1990 Fiscal Year Annual Research Report

コレチストキニン放出ペプチドの精製及びその生理的役割

Research Project

Project/Area Number	02670332
Research Institution	Tokyo Metropolitan Institute of Gerontology
Principal Investigator	宮坂京子東京都老人総合研究所, 臨床生理部門, 研究員 (90166140)
Co-Investigator(Kenkyū-buntansha)	船越顕博国立病院九州がんセンター, 内科, 医長
Keywords	コレチストキニン / 膵外分泌 / 放出ペプチド / トリプシン
Research Abstract	ラット小腸を幽門から20cmのところで切断し、内腔を洗浄後、縦方向に切開し、腸粘膜をかきとって集め、約100gを一回分とした。これまでに5回にわたり小腸粘膜の採取とそこからのCCKーRPの抽出と精製を行った。採取した粘膜1gあたり1m1の1M酢酸を加え、10分間煮沸後、ホモジナイズし15,000rpmで30分遠心した。上澄みをSephade×G50カラムを用いゲル濾過し、覚醒ラットを用いたモデルで生物活性(膵外分泌刺激作用とCCK放出能)を検討し、活性を有する分画をHPLCによりさらに精製するということを繰り返し行った。HPLCは、Cosmosil Cー18カラムを用い、10ー60%アセトニトリル平衡濃度勾配により分離した。この条件下では生物活性を持つ分画が単一のpeakとなったのでアミノ酸分析を試みたが、自動分析ができなかった。ここまでの時点で、この物質は熱に対し安定で、分子量が1000ー10000の間にあり、トリプシンとのincubationにより活性を失い、それ自身はトリプシンインヒビタ-活性を持たないと言うことがわかり、初めに想定したものとその性格はほぼ一致した。このpeakの生物活性を膵液由来のCCKーRPと考えられるmonitor peptideと比較したところ、殆ど同じ力価を呈していた。その後、さらにHPLCのカラムや条件を替えてこの分画の精製を繰り返したところ、おおきなpeakと一見コンタミ様の5つ計6つの分画に分離された、思いがけず、このうちの最もおおきなpeakには生物活性が認められず、コンタミかとおもわれた小さなpeakに活性が認められた。現在このpeakをさらに精製している最中である。

Research Products

(25 results)

All Other

All Publications (25 results)

