Functional and Molecular Analysis of Peptide/Protein Transport System in the Blood-Brain Barrier

血脑屏障中肽/蛋白质转运系统的功能和分子分析

• 批准号: 14572161
• 负责人: DEGUCHI Yoshiharu
• 金额: $ 2.18万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572161/
• 关键词: blood-brain barrier; adsorptive-mediated transport; heparan sulfate proteoglycan; immortalized cells; peptide transport system; opioid peptides; basic fibroblast growth factor; endocytosis; 吸着介在型輸送; カチオン性ペプチド; 脳移行性; 塩基性繊維芽細胞成長因子; ペプチド; 免疫染色 /

The adsorptive-mediated transport system (AMTS) has been believed to be one of the most advantageous system for the delivery of the pep tides and protein to the brain. The objective in this research is to clarify the adsorptive-mediated transport system (AMTS) of peptides and proteins in the blood-brain barrier (BBB) at the functional and molecular levels. in 2002 study, the transport function of basic fibroblast growth factor (bFGF), a basic peptides which is relatively large molecule (18 kDa), was examined using an immortalized mouse brain capillary endothelial cells (TM-BBB4). As a result, it was found that bFGF is internalized into TM-BBB4 via a heparin-sensitive endocytosis system. In addition, it was found that perlecan, a HSPG, is expressed on the surface of TM-BBB4 cells and the brain capillary. Accordingly, it was suggested that HSPG-mediated transport system for bFGF was functioning at the BBB as an AMTS. In 2003 study, the transport function of [D-Arg^2] dermorphin analog TAPA, a small tetra peptide, was examined. TAPA was taken up by the brain via the BBB in vivo. in addition, TAPA was internalized into TM-BBB4 cells in concentration-2 temperature-, energy-dependent manner. This internalization was inhibited by cationic peptides, but not anionic peptide and heparin. These results suggest that TAPA was transported through the BBB by the AMTS, apart form the system for bFGF. In the above study, it was suggested that there area at least, two types of AMTS at the BBB.

吸附介导转运系统（adsorptive-mediated transport system，AMTS）被认为是将肽和蛋白质递送到脑中的最有利的系统之一。本研究的目的是从功能和分子水平阐明多肽和蛋白质在血脑屏障中的吸附介导转运系统。在2002年的研究中，使用永生化小鼠脑毛细血管内皮细胞（TM-BBB 4）检测了碱性成纤维细胞生长因子（bFGF）的转运功能，碱性成纤维细胞生长因子是一种相对大分子（18 kDa）的碱性肽。结果发现，bFGF通过肝素敏感的内吞系统内化到TM-BBB 4中。此外，发现串珠素（一种HSPG）在TM-BBB 4细胞表面和脑毛细血管上表达。因此，这表明HSPG介导的bFGF转运系统在BBB中作为AMTS发挥作用。在2003年的研究中，研究了[D-Arg^2]皮吗啡类似物TAPA（一种小四肽）的转运功能。TAPA在体内通过血脑屏障被脑摄取。此外，TAPA以浓度2、温度、能量依赖的方式内化到TM-BBB 4细胞中。这种内化被阳离子肽抑制，但不被阴离子肽和肝素抑制。这些结果表明，除了bFGF系统外，TAPA还通过AMTS转运通过BBB。在上述研究中，我们认为在血脑屏障上至少存在两种类型的AMTS。

项目成果

Deguchi, Y., et al.: "Internalization of basic fibroblast growth factor at the mouse blood-brain barrier involves perlecan, a heparan sulfate proteoglycan."J.Neurochem.. 83. 381-389 (2002)

Deguchi, Y., 等人：“小鼠血脑屏障处碱性成纤维细胞生长因子的内化涉及基底膜聚糖，一种硫酸乙酰肝素蛋白聚糖。”J.Neurochem.. 83. 381-389 (2002)

Deguchi, Y., et al.: "Blood-brain barrier transport of a novel μ_1-specific opioid peptide, H-Tyr-D-Arg-Phe-β-Ala-OH(TAPA)."J.Neurochem.. 84. 1154-1161 (2003)

Deguchi, Y. 等人：“新型 μ_1 特异性阿片肽 H​​-Tyr-D-Arg-Phe-β-Ala-OH(TAPA) 的血脑屏障转运。”J.Neurochem.. 84 1154-1161（2003）

Deguchi, Y., et al.: "Application of In Vivo Brain Microdialysis to the Study of Blood-Brain Barrier Transport of Drugs."Drug Metabol.Pharmacokin.. 17. 478-490 (2002)

Deguchi, Y., 等人：“体内脑微透析在药物血脑屏障转运研究中的应用。”Drug Metabol.Pharmacokin.. 17. 478-490 (2002)

Deguchi, Y.: "Application of In Vivo Brain Microdialysis to the Study of Blood Brain Barrier Transport of Drugs"Drug Metabol. Pharmacokin.. 17. 478-490 (2002)

Deguchi, Y.：“体内脑微透析在药物血脑屏障转运研究中的应用”药物代谢。

Deguchi, Y., et al.: "Blood-brain barrier transport of a novel μ_1-specific opioid peptide, H-Tyr-D-Arg-Phe-β-Ala-OH (TAPA)"J. Neurochem.. 84. 1154-1161 (2003)

Deguchi, Y. 等人：“新型 μ_1 特异性阿片肽 H​​-Tyr-D-Arg-Phe-β-Ala-OH (TAPA) 的血脑屏障转运”J. Neurochem.. 84。 1154-1161 (2003)