Intravital imaging of microcirculation on murine sepsis model using two-photon laser scanning microscopy

• Project/Area Number: 25462053
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Mie University
• Principal Investigator: KOBAYASHI MINAKO 三重大学, 医学(系)研究科(研究院), 講師 (30359765)
• Co-Investigator(Kenkyū-buntansha): KOIKE Yuki 三重大学, 医学部附属病院, 助教 (10555551) ; TOIYAMA Yuji 三重大学, 医学系研究科, 助手 (00422824) ; TANAKA Koji 三重大学, 医学部付属病院, 講師 (10345986) ; MOHRI Yasuhiko 三重大学, 医学系研究科, 准教授 (70345974) ; MIZIGUCHI Akira 三重大学, 医学系研究科, 教授 (90181916) ; KUSUNOKI Masato 三重大学, 医学系研究科, 教授 (50192026)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥260,000 (Direct Cost: ¥200,000、Indirect Cost: ¥60,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 二光子励起顕微鏡 / 敗血症 / リアルタイムイメージング / 好中球 / 末梢循環障害 / 生体内イメージンング / 消化管微小循環

Outline of Final Research Achievements

We developed a method of intravital imaging of interaction between leukocytes, platelets and vascular endothelium in tissue microcirculation of a lipopolysaccharide -induced sepsis model using a two photon laser scanning microscopy.
We imaged the rolling or adhesion of leukocytes to vascular endothelium, leukocyte-platelet aggregates, and platelet aggregation in postcapillary venules of the cecum and hepatic sinusoids in vivo real-time. We also imaged neutrophil extracellular traps (NETs) which are activated by these interactions. NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils (fragmented or cotton-like structures) and anchored NETs that were anchored to neutrophils (linear, reticular, membranous, or spot-like structures). NETs seemed to be associated with the formation of platelet aggregates or leukocyte-platelet aggregates. These observations may suggest the adverse effect of intravascular NETs on the host during a sepsis.

Research Products

• [Journal Article] Dynamic pathology for leukocyte-platelet formation in sepsis model. (2015)
  • Author(s): Koike Y, Tanaka K, Kobayashi M, Toiyama Y, Inoue Y, Mohri Y, Uchida K, Mizoguchi A, Kusunoki M.
  • Journal Title: J Surg Res. Volume: 195 Issue: 1 Pages: 188-195
  • DOI: 10.1016/j.jss.2014.05.016
  • Peer Reviewed
• [Journal Article] Risk factors for anastomotic leakage and favorable antimicrobial treatment as empirical therapy for intra-abdominal infection in patients undergoing colorectal surgery. (2014)
  • Author(s): Kobayashi M, Mohri Y, Ohi M, Inoue Y, Araki T, Okita Y, Kusunoki M.
  • Journal Title: Surg Today. Volume: 44 Issue: 3 Pages: 487-493
  • DOI: 10.1007/s00595-013-0575-8
  • Peer Reviewed
• [Journal Article] Infantile Crohn's disease is one of the risk factors of Catheter-related Blood Stream Infection. (2014)
  • Author(s): Koike Y, Uchida K, Inoue M, Ide S, Hashimoto K, Matsushita K, Otake K, Ohkita Y, Araki T, Kobayashi M, Mohri Y, Kusunoki M.
  • Journal Title: Pediatr Int. Volume: Feb 12 Issue: 3 Pages: 364-368
  • DOI: 10.1111/ped.12304
  • Peer Reviewed
• [Presentation] 虚血再灌流臓器障害における全身性炎症反応とNeutrophil Extracellular Trapsとの関連 (2015)
  • Author(s): 田中光司、小池勇樹、井出正造、今岡裕基、野口智史、森 浩一郎、沖上正人、川村幹雄、奥川喜永、三枝 晋、問山裕二、大井正貴、小林美奈子、井上靖浩、荒木俊光、内田恵一、毛利靖彦、楠 正人
  • Organizer: 第115回日本外科学会定期学術集会
  • Place of Presentation: 名古屋国際会議場愛知県名古屋市
  • Year and Date: 2015-04-16
• [Presentation] SSI予防対策の標準化 術前腸管処置 (2014)
  • Author(s): 小林 美奈子, 毛利 靖彦, 大北 喜基, 井上 幹大, 荒木 俊光, 田中 光司, 井上 靖浩, 問山 裕二, 大井 正貴, 内田 恵一, 楠 正人
  • Organizer: 第26回日本外科感染症学会総会学術集会
  • Place of Presentation: 神戸国際会議場
• [Presentation] 術後感染予防抗菌薬使用の標準化 術後感染予防抗菌薬の標準的使用と本邦での変遷 (2014)
  • Author(s): 小林 美奈子, 毛利 靖彦, 大北 喜基, 井上 幹大, 荒木 俊光, 田中 光司, 井上 靖浩, 問山 裕二, 大井 正貴, 内田 恵一, 楠 正人
  • Organizer: 第26回日本外科感染症学会総会学術集会
  • Place of Presentation: 神戸国際会議場