[Publications] 中村理恵子: "Luminal bile regulates cholecystokinin release in conscious rats." Dig.Dis.Sci.35. 55-60 (1990)
[Publications] 宮坂京子: "Rat pancreastatin inhibits both pancreatic exocrine and endocrine secretions in rats." Regul.Pept.28. 131-240 (1990)
[Publications] 宮坂京子: "The importance of Cーterminal amide structure of rat pancreastatin to inhibit pancreatic exocrine secretion." FEBS Lett.263. 279-280 (1990)
[Publications] 鹿戸福子: "Necessity of hyperglycemia for effects of endogenous cholecystokinin on insulin and pancreatic exocrine secretion in conscious rats." Jap.J.Physiol.40. 383-391 (1990)
[Publications] 船越顕博: "Plasma pancreastatin responses after intraduodenal infusion of liquid meal in patients with chronic pancreatitis." Dig.Dis.Sci.35. 721-725 (1990)
[Publications] Tamamura,H: "Isolation and characterization of a tumorーderived human chromogranin Aーrelated protein and its invitro conversion to human pancreastatinー48." Eurp.J.Biochem.191. 33-39 (1990)
[Publications] 立石かよ子: "Pancreastatinーlike immunoreactivity in urine." J.Clin.Endocrinol.Metab.71. 842-845 (1990)
[Publications] 宮坂京子: "Inhibitory effect of pancreastatin on pancreatic exocrine secretions:Pancreastatin inhibits central vagal nerve stimulation." Gastroenterology. 99. 1751-1756 (1990)
[Publications] 篠崎博嗣: "Bioactivity of synthetic human cholecystokinin(CCK)ー33 in vitro and in vivo." Gastroenterologia Japonica. 26. 51-55 (1990)
[Publications] 宮坂京子: "Bile acids in human plasma interfere with cholecystokinin bioassay using dispersed pancreatic acini." Dig.Dis.Sci.
[Publications] 木村理: "Immunohistochemical study of the distribution of pancreastatin in endocrine tumores of the pancreas and normal pancreatic tissue:Analysis of autopsy cases." Pancreas.
[Publications] 自見厚郎: "Tubular complex in the pancreatic tissue after pancreatic duct ligation in rats." J.Clin.Electron Microscopy.
[Publications] 宮坂京子: "Absence of luminal bile increases duodenal content of cholecyosokinin in rats." Proc.Soc.Exp.Biol.Med.
[Publications] 宮坂京子: "Inhibitory effect of taurocholate on CCK release and pancreatic secretion produced by two CCKーreleasing peptide in conscious rats." Am.J.Physiol.revised version submitted.
[Publications] 宮坂京子: "Stimulatory and inhibitory effects of bile satls on rat pancreatic secretion." Gastroenterology.
[Publications] 宮坂京子: "Changes in plasma and duodenal cholecystokinin concentrations after pancreatic duct occlusion in rats." Dig.Dis.Sci.
[Publications] 宮坂京子: "Protective effect of human pancreatic secretory trypsin inhibitor on cerulein induced acute pancreatitis." Pancreas,.
[Publications] 宮坂京子: "Extraction of CCKーreleasing peptide from rat intestine and its bioactivity."
[Publications] 自見厚郎: "高脂肪食飼育ラットのセルレイン誘発膵炎の防御効果。" 膵臓. 5. 79-88 (1990)
[Publications] 船越顕博: "ラットのセルレイン膵炎に対するcamostatの予防効果。" 現代医療. 22. 538-545 (1990)
[Publications] 宮坂京子: "各種胆汁酸の膵外分泌の対する作用と腸管内胆汁酸(タウロコ-ル酸)の役割。" 消化と吸収. 13. 46-49 (1990)
[Publications] 船越顕博: "新しいペプタイド"パンクレアスタチン"のMeal負荷時の膵内外分泌に対する効果。" 第40回ペプシン研究会議事録。. 128-136 (1990)
[Publications] 船越顕博: "腫瘍由来のヒトpancreastatin関連蛋白の分離精製と性質。" Peptide Hormones in Pancreas.10. 308-312 (1990)
[Publications] 船越顕博: "Effect of pancreastatin on pancreatic endocrine function in the rat(in vivo and in virto study).Recent trends of diabetes mellitus in east asia.(分担)" Elsevier^ー Sciences Publishers,Amsterdam,(1990)
[Publications] 宮坂京子: "消化管ホルモンIX.(分担)" 医学図書出版、, 582 (1990)

1990 Fiscal Year Annual Research Report

コレチストキニン放出ペプチドの精製及びその生理的役割

Principal Investigator

宮坂 京子 東京都老人総合研究所, 臨床生理部門, 研究員 (90166140)

Research Products

[Publications] 中村 理恵子: "Luminal bile regulates cholecystokinin release in conscious rats." Dig.Dis.Sci.35. 55-60 (1990)

[Publications] 宮坂 京子: "Rat pancreastatin inhibits both pancreatic exocrine and endocrine secretions in rats." Regul.Pept.28. 131-240 (1990)

[Publications] 宮坂 京子: "The importance of Cーterminal amide structure of rat pancreastatin to inhibit pancreatic exocrine secretion." FEBS Lett.263. 279-280 (1990)

[Publications] 鹿戸 福子: "Necessity of hyperglycemia for effects of endogenous cholecystokinin on insulin and pancreatic exocrine secretion in conscious rats." Jap.J.Physiol.40. 383-391 (1990)

[Publications] 船越 顕博: "Plasma pancreastatin responses after intraduodenal infusion of liquid meal in patients with chronic pancreatitis." Dig.Dis.Sci.35. 721-725 (1990)

