Establishment of an In Vitro Model of "Blood Brain Barrier" and an Attempt with it to Develop a Brain-Specific Transport Vector

“血脑屏障”体外模型的建立及脑特异性转运载体的尝试

• 批准号: 04558024
• 负责人: YAMAMOTO Hiroshi
• 金额: $ 11.26万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04558024/
• 关键词: blood brain barrier; vascular endothelial cells; pericytes; astroglial cells; advanced glycation endproducts; diabetic microangiopathy; aldosterone; vascular endothelial growth factor; トランス分化; 血管平滑筋細胞; ミネラルコルチコイドレセプター; vascular endothelial grow

(1) In the present study, we have established an in vitro model of "blood brain barrier". With vascular endothelial cell, pericyte and astroglial cell co-culture systems, the barrier as well as transport activities selective to brain vessels were reconstituted, which passed polycationic albumin but excluded native albumin or inulin in the physiological polarity. This model would seem to be useful in screeeing substances that could act on the central nervous system and in evaluating their delivery into the brain. The present study has also demonstrated that astroglial cells are capable of "transdifferentiating "non-brain-type endothelial cells into brain-type ones : human umbilical vein endothelial cells co-cultured with astroglial cells did express the brain endothelium-specific genes, including those for gamma-transglutamyl transpeptidase, transferrin receptor and P-glycoprotein, and aquired the anti-inulin barrier propert. Although we failed to develop brain-specific transport vectors, new discoveries (2) that advanced glycation endproducts selectively injure pericytes through interactions with their cell-surface receptors, (3) that endothelial cells and vascular smooth muscle cells possess the devices both for synthesizing and for responding to aldosterone, a steroid implicated in the regulation of blood pressure, and (4) that hypoxia-induced proliferation of vascular cells in mediated by autocrine vascular endothelial growth factor, were made during the course of this study, and we have thus proposed novel mechanisms underlying the development and progression of various human vascular disorders including diabetic microangiopathy, hypertension and angiogenesis.

(1)在本研究中，我们建立了一个“血脑屏障”的体外模型。在血管内皮细胞、周细胞和星形胶质细胞共培养系统中，重建了对脑血管的屏障和选择性转运活动，其通过聚阳离子白蛋白，但排除生理极性的天然白蛋白或菊粉。这个模型似乎在筛选作用于中枢神经系统的物质和评估它们进入大脑的过程中很有用。本研究还证实星形胶质细胞具有将非脑型内皮细胞“转分化“为脑型内皮细胞的能力：与星形胶质细胞共培养的人脐静脉内皮细胞表达脑内皮特异性基因，包括γ-转谷氨酰转肽酶、转铁蛋白受体和P-糖蛋白基因，并获得抗菊粉屏障的特性。虽然我们未能开发出脑特异性转运载体，但新发现（2）晚期糖基化终产物通过与细胞表面受体的相互作用选择性地损伤周细胞，（3）内皮细胞和血管平滑肌细胞具有合成和响应醛固酮（一种与血压调节有关的类固醇）的装置，（4）缺氧诱导的血管细胞增殖是由自分泌的血管内皮生长因子介导的，因此我们提出了各种人类血管疾病包括糖尿病微血管病、高血压和血管生成的发生和发展的新机制。

项目成果

Yasuko Kamiya: "Mutations in genes for acetylcholinesterase intensify lethality by acrylamide in Caenorhabditis elegans." Neuroscience letters. 145. 37-39 (1992)

Yasuko Kamiya：“乙酰胆碱酯酶基因突变增强了丙烯酰胺对秀丽隐杆线虫的杀伤力。”

Koji Nata: "Structure determination and evolution of the chicken cDNA and gene encoding prepropancreatic polypeptide" Gene. 130. 183-189 (1993)

Koji Nata：“鸡 cDNA 和编码前胰多肽的基因的结构测定和进化”基因。

S.Yamagishi, M.Taniguchi, S.Harada, C.-C.Hsu., K.Ohsawa, K.Kobayashi, H.Yamamoto: "A role of AGE receptor in the development of diabetic microangiopathy" Antisense Research and Development. (in press). (1995)

S.Yamagishi、M.Taniguchi、S.Harada、C.-C.Hsu.、K.Ohsawa、K.Kobayashi、H.Yamamoto：“AGE 受体在糖尿病微血管病发展中的作用”反义研究与开发。

Motohiro Nomura and Hiroshi Yamamoto: "Hypoxia and Angiogenesis(in Japanese)" in "Vascular Remodeling and Its Pathogenesis" , Saishin-Igaku Vol 50 (6) , 1995, Saishin-Igaku-Sha, Osaka.

Motohiro Nomura 和 Hiroshi Yamamoto：“血管重塑及其发病机制”中的“缺氧和血管生成”，Saishin-Igaku Vol 50 (6)，1995，Saishin-Igaku-Sha，大阪。

Haruhiko Hatakeyama: "Vascular Aldosterone:Biosynthesis and a Link to Angiotensin II-Induced Hypertophy of Vascular Smooth Muscle Cells." J.Biol.Chem.269. 24316-24320 (1994)

Haruhiko Hatakeyama：“血管醛固酮：生物合成及其与血管紧张素 II 诱导的血管平滑肌细胞肥大的联系。”