Elucidation of molecular mechanism for development of atherosclerosis by cytotoxic factors in cigarette smoke

• Project/Area Number: 26860166
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General pharmacology
• Research Institution: Hokkaido University
• Principal Investigator: Higashi Tsunehito 北海道大学, 医学研究科, 助教 (90453018)
• Research Collaborator: MAI Yosuke
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: タバコ煙ガス相 / 不飽和カルボニル化合物 / 細胞傷害 / プロテインキナーゼC / 細胞毒性 / 血管平滑筋細胞 / タバコ煙抽出物 / 抗酸化物質

Outline of Final Research Achievements

Cigarette smoking is one of the risk factors for cardiovascular diseases. The objective of this study is to elucidate the molecular mechanism(s) for cell injury induced by unsaturated carbonyl compounds in the gas phase of cigarette smoke. First, the method for preparation of gas phase extract of cigarette smoke was optimized. The cell injury caused by both gas phase extract of cigarette smoke and unsaturated carbonyl compounds was intracellular calcium-dependent. They also activated protein kinase C in intracellular calcium-dependent manner. Several antioxidants such as reduced form of glutathione and vitamin E suppressed cell injury induced by unsaturated carbonyl compounds.

Research Products

• [Journal Article] The dynamics of mucosal-associated invariant T cells in multiple sclerosis (2016)
  • Author(s): Chie Sugimoto, Makoto Hirotan, Kazunori Yoshikiyo, Uichi Koshimizu, Rika Wakao, Takahiro Horinouchi, Yuichi Mazaki, Tsunehiko Higashi, Toshiyuki Fukazawa, Hiroyoshi Fujita, Hidenao Sasaki and Hiroshi Wakao
  • Journal Title: SpringerPlus Volume: 5 Issue: 1 Pages: 1259-1259
  • DOI: 10.1186/s40064-016-2923-9
  • NAID: 120005906931
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Standardized Method for the Preparation of a Gas Phase Extract of Cigarette Smoke (2016)
  • Author(s): Higashi T, Mai Y, Mazaki Y, Horinouchi T, Miwa S
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 39 Issue: 6 Pages: 898-902
  • DOI: 10.1248/bpb.b16-00062
  • NAID: 130005155132
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Carbonyl Compounds in the Gas Phase of Cigarette Mainstream Smoke and Their Pharmacological Properties (2016)
  • Author(s): Horinouchi T, Higashi T, Mazaki Y, Miwa S
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 39 Issue: 6 Pages: 909-914
  • DOI: 10.1248/bpb.b16-00025
  • NAID: 130005155133
  • ISSN: 0918-6158, 1347-5215
  • Peer Reviewed
• [Journal Article] Endothelin-1 suppresses insulin-stimulated Akt phosphorylation and glucose uptake via GPCR kinase 2 in skeletal muscle cells (2016)
  • Author(s): Horinouchi T, Hoshi A, Harada T, Higa T, Karki S, Terada T, Higashi T, Mai Y, Nepal P, Mazaki Y, Miwa S
  • Journal Title: Br J Pharmacol Volume: 173 Issue: 6 Pages: 1018-1032
  • DOI: 10.1111/bph.13406
  • NAID: 120005981358
  • Peer Reviewed
• [Journal Article] Using Phos-Tag in western blotting analysis to evaluate protein phosphorylation (2016)
  • Author(s): Horinouchi T, Terada K, Higashi T, Miwa S
  • Journal Title: Methods Mol Biol Volume: 1397 Pages: 267-277
  • DOI: 10.1007/978-1-4939-3353-2_18
  • ISBN: 9781493933518, 9781493933532
• [Journal Article] A standard method for preparation of gas phase extract of cigarette smoke (2016)
  • Author(s): Higashi T, Mai Y, Mazaki Y, Horinouchi T, Miwa S
  • Journal Title: Bull Pharm Biol Volume: in press
  • NAID: 130005155132
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Decreased proteosomal function accelerates cigarette smoke-induced pulmonary emphysema in mice. (2015)
  • Author(s): Yamada Y, Tomaru U, Ishizu A, Ito T, Kiuchi T, Ono A, Miyajima S, Nagai K, Higashi T, Matsuno Y, Dosaka-Akita H, Nishimura M, Miwa S, Kasahara M
  • Journal Title: Lab Invest Volume: 95 Issue: 6 Pages: 625-634
  • DOI: 10.1038/labinvest.2015.43
  • Peer Reviewed
• [Journal Article] Ubiquitination-regulated receptor trafficking of endothelin type A and type B receptors (2015)
