知识与财富的链接
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(P0.01);(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(P0.01);(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(P0.05)。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: In order to investigate the relationship between apoptosis and the expression of NF-κB and Ⅰ-κBαprotein after intracerebral hemorrhage (ICH) in rats. Methods: The ICH model was induced by injection of Ⅶ bacterial eollagenase mixed with heparin into the right caudate nucleus of the rats. The changes of cell apoptosis were observed by terminal deoxynucleotidyl transferease mediated dUTP nick end-labeling (TUNEL) ; the expression of NF-κB and Ⅰ-κBα proteins were detected by immunohistochemical SABC method. The relative optical density of NF-κB and Ⅰ-κBα proteins were measured by Western Blot. Results: ① TUNEL positive cells appeared at 12 h after ICH, it peaked at 3 d, and it maintains the high expression at 7 d (P <0.01) ; ② The number of Ⅰ-κBα and NF-κB positive cells peaked at 12 h and 3 d after ICH. At 7 d, it still higher than that of SCG and NCG (P <0. 01) ; ③ The results of Western-blot showed that the relative optical density of NF-κB and Ⅰ-κBα proteins were most highest in ICH side 3 d and 12 h respectively after ICH. The relative optical density of NF-κB and Ⅰ-κBα in ICH side were also higher than that of normal control group at 7 d (P < 0.05). Conclusion: The results indicate that apoptotic injury occurs after ICH, NF-κB and Ⅰ-κBα involved in the cell apoptosis caused by ICH.
