Department of Physiology, Loma Linda University, Loma Linda, CA 92350, USA.
J Neurosci Methods. 2011 Feb 15;195(2):206-10. doi: 10.1016/j.jneumeth.2010.12.013. Epub 2010 Dec 17.
Intracerebral hemorrhage is one of the most devastating subtypes of stroke, leaving survivors with severe neurological deficits. Disruption of the blood brain barrier (BBB) following hemorrhage results in the development of vasogenic brain edema, a most life-threatening event after such events as intracerebral hemorrhage (ICH). The Evans Blue assay is a popular method for the quantification of BBB disruption. Although this method is in common use, there are several protocols of the assay in the literature which vary in the route of administration, as well as the circulation time of the stain. In this study, we compared the amounts of accumulated stain in brain tissue following intraperitoneal versus intravenous injection at 0.5, 3 and 24h of circulation time.
58 CD-1 mice were used. Animals were divided into ICH (N=42), sham groups (N=6) and naïve (N=10). ICH animals received stereotactic injection of collagenase type VII into the right basal ganglia. Sham animals received only needle trauma. Evans Blue stain was injected 24h after collagenase injection or needle trauma. The consistency of ICH produced was characterized by estimation of hematoma volume via hemoglobin assay and neurological evaluation.
The produced hematoma and neurological deficits were well comparable between different experimental groups. There was no statistically significant difference in the results of the Evans Blue assay with regard to administration route.
The amount of Evans Blue stain accumulated in the brains of mice after ICH produced by collagenase injection was independent of the stain administration route.
脑出血是中风最具破坏性的亚型之一，使幸存者留下严重的神经功能缺损。出血后血脑屏障（BBB）的破坏导致血管源性脑水肿的发展，这是脑出血（ICH）等事件后最危及生命的事件。伊文思蓝测定法是一种常用于定量 BBB 破坏的方法。尽管这种方法被广泛应用，但文献中存在几种测定方案，其给药途径以及染料的循环时间不同。在这项研究中，我们比较了在 0.5、3 和 24 小时循环时间下，腹腔内和静脉内注射后脑组织中累积染料的量。
使用了 58 只 CD-1 小鼠。动物分为 ICH（N=42）、假手术组（N=6）和未处理组（N=10）。ICH 动物接受立体定向右基底神经节注射 VII 型胶原酶。假手术动物仅接受针损伤。胶原酶注射或针损伤后 24 小时注射伊文思蓝染料。通过血红蛋白测定和神经学评估来评估血肿量，以确定 ICH 产生的一致性。
不同实验组之间产生的血肿和神经功能缺损具有很好的可比性。伊文思蓝测定法的结果与给药途径无关，没有统计学上的显著差异。
胶原酶注射引起的 ICH 后，小鼠脑内累积的伊文思蓝染料量与染料给药途径无关。
