OCTNで取り込まれる新規細菌活性物質の腸上皮細胞内動態とMDR1による排出障害

2009 Fiscal Year Annual Research Report

• Project/Area Number: 20590734
• Research Institution: Asahikawa Medical College
• Principal Investigator: 藤谷 幹浩 Asahikawa Medical College, 医学部, 准教授 (80322915)
• Co-Investigator(Kenkyū-buntansha): 大竹 孝明 旭川医科大学, 医学部, 講師 (10359490)
• Keywords: MDR-1 / OCTN / Bacillus subtilis / Competence and Sporulation factor / 炎症性腸疾患

Research Abstract

1.腸内細菌産生ペプチドの細胞内動態を明らかにする.
バシラス菌由来の腸管保護物質であるcompetence and sporulation factor(CSF)を化学的に合成しFITC標識をした後,大腸癌細胞株Caco2/bbe cellsに添加した結果,15分後に細胞質内に取りこまれることを明らかにした.その後,30-60分後にその一部は核内に取り込まれていくことも明らかにした.したがって,バシラス菌由来の活性物質であるCSFはヒト腸管上皮細胞に取り込まれ,さらに核内へと輸送されることで生理作用を発揮することが示唆された.
2.MDR-1による細菌産生ペプチドの排出能を明らかにする.
Caco2/bbe cellsにMDR-1 siRNAを導入し,MDR-1低発現細胞株を作製した.さらに,preliminaryな検討として,本細胞株に対しFITC標識CSFを添加して細胞内蓄積量を検討した結果,control細胞と比較して細胞内におけるFITCレベルの増加を認めた.すなわち,MDR-1はCSFの細胞外排泄にかかわっている可能性が示唆された.
3.MDR1の機能異常に起因する細菌産生ペプチドの蓄積と腸炎発症との関連性を明らかにする.
変異MDR-1遺伝子を作成した.現在,遺伝子導入における条件設定を行っており,次年度はこのin vivoモデルを用いて,IBD関連遺伝子異常と腸炎発症のとの関係を明らかにしていく.

