Abstract
Purpose. This study was carried out to examine the blood-brain barrier(BBB) transport of human basic fibroblast growth factor (bFGF) andinvestigate its mechanism.
Methods. The BBB transport of 125I-bFGF was measured by severalin vivo methods including intravenous administration, in situ internalcarotid artery perfusion, and intracerebral microinjection. The in vitrobinding of 125I-bFGF was characterized using freshly prepared bovinebrain capillaries.
Results. The distribution volume of 125I-bFGF in the postvascularsupernatant increased with the perfusion time, and exceeded the spaceoccupied by the brain microvasculature and its trichloroacetic acid(TCA) precipitability was more than 90%. 125I-bFGF avidly bound toisolated bovine brain capillaries with a Bmax of 206 ± 48 pmol/mgprotein, and a Kd of 36.5 ± 15.7 nM. This binding was significantlyinhibited by unlabeled bFGF and heparin in a concentration-dependentmanner. The cationic peptides, protamine and poly-L-lysine (each 300μM), produced over 85% inhibition of 125I-bFGF binding to braincapillaries. Furthermore, glycosaminoglycans with a sulfate residue,chondroitin sulfate B and C (each 10 μg/mL) also inhibited the bindingof 125I-bFGF. The in vivo transcytosis of 125I-bFGF from the luminalside to the brain was also inhibited by the presence of heparin (10μg/mL) and poly-L-lysine (300 μM), whereas neither hyaruronic acid (10μg/mL) nor insulin (10 μM) had any effect. In addition to these results,the brain efflux index method was used to confirm that the transcytosisof 125I-bFGF from brain to blood across the BBB was negligible.
Conclusions. These results suggest that 125I-bFGF is transported acrossthe BBB, possibly by an adsorptive-mediated transcytosis mechanismthat is triggered by binding to negatively charged species on the luminalmembrane surface of the brain microvasculature, such as heparansulfate proteoglycans.
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Deguchi, Y., Naito, T., Yuge, T. et al. Blood-Brain Barrier Transport of 125I-Labeled Basic Fibroblast Growth Factor. Pharm Res 17, 63–69 (2000). https://doi.org/10.1023/A:1007570509232
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DOI: https://doi.org/10.1023/A:1007570509232