2002 Fiscal Year Annual Research Report

メサンギウム細胞におけるAM・PAMPの作用機構の電気生理学的手法を用いた解析

Research Project

Project/Area Number	13770615
Research Institution	University of Occupational and Environmental Health, Japan
Principal Investigator	椛島成利産業医科大学, 医学部, 助手 (80279322)
Keywords	アドレノメデュリン(AM) / Ca^<2+>電流 / proadrenomedullin N-terminal 20 peptide (PAMP) / MCP-1 / MAP kinase / Patch-clamp
Research Abstract	培養メサンギウム細胞においてPatch-clamp法を応用するためにまず神経細胞においてCa^<2+>電流の基本的性質を検討した。急性単離した視索上核(SON)神経細胞を用いWhole-cell clamp modeでCa^<2+>電流を解析した。Ca^<2+>電流の単離には次の方法を用いた。TTXを灌流液に加え投与するか、または細胞外Na^+イオンを除去した灌流液にて実験することによりNa^+電流を除外し、電極内からTEAを細胞内に潅流することでK^+電流を除外した。本研究によりCa^<2+>電流は電位依存性を呈すること、さらにはcAMPの細胞外からの投与により抑制を示す事が認められた。その機序にはcAMPが最終的にadenosineに代謝されG蛋白であるGi/oを介して抑制性に作用することが判明した。この結果よりCa^<2+>電流の測定が可能であることを確認の上培養メサンギウム細胞について応用した。メサンギウム細胞において前述の条件で電位依存性Ca^<2+>電流を測定したが、電位依存性Ca^<2+>電流は極めて小さいために実験条件をCa^<2+>よりBa^<2+>に変更し電流量を増幅し解析した。しかしBa^<2+>電流も50pA程度であり、視索上核(SON)神経細胞と比較すると約1/20程度しか認められなかった。このBa^<2+>電流にアドレノメデュリン(AM)/proadrenomedullin N-terminal 20 peptide (PAMP)を投与した。しかし、電流が小さいためにAM/PAMPの作用は残念ながら判断不可能であった。さらにメサンギウム細胞の有するCa^<2+>電流を賦活化させることを念頭におき、今後の更なる研究を継続したい。またこのAMの臨床的意義に関しては同じ降圧利尿作用を有するANP, BNPと成熟型AM(mAM)の関連を血液透析患者に対して検討した。mAMの増加はANP, BNPと同様に体液量の増加を反映していることが示唆された。さらに血液透析患者においてAMの体内動態はBNPに近似することが判明した。さらに培養メサンギウム細胞の実験については糸球体高血圧が細胞増殖に関与するか否かについてケモカインであるMCP-1とMAP kinaseの関連についてRT-PCR法を用いて検討した。メサンギウム細胞を圧負荷チャンバー内で培養するとMAP kinaseを介してMCP-1のmRNAが発現することが判明した。これは糸球体高血圧がMCP-1等のケモカインを介して腎硬化症等の腎疾患に関与していることが示唆された。

Research Products

(14 results)

All Other

All Publications (14 results)

[Publications] Suda T: "Pressure induces expression of monocyte chemoattractant protein-1 through activation of MAP kinase in cultured mesangial cells"Kidney International. 60. 1705-1715 (2001)
[Publications] Iwamoto M: "Adrenomedullin inhibits pressure-induced mesangial MCP-1 expression through activation of protein kinase A"Nephrology Dialysis and Transplantation. 16・6. A15 (2001)
[Publications] Ota T: "Tubular expression of monocyte chemoattractant protein-1 (MCP-1) in rat 5/6 nephrectomy model"Nephrology Dialysis and Transplantation. 16・6. A15 (2001)
[Publications] Osajima A: "Comparison of mature adrenomedullin levels and natriuretic peptides levels as a useful marker for cardiac function in plasma of hemodialysis patients with coronary artery disease"Journal of American Society of Nephrology. 12. 405A (2001)
[Publications] Tamura M: "High glucose inhibits cell spreading, migration, and focal adhesions in peritoneal mesothelial cells"Journal of American Society of Nephrology. 12. 718A (2001)
[Publications] Watanabe Y: "Integrin-mediated cell adhesion induces monocyte chemoattractant protein-1 (MCP-1) expression in rat mesangial cells"Journal of American Society of Nephrology. 12. 720A (2001)
[Publications] Ota T: "Expression of monocyte chemoattractant protein-1 (MCP-1) in tubulointerstitial lesions of subtotal nephrectomy rats"Journal of American Society of Nephrology. 12. 822A (2001)
[Publications] Suda T: "The mature form of adrenomedullin correlates with brain natriuretic peptide in plasma of chronic hemodialysis patients"Clinical Nephrology. 57. 444-451 (2002)
[Publications] Osajima A: "Comparison of plasma levels of mature adrenomedullin and natriuretic peptide as markers of cardiac function in hemodialysis patients with coronary artery disease"Nephron. 92. 832-839 (2002)
[Publications] Tamura M: "High glucose inhibits focal adhesion kinase (FAK)-mediated wound healing of peritoneal mesothelial cells"Nephrology Dialysis and Transplantation. 17. S300 (2002)
[Publications] Watanabe Y: "Integrin-mediated cell adhesion induces monocyte chemoattractant protein-1 (MCP-1) mRNA expression in cultured rat mesangial cells"Nephrology Dialysis and Transplantation. 17. S71 (2002)
[Publications] Osajima A: "Mature adrenomedullin levels are less useful than natriuretic peptides levels as a marker of cardiac function in hemodialysis patients with coronary artery disease"Nephrology Dialysis and Transplantation. 17. S262 (2002)
[Publications] Tamura M: "High glucose concentrations inhibits focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells"Journal of American Society of Nephrology. 13. 525A (2002)
[Publications] Tamura M: "High glucose levels inhibit focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells. Kidney Int.63:722-731"Kidney International. 63. 722-731 (2003)

