脑梗塞后炎性因子与细胞凋亡动态变化的研究

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。

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

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.