[Publications] Tamamura,H: "Isolation and characterization of a tumorーderived human chromogranin Aーrelated protein and its invitro conversion to human pancreastatinー48." Eurp.J.Biochem.191. 33-39 (1990)

[Publications] 立石 かよ子: "Pancreastatinーlike immunoreactivity in urine." J.Clin.Endocrinol.Metab.71. 842-845 (1990)

[Publications] 宮坂 京子: "Inhibitory effect of pancreastatin on pancreatic exocrine secretions:Pancreastatin inhibits central vagal nerve stimulation." Gastroenterology. 99. 1751-1756 (1990)

[Publications] 篠崎 博嗣: "Bioactivity of synthetic human cholecystokinin(CCK)ー33 in vitro and in vivo." Gastroenterologia Japonica. 26. 51-55 (1990)

[Publications] 宮坂 京子: "Bile acids in human plasma interfere with cholecystokinin bioassay using dispersed pancreatic acini." Dig.Dis.Sci.

[Publications] 木村 理: "Immunohistochemical study of the distribution of pancreastatin in endocrine tumores of the pancreas and normal pancreatic tissue:Analysis of autopsy cases." Pancreas.

[Publications] 自見 厚郎: "Tubular complex in the pancreatic tissue after pancreatic duct ligation in rats." J.Clin.Electron Microscopy.

[Publications] 宮坂 京子: "Absence of luminal bile increases duodenal content of cholecyosokinin in rats." Proc.Soc.Exp.Biol.Med.

[Publications] 宮坂 京子: "Inhibitory effect of taurocholate on CCK release and pancreatic secretion produced by two CCKーreleasing peptide in conscious rats." Am.J.Physiol.revised version submitted.

[Publications] 宮坂 京子: "Stimulatory and inhibitory effects of bile satls on rat pancreatic secretion." Gastroenterology.

[Publications] 宮坂 京子: "Changes in plasma and duodenal cholecystokinin concentrations after pancreatic duct occlusion in rats." Dig.Dis.Sci.

[Publications] 宮坂 京子: "Protective effect of human pancreatic secretory trypsin inhibitor on cerulein induced acute pancreatitis." Pancreas,.

[Publications] 宮坂 京子: "Extraction of CCKーreleasing peptide from rat intestine and its bioactivity."

[Publications] 自見 厚郎: "高脂肪食飼育ラットのセルレイン誘発膵炎の防御効果。" 膵臓. 5. 79-88 (1990)

[Publications] 船越 顕博: "ラットのセルレイン膵炎に対するcamostatの予防効果。" 現代医療. 22. 538-545 (1990)

[Publications] 宮坂 京子: "各種胆汁酸の膵外分泌の対する作用と腸管内胆汁酸(タウロコ-ル酸)の役割。" 消化と吸収. 13. 46-49 (1990)

[Publications] 船越 顕博: "新しいペプタイド"パンクレアスタチン"のMeal負荷時の膵内外分泌に対する効果。" 第40回ペプシン研究会議事録。. 128-136 (1990)

[Publications] 船越 顕博: "腫瘍由来のヒトpancreastatin関連蛋白の分離精製と性質。" Peptide Hormones in Pancreas.10. 308-312 (1990)

[Publications] 船越 顕博: "Effect of pancreastatin on pancreatic endocrine function in the rat(in vivo and in virto study).Recent trends of diabetes mellitus in east asia.(分担)" Elsevier^ー Sciences Publishers,Amsterdam,(1990)

[Publications] 宮坂 京子: "消化管 ホルモンIX.(分担)" 医学図書出版、, 582 (1990)

宮坂京子東京都老人総合研究所, 臨床生理部門, 研究員 (90166140)

[Publications] 中村理恵子: "Luminal bile regulates cholecystokinin release in conscious rats." Dig.Dis.Sci.35. 55-60 (1990)

[Publications] 宮坂京子: "Rat pancreastatin inhibits both pancreatic exocrine and endocrine secretions in rats." Regul.Pept.28. 131-240 (1990)

