Lafuente J V, Cervós-Navarro J, Gutierrez Argandoña E
Department of Neuroscience, B. C. U., Leioa, Spain.
Acta Neurochir Suppl (Wien). 1994;60:139-41. doi: 10.1007/978-3-7091-9334-1_37.
Autologous serum proteins have proved to be suitable tracer to evaluate vascular permeability. The dynamic behaviour of anti-HRP immunoglobulins was studied in ultraviolet (UV) irradiation induced brain edema. Cerebral cortex of 36 anaesthetized adult rats was irradiated following a 2 x 2 mm parietal craniotomy. Immunization was carried out by 3 subcutaneous injections of 10 mg HRP in 0.5 ml complete freund adjuvant (CFA), 6, 4 and 2 weeks before the injury. Control animals were immunized only with CFA; further control animals were operated and irradiated without any previous immunization. After survival times ranging from 30 min to 24 hours, postoperation animals were transcardially perfused with 4% fresh paraformaldehyde solution in phosphate buffered saline. After postfixation at 4 degrees C, 20 microns vibratome sections were prepared for incubation with a solution of 0.05% HRP, washed and developed by the DAB reaction. The reactions showed a remarkable exsudation and spreading of anti-HRP antibodies in the edematous brain. The antigen-antibody reaction was conspicuous in animals with shorter survival periods in the necrotic area and near the lesion (1-2 mm). After a longer survival time extravasation involved the whole hemisphere. In animals with the longest survival period labeled serum proteins were found even in the white matter of the hemisphere contralateral to the injury. Endogenous tracer of BBB function is useful to study the spreading of brain edema in a delayed time after the edematous lesion.
自体血清蛋白已被证明是评估血管通透性的合适示踪剂。研究了抗辣根过氧化物酶(HRP)免疫球蛋白在紫外线(UV)照射诱导的脑水肿中的动态行为。对36只麻醉的成年大鼠,在顶骨开颅2×2毫米后对其大脑皮层进行照射。在损伤前6周、4周和2周,通过皮下注射3次10毫克HRP于0.5毫升完全弗氏佐剂(CFA)中进行免疫。对照动物仅用CFA免疫;进一步的对照动物进行手术和照射,但之前未进行任何免疫。在存活时间从30分钟到24小时不等后,对术后动物经心脏灌注含4%新鲜多聚甲醛的磷酸盐缓冲盐水溶液。在4℃后固定后,制备20微米的振动切片,用0.05% HRP溶液孵育,洗涤并通过二氨基联苯胺(DAB)反应显色。反应显示抗HRP抗体在水肿脑中显著渗出和扩散。在存活期较短的动物中,抗原 - 抗体反应在坏死区域和病变附近(1 - 2毫米)明显。存活时间较长后,渗出累及整个半球。在存活期最长的动物中,甚至在损伤对侧半球的白质中也发现了标记的血清蛋白。血脑屏障(BBB)功能的内源性示踪剂有助于研究水肿病变后延迟时间内脑水肿的扩散情况。