋白的表达;Western Blot技术检测NF-κB和I-κBα蛋白相对光密度值。结果:(1)ICH组TUNEL阳性细胞于出血后12h可检测到,3d达高峰,7d仍有较高表达(P<0.01);(2)ICH组I-κBα和NF-κB蛋白免疫阳性细胞分别于出血12h、3d达高峰,7d仍高于假手术组和正常对照组(P<0.01);(3)Western Blot结果显示NF-κB相对光密度比值在ICH3d出血侧达高峰,I-κBα蛋白相对光密度比值在ICH组ICH侧12h达高峰,7d时仍明显高于正常对照组(P<0.05)。结论:ICH后存在细胞凋亡;NF-κB和I-κBα参与了ICH所致的细胞凋亡。
Objective: To investigate the relationship between the apoptosis and the expression of NF-κB and I-κBα after intracerebral hemorrhage (ICH) in rats. In this experiment, collagenase VII and heparin were injected into the right striatum of rats Intracerebral hemorrhage model. Methods: The changes of apoptosis were observed by TUNEL staining (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling); the expression of NF-κB and I-κBα protein was detected by immunohistochemical SABC method; NF -κB and I-κBα protein relative optical density values. Results: (1) TUNEL positive cells in ICH group were detectable at 12h after hemorrhage and peaked at 3d, and remained high at 7d (P <0.01). (2) Immunohistochemical staining of I-κBα and NF- (P <0.01). (3) Western Blot results showed that the ratio of relative optical density of NF-κB peaked at the hemorrhage side of ICH3d, the expression of I-κBα The ratio of protein relative optical density peaked at 12 hours in ICH side of ICH group and was still significantly higher than that of normal control group on the 7th day (P <0.05). Conclusion: There is apoptosis after ICH. NF-κB and I-κBα are involved in ICH-induced apoptosis.