Research Products

• [Journal Article] Precursor Processing of Human Defensin-5 Is Essential to the Multiple Functions in vitro and in vivo. (2010)
  • Author(s): Ishikawa C, Tanabe H, Maemoto A, Ito T, Watari J, Kono T, Fujiya M, Ashida T, Ayabe T, Kohgo Y.
  • Journal Title: J Innate Immunity 2 Pages: 66-76
  • Peer Reviewed
• [Journal Article] Small RNAsと消化管炎症性疾患の病態 (2010)
  • Author(s): 奈田利恵、藤谷幹浩、高後裕
  • Journal Title: 分子消化器病 3 Pages: 23-28
• [Journal Article] Effects of Helicobacter pylori infection on genetic instability, the aberrant CpG island methylation status and the cellular phenotype in Barrett's esophagus in a Japanese population (2009)
  • Author(s): Moriichi K, Watari J, Das KM, Tanabe H, Fujiya M, Ashida T, Kohgo Y.
  • Journal Title: Int J Cancer 124 Pages: 1263-9
  • Peer Reviewed
• [Journal Article] Immunoprecipitation of nucleosomal DNA is a novel procedure to improve the sensitivity of serum screening for the p16 hypermethylation associated with colon cancer.
  • Author(s): Sakamoto J, Fujiya M, Okamoto K, Nata T, Inaba Y, Moriichi K, Tanabe H, Mizukami Y, Watari J, Ashida T, Kohgo Y.
  • Journal Title: Cancer Epidemiology (in press)
  • Peer Reviewed
• [Journal Article] The expression of the anti-microbial peptide α-defensin/cryptdins inintestinal crypts decreases at the initial phase of intestinal inflammation in a model of inflammatory bowel disease, IL-10 deficient mice
  • Author(s): Inaba Y, Ashida T, Ito T, Ishikawa C, Tanabe H, Maemoto A, Watari J, Ayabe T, Mizukami Y, Fujiya M, Kohgo Y.
  • Journal Title: Inflammatory Bowel Diseases (in press)
  • Peer Reviewed
• [Journal Article] Obscure gastrointestinal bleeding from vascular lesions formed by venous and lymphatic congestion due to post-operative adhesion and subsequent mesenteric torsion 50 years after appendectomy.
  • Author(s): Kashima S, Nata T, Fujiya M, Moriichi K, Nomura Y, Ueno N, Itabashi K, Ishikawa C, Inaba Y, Ito T, Okamoto K, Mizukami Y, Ebisawa Y, Chisato N, Kono T, Tokusashi Y, Miyokawa N, Yamada M, Kohgo Y.
  • Journal Title: Gut (in press)
  • Peer Reviewed
• [Journal Article] 麦芽乳酸菌(L.brevis 8803)死菌の腸管組織に対する生理活性の解明
  • Author(s): 上野伸展, 瀬川修一, 藤谷幹浩, 杉山隆治, 奈田利恵, 野村好紀, 嘉島伸, 石川千里, 稲場勇平, 伊藤貴博, 盛一健太郎, 岡本耕太郎, 高後裕
  • Journal Title: 消化器と免疫 (in press)
  • Peer Reviewed
• [Presentation] Heat killed Lactobaccilus brevis SBC8803 improves intestinal injury in a murine model of colitis via the enhancement of the intestinal barrier function and the down-regulation of pro-inflammatory cytokines. (2010)
  • Author(s): Fujiya M, Ueno N, Segawa S, Nata T, Moriichi K, Tanabe H, Mizukami Y, Kobayashi N, Ito K, Kohgo Y
  • Organizer: The 4th Korea-Japan Inflammatory Bowel Disease Symposium
  • Place of Presentation: Tokyo
  • Year and Date: 2010-01-23
• [Presentation] 枯草菌(bacillus subtilis)由来のCompetence and sporulation factor(CSF)は免疫調節作用や細胞保護作用を介して腸管炎症における粘膜上皮細胞障害を改善する (2009)
  • Author(s): 岡本耕太郎、藤谷幹浩、高後裕
  • Organizer: 第47回小腸研究会
  • Place of Presentation: 福岡
  • Year and Date: 2009-11-14
• [Presentation] Heat killed Lactobacillus brevis SB8803 contributes to intestinal homeostasis via induction of heat shock proteins and activation of p38 mitogen-activated protein kinase. (2009)
  • Author(s): Ueno N, Fujiya M, Segawa S, Nata T, Inaba Y, Moriichi K, Tanabe H, Kohgo Y.
  • Organizer: Cell stress society international, 4th international conference
  • Place of Presentation: Sapporo
  • Year and Date: 2009-10-06
• [Presentation] 新規麦芽乳酸菌の腸管に対する生理活性の解明 (2009)
  • Author(s): 上野伸展, 瀬川修一, 藤谷幹浩, 杉山隆治, 奈田利恵, 野村好紀, 嘉島伸, 石川千里, 稲場勇平, 伊藤貴博, 盛一健太郎, 岡本耕太郎, 高後裕
  • Organizer: 第46回日本消化器免疫学会総会
  • Place of Presentation: 松山
  • Year and Date: 2009-07-24
• [Presentation] Infliximab may prevent post-operative recurrences in Crohn's disease. (2009)
  • Author(s): K.Okamoto, T.Ashida, R.Sugiyama, T.Nata, Y.Nomura, N.Ueno, C.Ishikawa, Y.Inaba, T.Ito, K.Moriihi, H.Tanabe, M.Fujiya, Y.Kohgo
  • Organizer: DDW 2009(AGA)
  • Place of Presentation: Chicago
  • Year and Date: 2009-06-01
• [Presentation] Bacillus subtilis-produced pentapeptide csf contributes to intestinal homeostasis via octn2, a host cell membrane transporter. (2009)
  • Author(s): Fujiya M, Inaba Y, Nata T, Moriichi K, Okamoto K, Tanabe H, Mizukami Y, Chang EB, Kohgo Y
  • Organizer: 13th Taishotoyama International Symposium on Gastroenterology
  • Place of Presentation: Shimoda
  • Year and Date: 2009-04-18
• [Patent(Industrial Property Rights)] 腸管保護剤 (2010)
  • Inventor(s): 藤谷幹浩, 高後裕, 上野伸展, 瀬川修一, 小林直之
  • Industrial Property Rights Holder: 国立大学法人旭川医科大学 サッポロビール株式会社
  • Industrial Property Number: 特願2010-089469
  • Filing Date: 2010-04-08