1996 Fiscal Year Annual Research Report

脳血管関門形成の分子機構,特にアストロサイド分泌因子の解析

Research Project

Project/Area Number	08457075
Research Category	Grant-in-Aid for Scientific Research (B)
Research Institution	Sapporo Medical University
Principal Investigator	澤田典均札幌医科大学, 医学部, 助教授 (30154149)
Keywords	脳血管関門 / タイト結合 / 脳血管内皮細胞 / アストロサイト / 共焦点レーザー顕微鏡 / ZO-1 / 7H6 / 細胞電気抵抗
Research Abstract	1.目的中枢神経系の毛細血管は一層の内皮細胞とその外側を取り囲む基底膜からなり、さらにその外周をアストロサイトの足突起が覆っている。内皮細胞は、神経系と循環系との間の物質の輸送を厳密に制限する血液脳関門(blood-brain barrier:BBB)を形成し、中枢神経系の恒常性を維持するために機能している。この関門には、隣接する内皮細胞同士のタイト結合が関与していると考えられている。本研究の目的は、アストロサイトがBBBに及ぼす影響を明らかにすることにある。 2.方法と結果ブタ脳毛細血管内皮細胞の培養系を用いて、cAMPおよび星状膠細胞馴らし培地によりバリア機能の変化と、タイト結合蛋白であるZO-1,7H6抗原の発現及び細胞骨格蛋白アクチンの分布の変化と観察・検討し、さらに7H6抗原の半定量として共焦点レーザー顕微鏡による解析を行った。培地にcAMPとRO20-1724を添加したところ、添加後10時間でTERは対照群の約7倍に上昇した。cAMPとRO02-1724、および50%astrocyte conditioned medium (CM)を添加したところ、10時間後のTERは約13倍に上昇した。astrocyte CMのみを添加した群では対照群に比してTERの変化は見られなかった。一方、astrocyte CMの内皮細胞の培地に添加すると、細胞質内に7H6抗原が発現され、さらにcAMPとastrocyte CMの両者を添加することによって細胞境界域に局在が変化した。以上のことから脳毛細血管内皮細胞のタイト結合のの発達に7H6抗原の発現が深く関与していることが示唆された。

Research Products

(9 results)

All Other

All Publications (9 results)

[Publications] Satoh,H.et al: "Localization of 7H6 tight junction-associated antigen along the cell border of vascular endothelial cells correlates with paracellular barrier function against ions,large molecules and cancer cells." Exp.Cell.Res.222. 269-274 (1996)
[Publications] Tobioka,H.et al.: "Enhancced paracellular barrier function of rat mesothelial' cells partially protects against cancer cell penetration." Brt.J.Cancer. 74. 439-445 (1996)
[Publications] Kimura,M.et al.: "Comparison between the distribution of 7H6 tight junction-associated antigen and occludin during the development of chick intestine." Cell Strut.Funct.21. 91-96 (1996)
[Publications] Kato,J.et al: "Hepatic iron deprivation prevents spontaneous development of fulminant hepatitis and liver cancer in LEC rats." J.Clin.Invest.98. 923-929 (1996)
[Publications] Obata,H.et al: "Abnormal accumulation of copper in LEC rat liver induces expression of p53 and nuclear matrix-bound p21^<qwai1/cip1>." Carcinogenesis. 17. 2157-2161 (1996)
[Publications] Sawada,N.et al: "Expression of p21^<wai-1/cip-1> is significantly induced in the livers of LEC rats with chronic liver injury." Jpn.J.Cancer Res.87. 1102-1105 (1996)
[Publications] Sawada,N.et al.: "p21^<waf-1/cip-1/adi-1> is expressed at G1 phase in primary culture of hepatocytes from old rats,presumably preventing the cells from entering the S phase of the cell cycle." Biochem.Biophys.Res.Commun.228. 819-824 (1996)
[Publications] Kimura,H.et al.: "Bacterial lipopolysaccharide reduced intestinal barrier function and altered localization of 7H6 antigen in IEC-6 rat intestinal crypt cells." J.Cell.Physiol.,. (in press).
[Publications] Yamada-Sasamori,Y.et al.: "Rapid induction of 7H6 ti8ght junction-associated protein and paracellular barrier function in capillary endothelial cells of porcine brain in vitro by treatment with astrocyte conditioned medium and cAMP." Med.Electr.Microsc.,. (in press).

1996 Fiscal Year Annual Research Report

脳血管関門形成の分子機構,特にアストロサイド分泌因子の解析

Principal Investigator

澤田 典均 札幌医科大学, 医学部, 助教授 (30154149)

Research Products

[Publications] Satoh,H.et al: "Localization of 7H6 tight junction-associated antigen along the cell border of vascular endothelial cells correlates with paracellular barrier function against ions,large molecules and cancer cells." Exp.Cell.Res.222. 269-274 (1996)

[Publications] Tobioka,H.et al.: "Enhancced paracellular barrier function of rat mesothelial' cells partially protects against cancer cell penetration." Brt.J.Cancer. 74. 439-445 (1996)

[Publications] Kimura,M.et al.: "Comparison between the distribution of 7H6 tight junction-associated antigen and occludin during the development of chick intestine." Cell Strut.Funct.21. 91-96 (1996)

[Publications] Kato,J.et al: "Hepatic iron deprivation prevents spontaneous development of fulminant hepatitis and liver cancer in LEC rats." J.Clin.Invest.98. 923-929 (1996)

[Publications] Obata,H.et al: "Abnormal accumulation of copper in LEC rat liver induces expression of p53 and nuclear matrix-bound p21^<qwai1/cip1>." Carcinogenesis. 17. 2157-2161 (1996)

[Publications] Sawada,N.et al: "Expression of p21^<wai-1/cip-1> is significantly induced in the livers of LEC rats with chronic liver injury." Jpn.J.Cancer Res.87. 1102-1105 (1996)

[Publications] Sawada,N.et al.: "p21^<waf-1/cip-1/adi-1> is expressed at G1 phase in primary culture of hepatocytes from old rats,presumably preventing the cells from entering the S phase of the cell cycle." Biochem.Biophys.Res.Commun.228. 819-824 (1996)

[Publications] Kimura,H.et al.: "Bacterial lipopolysaccharide reduced intestinal barrier function and altered localization of 7H6 antigen in IEC-6 rat intestinal crypt cells." J.Cell.Physiol.,. (in press).

[Publications] Yamada-Sasamori,Y.et al.: "Rapid induction of 7H6 ti8ght junction-associated protein and paracellular barrier function in capillary endothelial cells of porcine brain in vitro by treatment with astrocyte conditioned medium and cAMP." Med.Electr.Microsc.,. (in press).

澤田典均札幌医科大学, 医学部, 助教授 (30154149)