1995 Fiscal Year Annual Research Report

血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステムの開発

Research Project

Project/Area Number	06557045
Research Institution	Kyushu University
Principal Investigator	金出英夫九州大学, 医学部, 教授 (80038851)
Co-Investigator(Kenkyū-buntansha)	西村淳二九州大学, 医学部, 講師 (90237727) 小林誠九州大学, 医学部, 助教授 (80225515)
Keywords	血管内皮 / 血管平滑筋 / 生体計測 / 細胞質カルシウム / フラ-2
Research Abstract	1.昨年作製した「血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステム・第1号機」について、本年度は次の改良を行った。 (1)回析格子で得られる2つの励起光(340、380nm)の切替照射法として、ステッピングモーターによるチョッパーミラーの回転方式を採用した。ことによって、比較的高速(10Hz)の切替が可能となった。 (2)チョッパー切替装置の制御は画像処理装置のチョッパードライブを用いた。 (3)画像取り込みはSITカメラを用いることにした。画像処理装置のフレームおよび拡張メモリーを用いて、640×400ドットの8ビット(256階調)画像フレームを得ることができた。 (4)細胞内の[Ca]i濃度分布像について、256階調の[Ca]i濃度レベルを疑似カラーで表示可能となった。この画像は、5Hzで得ることができるようになった。 2.本システムを利用し、次のような研究成果を挙げた。 (1)in situ豚大動脈弁の内皮細胞に光学的方法を応用し、[Ca]i(フラ-2蛍光)と一酸化窒素-NO(diaminonaphthalene蛍光)の同時測定を試みた。内皮由来弛緩因子を産生するアゴニスト(ATP,Brady kinin,Thrombin)は、内皮細胞に用量依存性の[Ca]i上昇とNO産生を引き起こした。[Ca]i変化-NO産生量の関係は、アゴニストの種類によって異なることが明らかとなった。 (2)冠動脈および腎動脈におけるアドレノメジュリン(AM)の産生と機能について検討した。K脱分極やエンドセリン1で前収縮させた冠動脈や腎動脈中膜標本の[Ca]iと張力を、AMは減少させた。これらの標本にReverse transcription polymerase chain reaction法を応用し、AM mRNAの発現を証明した。冠動脈および腎動脈において、AMはオートクリン/パラクリン型の血管緊張調節機能を有することが明らかとなった。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] M.Ushio-Fukai: "Cytosolic calcium concentration-force relation during contractions in the rabbit femoral artery" Br J Pharmacol. 114. 329-338 (1995)
[Publications] Y.Hirakawa: "Dual regulation of L-type Ca^<2+> channels by serotonin 2 recepror stimulation in vascular smooth muscle cells" Am J Physiol. 268. H544-H549 (1995)
[Publications] Y.Miyagi: "Resting load regulates cytosolic calcium-force relationship of the contraction of bovine cerebrovascular smooth muscle" J Physiol. 484. 123-137 (1995)
[Publications] Y.Miyagi: "Resting load regulates vascular sensitivity by a cytosolic Ca^<2+> -insensitive mechaism" Am J Physiol. 268. C1332-C1341 (1995)
[Publications] K.Toyofuku: "Effects of U46619 on intracellular Ca^<++> concentration and tension in human umbilical artery" Am J Obstet Gynecol. 172. 1414-1421 (1995)
[Publications] G.Hiroishi: "High-D-glucose stimulates the cell Cycle from the G1 to S and M phases,but has no competent effect on the G0 phase,in vascular" Biochem biophys Res Commun. 211. 619-629 (1995)
[Publications] Y.Kureishi: "Adrenomedullin decreases both cytosolic Ca^<2+> concentration and Ca^<2+> sensitivity in pig coronary arterial smooth muscle." Biochem Biophys Res Commun. 212. 572-579 (1995)
[Publications] J.Nishimura: "Evidence for the presence of endothelin ET_A receptors in endothelial cells in situ of aortic side of porcine aortic valve" Br J Pharmacol. 115. 1369-1376 (1995)
[Publications] M.Kuroiwa: "The mechanism of endothelium-dependent relaxation induced by substance P in the porcine coronary artery" Br J Pharmacol. 116. 2040-2047 (1995)
[Publications] H.Seguchi: "Autocrine regulation of the renal arterial tone by adrenomedullin" Biochem Biophys Res Commun. 215. 619-625 (1995)
[Publications] H.Yoshimura: "The effects of midazolam on intracellular Ca^<2+> and tension in airway smooth muscle" Anesthesiology. 83. 1009-1020 (1995)
[Publications] H.Jahan: "Both endothelin-1 and angiotensin-II act as progression growth factors but not competence growth factors in vascular smooth muscle cells in primary culture" Eur J Pharmacol. 295. 261-269 (1996)
[Publications] Y.Miyagi: "Nucleotide receptor links to cytosolic calcium transient and release of vasorelaxint factor in bovine endothelial cells in situ" J Pyhsiol. (in press).
[Publications] Y.Higuchi: "Cyclopiazonic acid induces a sustained increase in the intracellular Ca^<2+> concentration of endothelial cells on the porcine aortic valve and a relaxation of the porcine coronary artery" Am J Physiol. (in press).
[Publications] S.Abe: "Some effects of nipradilol,a β-blocker possessing anitroxy group,on smooth muscle of the pig coronary artery" Br J Pharmacol. (in press).

