1994 Fiscal Year Final Research Report Summary
Establishment and application of in vitro Blood-Brain-Barrier experimental system with cultured brain capillary endothelial cells and astrocytes.
| Project/Area Number |
04670863
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
NAGASHIMA Tatsuya Kobe University School of Medicine, Department of Neurosurgery, Assistant professor, 医学部, 助手 (80201680)
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| Co-Investigator(Kenkyū-buntansha) |
KOKUNAI Takashi Kobe University School of Medicine, Department of Neurosurgery, Assistant Profes, 医学部・附属病院, 講師 (30178248)
富田 洋司 神戸大学, 医学部・附属病院, 助手 (90252775)
TAMAKI Norihiko Kobe University School of Medicine, Department of Neurosurgery, Professor, 医学部, 教授 (10030941)
TOMITA Youji Kobe University School of Medicine, Department of Neurosurgery, Assistant Profes
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| Project Period (FY) |
1992 – 1994
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| Keywords | BLOOD-BRAIN BARRIER / ENDOTHELIAL CELLS / BRAIN / ASTROCYTES / CEREBRAL METABOLISM / ATRIAL NATRIURETIC PEPTIDES |
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| Research Abstract |
(1) IN VITRO BBB EXPERIMENTAL SYSTEM WAS ESTABLISHED.
Brain capillary endothelial cells were isolated from rat brain cortices by modified Audus' enzymatic method. The cells were cultures on a polycarbonate mesh to study transport of 14C-sucrose and electric resistance. The membrane showed increased electrical resistance and barrier to the C14-sucrose transport. Coculture with astrocytes obtained from the newborn rat cortex induced higher electrical resistance and higher barrier function to 14C-sucrose movement.
(2) THERE IS NO ENZYMATIC BARRIER TO LEUKOTRIENE C4 ON CULTURED BRAIN CAPPILARY ENDOTHELIAL CELLS.
The cultured endothelial cells contained very low concentration of gamma-GTP.The application of leukotriene C4 on the endothelial cell monolayr did not increase the transport of 14C-Sucrose. It indicates that gamma-GTP does not work as a enzymatic barrier of leukotriene C4.
(3) ANOXIC ENDOTHELIAL CELL INJURY IS CAUSED BY FREE RADICALS.
Anoxic insult to brain microvascular endothelial cells induced intracellular LDH leakage. This anoxic cell damage was protected by catalase and superoxide dismutase. Therefore, the endothelial cell damage is caused by free radicals from the endothelial cells themselves. NO synthetase inhibitor protected the endothelial cells from anoxic injury. It indicates NO producted in the endothelial cells may play a role as free radical source in anoxic condition.
(4) INTRACELLULAR CALCIUM PLAY AN IMPORTANT ROLE IN REVERSIBLE HYPEROSMOTIC OPENING OF BLOOD BRAIN BARRIER.
Rapid increase of intracellular calcium induced by the application of hyperosmotic mannitol was proved by confocal laser scanning microscope. The increased intracellular calcium returned to the basal level in 30 minutes. The time profile of the intracellular calcium change induced by hyperosmotic treatment coincided with the time profile of permeability change of brain capillary. It indicates the important role of intracellular calcium ion in osmotic opening of blood-brain-barrier.
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