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Determination of the blood–brain barrier (BBB) permeability

ML Min Liu
ZX Zhipeng Xu
LW Long Wang
LZ Lixia Zhang
YL Yi Liu
JC Jiangbei Cao
QF Qiang Fu
YL Yanhong Liu
HL Hao Li
JL Jingsheng Lou
WH Wugang Hou
WM Weidong Mi
YM Yulong Ma
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Immediately after reperfusion, 4 ml/kg of 2% Evans blue (Sigma Aldrich) was injected into the right jugular vein (n = 6). The animals were euthanatized 24 h after reperfusion. For Evans blue extravasation assay, coronal sections were cut into 2-mm slices and were photographed. The blue-stained area was defined as BBB permeable. For quantitative measurements, the ischemic hemispheres were homogenized in N, N-dimethylformamide (10 ml/kg, Sigma Aldrich), incubated for 18 h at 55 °C, and centrifuged at 3000 rpm for 10 min [22]. The supernatants were analyzed at 620 nm by spectrophotometry. The EB content was calculated from its standard curve as a measure of the BBB permeability change.

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