摘要
目的：脑缺血损伤的病理生理机制异常复杂，近年来的研究表明补体系统激活参与了脑出血血肿周围组织损伤的病理生理过程，但补体系统激活在脑缺血损伤中的作用尚不清楚。本实验拟通过观察大鼠脑缺血再灌注后脑组织补体C1q和C3d蛋白表达变化与脑组织含水量、血脑屏障通透性、脑组织病理改变等的关系以及眼镜蛇毒因子耗竭补体后的影响，探讨补体系统激活在缺血性脑损害中的作用，为临床脑保护治疗探索新途径。
方法：采用健康的S-D大鼠，用线栓法制备大鼠大脑中动脉闭塞（MCAO）模型，缺血2h后再灌注。测定缺血前和缺血后不同时点补体C1q和C3d蛋白表达强度、脑梗死体积、脑组织水含量、血脑屏障通透性及缺血脑组织病理和超微结构改变等指标，同时观察眼镜蛇毒因子耗竭补体后以上各指标的变化，分析其间的关系。
结果：
1．在正常脑组织中未发现补体C1q和C3d蛋白的阳性表达。
2．脑组织缺血后补体C1q和C3d蛋白的阳性表达在脑组织缺血再灌注后6h开始出现，72h达高峰，以后逐渐降低，到120h仍有较高水平表达。
3．脑组织缺血再灌注后补体C1q和C3d蛋白主要分布在缺血区皮质和基底节区的胶质细胞、神经细胞的胞浆中。
4．缺血后补体C1q和C3d蛋白的表达强度与脑组织含水量增加、血脑屏障通透性增高成平行关系。
5．眼镜蛇毒因子能显著抑制补体C1q和C3d的表达、减少脑梗死体积、减轻脑组织含水量、降低血脑屏障的通透性，减轻脑组织病理和超微结构的损害。
结论：本研究表明脑缺血后，补体系统激活参与了其脑损害的病理生理过程，眼镜蛇毒因子清除补体可以减轻脑水肿、减小脑梗死体积，降低血脑屏障的通透性，减轻脑组织的损伤，具有脑保护作用。
Object: The pathophysilogical mechanism of cerebral ischemia injury is very complicated. Recent research demonstrate that conplement activation is invovled in the pathophysilogical process of perihematom tissue injury in cerebral hemorrhage. But the effect of complement activation in cerebral ischemia is still uncertain. The aim of the study is to investigate the relationship between the expression of complement C1q and C3d protein and the change of brain water content, the permeability of BBB, pathological alteration of brain tissue in rat after MCAO. And observe the effect of complement deplection with CVF. We try to explore the action of complement activation in cerebral ischemia reperfusion injury and explore a new way for clinical neuroprotective therapy.
Method: Along with the application of intra-luminal technology,
fishing-line (0.23mm in diameter) were used to create the focal cerebral ischemia reperfusion model in healthy Sprague-Dawley rat, extract the line after occlusion 2h. We estimate the markers such as the expression intensity of complement C1q and C3d protein, brain water content, the permeability of BBB, the alteration of pathology and ultra-structure of cerebral ischemical tissue at different time before and after MCAO, observe the change of previou markers in the group of depleting complement with CVF, and analysis the relationship between them.
Result:
1．The expression of complement C1q and complement C3d protein is not observed in normal brain tissue.
2．The positive expression of complement C1q and C3d protein appear at 6h after MCAO, reach the peak at 72h, decrease gradually after then, the expression is still higher than normal tissue at 120h.
3．Complement resides in cytoplasm of the glia cell and nerve cell in
cortical and basal ganglia of ischemic area after cerebral ischemia reperfusion injury.
4．The expression intensity of complementC1q and C3d after MCAO and the raise of brain water content and permeability of BBB after MCAO is parallel.
5．CVF can significantly inhibit the expression of complement C1q
and C3d, reduce brain infarct volume, alleviate brain water content, depress the permeability of BBB, lighten the damage of in pathology and ultra-structure of brain tissue.
Conclusion： The experimental results suggested complement activation was involved in the pathophysilogical process of cerebral damage after cerebral ischemia. CVF deplete complement can alleviate brain edema, reduce infarct volume, depress the permeability of BBB, lighten the damage of brain tissue. It processes neuroprotective strategy.
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