1995 Fiscal Year Annual Research Report

血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステムの開発

Principal Investigator

金出 英夫 九州大学, 医学部, 教授 (80038851)

Research Products

[Publications] M.Ushio-Fukai: "Cytosolic calcium concentration-force relation during contractions in the rabbit femoral artery" Br J Pharmacol. 114. 329-338 (1995)

[Publications] Y.Hirakawa: "Dual regulation of L-type Ca^<2+> channels by serotonin 2 recepror stimulation in vascular smooth muscle cells" Am J Physiol. 268. H544-H549 (1995)

[Publications] Y.Miyagi: "Resting load regulates cytosolic calcium-force relationship of the contraction of bovine cerebrovascular smooth muscle" J Physiol. 484. 123-137 (1995)

[Publications] Y.Miyagi: "Resting load regulates vascular sensitivity by a cytosolic Ca^<2+> -insensitive mechaism" Am J Physiol. 268. C1332-C1341 (1995)

[Publications] K.Toyofuku: "Effects of U46619 on intracellular Ca^<++> concentration and tension in human umbilical artery" Am J Obstet Gynecol. 172. 1414-1421 (1995)

[Publications] G.Hiroishi: "High-D-glucose stimulates the cell Cycle from the G1 to S and M phases,but has no competent effect on the G0 phase,in vascular" Biochem biophys Res Commun. 211. 619-629 (1995)

[Publications] Y.Kureishi: "Adrenomedullin decreases both cytosolic Ca^<2+> concentration and Ca^<2+> sensitivity in pig coronary arterial smooth muscle." Biochem Biophys Res Commun. 212. 572-579 (1995)

[Publications] J.Nishimura: "Evidence for the presence of endothelin ET_A receptors in endothelial cells in situ of aortic side of porcine aortic valve" Br J Pharmacol. 115. 1369-1376 (1995)

[Publications] M.Kuroiwa: "The mechanism of endothelium-dependent relaxation induced by substance P in the porcine coronary artery" Br J Pharmacol. 116. 2040-2047 (1995)

[Publications] H.Seguchi: "Autocrine regulation of the renal arterial tone by adrenomedullin" Biochem Biophys Res Commun. 215. 619-625 (1995)

[Publications] H.Yoshimura: "The effects of midazolam on intracellular Ca^<2+> and tension in airway smooth muscle" Anesthesiology. 83. 1009-1020 (1995)

[Publications] H.Jahan: "Both endothelin-1 and angiotensin-II act as progression growth factors but not competence growth factors in vascular smooth muscle cells in primary culture" Eur J Pharmacol. 295. 261-269 (1996)

[Publications] Y.Miyagi: "Nucleotide receptor links to cytosolic calcium transient and release of vasorelaxint factor in bovine endothelial cells in situ" J Pyhsiol. (in press).

[Publications] Y.Higuchi: "Cyclopiazonic acid induces a sustained increase in the intracellular Ca^<2+> concentration of endothelial cells on the porcine aortic valve and a relaxation of the porcine coronary artery" Am J Physiol. (in press).

[Publications] S.Abe: "Some effects of nipradilol,a β-blocker possessing anitroxy group,on smooth muscle of the pig coronary artery" Br J Pharmacol. (in press).

金出英夫九州大学, 医学部, 教授 (80038851)