脑梗塞后炎性因子与细胞凋亡动态变化的研究

The Study of Dynamic Changes of Inflammatory Cytokine and Apoptosis after Cerebral Ischemia

目的:观察局灶性脑缺血大鼠炎性因子与细胞凋亡相关指标的改变,探讨炎性反应与细胞凋亡在脑梗塞后的变化及其内在机制。方法:采用插线法致大脑中动脉栓塞(M CAO)2 h制备局灶性脑缺血大鼠模型,分别于再灌注6、12、24、48、72和168 h取血并处死动物取相应标本。检测腹动脉血液流变学动态变化;采用酶联免疫吸附法(EL ISA)和放射免疫竞争法测定脑组织白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)含量;用苏木素-伊红(HE)染色法观察脑细胞的病理形态学变化并进行细胞计数;用原位末端缺刻标记法检测细胞凋亡情况。结果:与假手术组比较,模型组缺血侧脑组织炎症细胞因子IL-1β水平于再灌注6～24 h明显增高,TNF-α含量于再灌注24～48 h显著增多(P<0.05);再灌注6～72 h脑缺血梗死区细胞数量明显减少,12～72 h凋亡细胞大量出现(P<0.01)。结论:炎症因子IL-1β与TNF-α的生成与释放在6～48 h时增多;细胞凋亡作为缺血损伤后神经元损失的主要形式之一,在24～48 h时最为严重;缺血脑区的形态学变化在缺血后24 h最为严重。

Objective：To observe the inflammatory factors and apoptosis index changes after cerebral infarction and explore the internal mechanism.Methods： With middle cerebral artery occlusion（M CAO） 2 h,focal cerebral ischemia models prepared.And respectively after 6,12,24,48,72 and 168 h reperfusion,animals were sacrificed.Enzyme-linked immunosorbent assay（ELISA） and radioimmunoassay determination of brain tissue competition interleukin-1 β（IL1 β） and tumor necrosis factor-α（TNF-α） content;Hematoxylin-eosin（HE） staining was used to reflect changes of brain cells;TUNEL was used to detect cell apoptosis.Results： Compared with sham operation group,model group,the ischemic brain tissue inflammatory cytokines IL-1 β levels was significantly higher at 6 ～24 h reperfusion,TNF-α levels was significantly increased in the 24 ～ 48 h reperfusion（P0.05）;at 6 ～ 72 h reperfusion,infarct zone cells were significantly reduced,12～72 h,there is large numbers of apoptotic cells（P0.01）.Conclusion：The inflammatory cytokines IL-1β and TNF-α generation and release in the 6～48 h;apoptotic neurons after ischemic injury as the main form of loss of one of the most serious in the 24 ～ 48 h.