2002 Fiscal Year Annual Research Report

メサンギウム細胞におけるAM・PAMPの作用機構の電気生理学的手法を用いた解析

Principal Investigator

椛島 成利 産業医科大学, 医学部, 助手 (80279322)

Research Products

[Publications] Suda T: "Pressure induces expression of monocyte chemoattractant protein-1 through activation of MAP kinase in cultured mesangial cells"Kidney International. 60. 1705-1715 (2001)

[Publications] Iwamoto M: "Adrenomedullin inhibits pressure-induced mesangial MCP-1 expression through activation of protein kinase A"Nephrology Dialysis and Transplantation. 16・6. A15 (2001)

[Publications] Ota T: "Tubular expression of monocyte chemoattractant protein-1 (MCP-1) in rat 5/6 nephrectomy model"Nephrology Dialysis and Transplantation. 16・6. A15 (2001)

[Publications] Osajima A: "Comparison of mature adrenomedullin levels and natriuretic peptides levels as a useful marker for cardiac function in plasma of hemodialysis patients with coronary artery disease"Journal of American Society of Nephrology. 12. 405A (2001)

[Publications] Tamura M: "High glucose inhibits cell spreading, migration, and focal adhesions in peritoneal mesothelial cells"Journal of American Society of Nephrology. 12. 718A (2001)

[Publications] Watanabe Y: "Integrin-mediated cell adhesion induces monocyte chemoattractant protein-1 (MCP-1) expression in rat mesangial cells"Journal of American Society of Nephrology. 12. 720A (2001)

[Publications] Ota T: "Expression of monocyte chemoattractant protein-1 (MCP-1) in tubulointerstitial lesions of subtotal nephrectomy rats"Journal of American Society of Nephrology. 12. 822A (2001)

[Publications] Suda T: "The mature form of adrenomedullin correlates with brain natriuretic peptide in plasma of chronic hemodialysis patients"Clinical Nephrology. 57. 444-451 (2002)

[Publications] Osajima A: "Comparison of plasma levels of mature adrenomedullin and natriuretic peptide as markers of cardiac function in hemodialysis patients with coronary artery disease"Nephron. 92. 832-839 (2002)

[Publications] Tamura M: "High glucose inhibits focal adhesion kinase (FAK)-mediated wound healing of peritoneal mesothelial cells"Nephrology Dialysis and Transplantation. 17. S300 (2002)

[Publications] Watanabe Y: "Integrin-mediated cell adhesion induces monocyte chemoattractant protein-1 (MCP-1) mRNA expression in cultured rat mesangial cells"Nephrology Dialysis and Transplantation. 17. S71 (2002)

[Publications] Osajima A: "Mature adrenomedullin levels are less useful than natriuretic peptides levels as a marker of cardiac function in hemodialysis patients with coronary artery disease"Nephrology Dialysis and Transplantation. 17. S262 (2002)

[Publications] Tamura M: "High glucose concentrations inhibits focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells"Journal of American Society of Nephrology. 13. 525A (2002)

[Publications] Tamura M: "High glucose levels inhibit focal adhesion kinase-mediated wound healing of rat peritoneal mesothelial cells. Kidney Int.63:722-731"Kidney International. 63. 722-731 (2003)

椛島成利産業医科大学, 医学部, 助手 (80279322)