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用线栓法制作大鼠大脑中动脉阻塞 (MCAO)模型 ,然后用特异性细胞凋亡检测方法 (TUNEL法 ) ,对局灶缺血后不同时间点的脑组织进行观察。发现MCAO后各时间点局灶缺血范围内被标记的染色阳性的凋亡细胞主要位于局灶性缺血半暗带区 ,而且随着缺血时间的延长 ,凋亡细胞数逐渐增多 (缺血 1、3、6、1 2h分别为 3.6 0± 1 .0 2、1 5 .6 0± 2 .0 6、2 2 .4 0± 1 .85、33.0 0± 2 .83/mm2 ) ,2 4h达到高峰 ( 4 6 .6 0± 3.0 1 /mm2 ) ,4 8h以后明显减少 ( 1 3.6 0±2 .4 7/mm2 ) ,72h以后 ,凋亡表现逐渐消失 ( 5 .0 0± 1 .4 1 /mm2 ) ,各时间点的凋亡细胞数目有显著差异 (P <0 .0 1 ) ;同时两两比较表明除缺血后 1h与 72h,3h与 1 2h及 4 8h外 ,其余各点之间均有显著性差异 (P <0 .0 1 )。假手术组及对侧大脑半球各时间点基本上没有凋亡细胞出现。本研究从细胞凋亡角度重新认识急性缺血性脑血管病治疗的“时间窗” ,为缺血性卒中的治疗提供依据。
The model of middle cerebral artery occlusion (MCAO) was established by the method of thread occlusion, and then the brain tissue at different time points after focal ischemia was observed by the method of specific apoptosis detection (TUNEL). It was found that apoptotic cells stained positive after focal cerebral ischemia were mainly located in focal ischemic penumbra zone at each time point after MCAO, and the number of apoptotic cells gradually increased with the extension of ischemia Blood 1,3,6,1 2h were 3.6 0 ± 1 .0 2,1 5 .6 ± 2 .0 6,2 2 .4 ± 1 .85,33.0 ± 2 .83 / mm2), 24 h reached the peak (4.6 ± 3.0 1 / mm2), and decreased significantly after 48 h (1.360 ± 2.47 / mm2). After 72 h, the apoptotic performance gradually disappeared (5.0 ± 1 .4 1 / mm2), the number of apoptotic cells at each time point was significantly different (P <0.01); at the same time a pair of two comparison showed that in addition to ischemia 1h and 72h, 3h and 1 2h and 48h, the rest There was significant difference between each point (P <0.01). Sham-operated group and the contralateral hemisphere basically no apoptotic cells at each time point. This study re-recognized the “window of time” in the treatment of acute ischemic cerebrovascular disease from the perspective of apoptosis and provided the basis for the treatment of ischemic stroke.