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目的比较辣根过氧化物酶(HRP)和荧光黄(lucifer yellow,LY)在血脑屏障模型(blood brain barrier model,BBBM)通透性测定中的使用效果。方法将Wistar乳鼠脑毛细血管内皮细胞(brain capillary endothelial cells,BCECs)和星型胶质细胞(astrocytes,As)进行共培养建立非接触BBBM;利用通透性试验进行HRP和LY对BBBM通透系数(permeability coefficient,Pe)的测定。结果建立分别与12孔和24孔板相匹配的非接触BBBM。通透性测试显示,HRP组Pe值(12孔和24孔板模型)低于LY组。HRP和LY对BBBM的Pe分别为(0.64±0.05)×10-3、(1.67±0.07)×10-3cm/min(12孔、24孔模型)和(1.08±0.16)×10-3、(3.03±0.13)×10-3cm/min(12孔、24孔模型)。结论 HRP和LY作为指示剂均能用于BBBM通透性定量测定。但LY使用简便性,安全性较高,实际中可作为优先选择。
Objective To compare the effects of horseradish peroxidase (HRP) and lucifer yellow (LY) on the permeability of blood brain barrier model (BBBM). Methods Non-contact BBBM was established by co-culturing brain capillary endothelial cells (BCECs) and astrocytes (As) in Wistar neonatal rats. Permeability test was used to study the permeability of BBBM by HRP and LY Measurement of permeability coefficient (Pe). Results Establishment of non-contact BBBM matched with 12-well and 24-well plates, respectively. Permeability tests showed that PeP values in the HRP group (12-well and 24-well plate models) were lower than in the LY group. The Pe of HRP and LY for BBBM were (0.64 ± 0.05) × 10-3, (1.67 ± 0.07) × 10-3cm / min (12 wells, 24 wells model) and (1.08 ± 0.16) × 10-3, 3.03 ± 0.13) × 10-3 cm / min (12-well, 24-well model). Conclusion Both HRP and LY can be used as quantitative indicators for BBBM permeability. But LY ease of use, higher safety, in practice, can be used as a priority.