  • Author(s): 寺田晃士、堀之内孝広、東恒仁、Prabha Nepal、三輪聡一
  • Journal Title: Folia Pharmacologica Japonica Volume: 145 Issue: 1 Pages: 4-9
  • DOI: 10.1254/fpj.145.4
  • NAID: 130004827712
  • ISSN: 0015-5691, 1347-8397
• [Journal Article] Endothelin-1 activates extracellular signal-regulated kinases 1/2 via transactivation of platelet-derived growth factor receptor in rat L6 myoblasts. (2014)
  • Author(s): Harada T, Horinouchi T, Higa T, Hoshi A, Higashi T, Terada K, Mai Y, Nepal P, Horiguchi M, Hatate C, Miwa S.
  • Journal Title: Life Sci. Volume: 104 Issue: 1-2 Pages: 24-31
  • DOI: 10.1016/j.lfs.2014.04.002
  • Peer Reviewed
• [Journal Article] A simple and rapid method for standard preparation of gas phase extract of cigarette smoke. (2014)
  • Author(s): Higashi T., Mai Y., Noya Y., Horinouchi T., Terada K., Hoshi A., Nepal P., Harada T., Horiguchi M., Hatate C., Kuge Y. and Miwa S.
  • Journal Title: PLos ONE Volume: 9 Issue: 9 Pages: e107856-e107856
  • DOI: 10.1371/journal.pone.0107856
  • NAID: 120005496512
  • Peer Reviewed / Open Access
• [Journal Article] Agonist-promoted ubiquitination differentially regulates receptor trafficking of endothelin type A and type B receptors. (2014)
  • Author(s): Terada K., Horinouchi T., Fujioka Y., Higashi T., Nepal P., Horiguchi M., Sarita K., Hatate C., Hoshi A., Harada T., Mai Y., Ohba Y. and Miwa S.
  • Journal Title: J. Biol. Chem. Volume: 289 Issue: 51 Pages: 35283-35295
  • DOI: 10.1074/jbc.m113.544171
  • NAID: 120005549588
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Unsaturated carbonyl compounds in the gas phase extract of cigarette smoke induce cell death through intracellular Ca2+-dependent PKC activation (2016)
  • Author(s): Higashi T, Mai Y, Mazaki Y, Horinouchi T, Miwa S
  • Organizer: American Society for Cell Biology 2016 Annual Meeting
  • Place of Presentation: San Francisco (USA)
  • Year and Date: 2016-12-03
  • Int'l Joint Research
• [Presentation] タバコ煙ガス相による細胞傷害の抑制因子の探索と抑制メカニズムの解明 (2016)
  • Author(s): 東恒仁、眞井洋輔、堀之内孝広、真崎雄一、三輪聡一
  • Organizer: 第89回日本薬理学会年会
  • Place of Presentation: パシフィコ横浜会議センター（神奈川県横浜市）
  • Year and Date: 2016-03-09
• [Presentation] タバコ煙に起因する細胞傷害の抑制物質の探索と抑制メカニズムの解明 (2015)
  • Author(s): 東恒仁、眞井洋輔、堀之内孝広、真崎雄一、堀口美香、三輪聡一
  • Organizer: 第25回日本循環薬理学会
  • Place of Presentation: 東大寺総合文化センター 金鐘ホール（奈良県奈良市）
  • Year and Date: 2015-12-04
• [Presentation] タバコ煙に起因する細胞傷害の抑制方法の開発およびその分子機構の解明 (2015)
  • Author(s): 東恒仁、眞井洋輔、堀之内孝広、真崎雄一、堀口美香、三輪聡一
  • Organizer: 第66回日本薬理学会北部会
  • Place of Presentation: 富山国際会議場（富山県富山市）
  • Year and Date: 2015-09-18
• [Presentation] グルタチオン・N-アセチルシステイン・システインはタバコ煙ガス相抽出物による細胞傷害に対する保護作用を有する (2015)
  • Author(s): 東恒仁、眞井洋輔、Enas Elmeligy、堀之内孝広、寺田晃士、星暁壮、堀口美香、Prabha Nepal、Sarita Karki、旗手千鶴、三輪聡一
  • Organizer: 第88回日本薬理学会年会
  • Place of Presentation: 名古屋国際会議場（愛知県・名古屋市）
  • Year and Date: 2015-03-18 – 2015-03-20
• [Presentation] Gas phase extract of cigarette smoke and its cytotoxic components induce cell death via protein kinase C and NADPH oxidase (2014)
  • Author(s): Higashi T, Mai Y, Horinouchi T, Terada K, Elmeligy E, Hoshi A, Nepal P, Harada T, Karki S, Horiguchi M, Hatate C, Miwa S
  • Organizer: The 2014 ascb/ifcb Meeting
  • Place of Presentation: Philadelphia (United States of America)
  • Year and Date: 2014-12-06 – 2014-12-10
• [Presentation] タバコ煙中の細胞傷害因子の同定とその細胞傷害メカニズムの解明 (2014)
  • Author(s): 東恒仁
  • Organizer: 日本生物物理学会北海道支部会・北海道分子生物学研究会・日本生化学会2014年度合同シンポジウム
  • Place of Presentation: 北海道大学農学部（北海道・札幌市）
  • Year and Date: 2014-11-19
  • Invited
• [Presentation] タバコ煙による細胞傷害の定量的検出方法の確立と細胞傷害抑制物質の探索 (2014)
  • Author(s): 旗手千鶴、東恒仁、Enas Elmeligy、眞井洋輔、堀之内孝広、寺田晃士、星暁壮、Prabha Nepal、Sarita Karki、堀口美香、原田拓弥、三輪聡一
  • Organizer: 第65回日本薬理学会北部会
  • Place of Presentation: コラッセふくしま（福島県・福島市）
  • Year and Date: 2014-09-26
• [Remarks] 北海道大学大学院医学研究院細胞薬理学教室ホームページ
  • URL: http://plaza.umin.ac.jp/cellular_pharm
• [Remarks] 北海道大学大学院医学研究科細胞薬理学分野ホームページ
  • URL: http://saibo-yakuri.med.hokudai.ac.jp/