[Publications] 宮坂京子: "The importance of Cーterminal amide structure of rat pancreastatin to inhibit pancreatic exocrine secretion." FEBS Lett.263. 279-280 (1990)

[Publications] 鹿戸福子: "Necessity of hyperglycemia for effects of endogenous cholecystokinin on insulin and pancreatic exocrine secretion in conscious rats." Jap.J.Physiol.40. 383-391 (1990)

[Publications] 船越顕博: "Plasma pancreastatin responses after intraduodenal infusion of liquid meal in patients with chronic pancreatitis." Dig.Dis.Sci.35. 721-725 (1990)

[Publications] 立石かよ子: "Pancreastatinーlike immunoreactivity in urine." J.Clin.Endocrinol.Metab.71. 842-845 (1990)

[Publications] 宮坂京子: "Inhibitory effect of pancreastatin on pancreatic exocrine secretions:Pancreastatin inhibits central vagal nerve stimulation." Gastroenterology. 99. 1751-1756 (1990)

[Publications] 篠崎博嗣: "Bioactivity of synthetic human cholecystokinin(CCK)ー33 in vitro and in vivo." Gastroenterologia Japonica. 26. 51-55 (1990)

[Publications] 宮坂京子: "Bile acids in human plasma interfere with cholecystokinin bioassay using dispersed pancreatic acini." Dig.Dis.Sci.

[Publications] 木村理: "Immunohistochemical study of the distribution of pancreastatin in endocrine tumores of the pancreas and normal pancreatic tissue:Analysis of autopsy cases." Pancreas.

[Publications] 自見厚郎: "Tubular complex in the pancreatic tissue after pancreatic duct ligation in rats." J.Clin.Electron Microscopy.

[Publications] 宮坂京子: "Absence of luminal bile increases duodenal content of cholecyosokinin in rats." Proc.Soc.Exp.Biol.Med.

[Publications] 宮坂京子: "Inhibitory effect of taurocholate on CCK release and pancreatic secretion produced by two CCKーreleasing peptide in conscious rats." Am.J.Physiol.revised version submitted.

[Publications] 宮坂京子: "Stimulatory and inhibitory effects of bile satls on rat pancreatic secretion." Gastroenterology.

[Publications] 宮坂京子: "Changes in plasma and duodenal cholecystokinin concentrations after pancreatic duct occlusion in rats." Dig.Dis.Sci.

[Publications] 宮坂京子: "Protective effect of human pancreatic secretory trypsin inhibitor on cerulein induced acute pancreatitis." Pancreas,.

[Publications] 宮坂京子: "Extraction of CCKーreleasing peptide from rat intestine and its bioactivity."

[Publications] 自見厚郎: "高脂肪食飼育ラットのセルレイン誘発膵炎の防御効果。" 膵臓. 5. 79-88 (1990)

[Publications] 船越顕博: "ラットのセルレイン膵炎に対するcamostatの予防効果。" 現代医療. 22. 538-545 (1990)

[Publications] 宮坂京子: "各種胆汁酸の膵外分泌の対する作用と腸管内胆汁酸(タウロコ-ル酸)の役割。" 消化と吸収. 13. 46-49 (1990)

[Publications] 船越顕博: "新しいペプタイド"パンクレアスタチン"のMeal負荷時の膵内外分泌に対する効果。" 第40回ペプシン研究会議事録。. 128-136 (1990)

[Publications] 船越顕博: "腫瘍由来のヒトpancreastatin関連蛋白の分離精製と性質。" Peptide Hormones in Pancreas.10. 308-312 (1990)

[Publications] 船越顕博: "Effect of pancreastatin on pancreatic endocrine function in the rat(in vivo and in virto study).Recent trends of diabetes mellitus in east asia.(分担)" Elsevier^ー Sciences Publishers,Amsterdam,(1990)

[Publications] 宮坂京子: "消化管ホルモンIX.(分担)" 医学図書出版、, 582 (1990)