In situ mouse carotid perfusion model:: glucose and cholesterol transport in the eye and brain
In situ mouse carotid perfusion model:: glucose and cholesterol transport in the eye and brain
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DOI：
10.1038/jcbfm.2008.34
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发表时间：
2008-08-01
影响因子：
6.3
通讯作者：
Cisternino, Salvatore
中科院分区：
文献类型：
--
作者：
Cattelotte, Julie;Andre, Pascal;Cisternino, Salvatore
采用小鼠脑原位灌注法测定血脑屏障通透性，以评估溶质在血脑屏障和血眼屏障的转运。通过颈动脉输注含氧重碳酸盐缓冲液30-120秒，用放射性标记标记物检查这一过程。使用安定估计的眼内血管流量比脑内低2.2倍。血管体积和完整性标记蔗糖和菊糖表明，2.5mL/min的灌流速度保持了这些器官的物理完整性。然而，脑血管系统的完整性对急性灌流压力比眼睛血管系统更敏感。用D-葡萄糖评价血屏障功能，其转运符合Michaelis-Menten动力学，脑的表观Km为7.6 mmo1/L，Vmax为23 lmoL/s/g，眼的表观Km为22.9 mmo1/L，Vmax为40 lmoL/s/g。通过添加ABCA1抑制剂普罗布考，胆固醇向大脑和眼睛的运输显著增强，这表明ABCA1介导的外流在小鼠脑和眼睛的血液屏障中。因此，颈动脉原位灌注适合于阐明血-脑和血-眼屏障的转运过程。
The in situ mouse brain perfusion method for measuring blood - brain barrier permeability was adapted to assess transport of solutes at the blood - brain and blood - eye barriers. The procedure was checked with radiolabeled markers in oxygenated bicarbonate- buffered fluid infused for 30 to 120 secs via a carotid artery. Vascular flow estimated with diazepam was 2.2- fold lower in the eye than in the brain. The vascular volume and the integrity markers sucrose and inulin indicated that a perfusion flow rate of 2.5 mL/ min preserved the physical integrity of these organs. However, the brain vasculature integrity was more sensitive to acute perfusion pressure than the eye vasculature. The functional capacities of blood barriers were assessed with D- glucose; its transport followed Michaelis - Menten kinetics with an apparent Km of 7.6 mmol/ L and a Vmax of 23 lmol/ sec per g in the brain, and a Km of 22.9 mmol/ L and a Vmax of 40 lmol/ sec per g in the eye. The transport of cholesterol to the brain and eye was significantly enhanced by adding the Abca1 inhibitor probucol, suggesting an Abca1- mediated efflux at the mouse brain and eye blood barriers. Thus in situ carotid perfusion is suitable for elucidating transport processes at the blood - brain and blood - eye barriers.