プラズマ照射による組織細胞への分子レベルでの影響の超微形態学的解析

• Project Area: Plasma medical innovation
• Project/Area Number: 15H00901
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Kyorin University
• Principal Investigator: 秋元 義弘 杏林大学, 医学部, 教授 (60184115)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000); Fiscal Year 2016: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2015: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
• Keywords: プラズマ医療 / 創傷治癒 / 皮膚 / α平滑筋アクチン / O-GlcNAc修飾 / 病原性原虫 / トリパノソーマ / 免疫組織化学 / プラズマ / ガレクチン

Outline of Annual Research Achievements

今年度はまず皮膚の創傷治癒過程におけるプラズマ照射による細胞骨格タンパク質の糖修飾 (O-GlcNAc修飾) への影響について検討した。細胞内タンパク質に対する放射線照射の作用には、糖修飾 (O-GlcNAc修飾) を増加させることが知られている。このO-GlcNAc修飾に注目し、プラズマ照射に引き続いて生じる「細胞内タンパク質の糖修飾への影響」を組織化学的に検討して明らかにすることを目的にした。細胞骨格タンパク質のうち特に創傷治癒の過程で重要な働きをする筋線維芽細胞のα平滑筋アクチンに注目して解析を行った。その結果、高周波電気凝固による創傷皮膚ではα平滑筋アクチンはほとんど発現しないのに対して、プラズマ照射した皮膚では発現することが明らかになった。さらにアクチンにおけるO-GlcNAc修飾部位を調べ、そのうちアクチンの機能に重要な部位を選んで抗O-GlcNAc修飾ペプチド抗体を作製した。この抗体を用いてプラズマ照射した創傷皮膚におけるO-GlcNAc修飾アクチンの局在を免疫組織化学法により検討した結果、O-GlcNAc修飾アクチンは筋線維芽細胞の核および細胞質に局在することが明らかになった。
次にプラズマ照射によるドナー血液からの病原性原虫の除去に関する超微形態学的検証を行った。ヒトや動物の血液に寄生し、睡眠病、貧血や発熱を引き起こすトリパノソーマ原虫（Trypanosoma brucei）を研究対象とした。トリパノソーマ原虫は独特な細胞内小器官を持っており、それらの超微細形態変化を走査電子顕微鏡と透過型電子顕微鏡で解析した。その結果、プラズマ照射により原虫に異常なねじれと膨化が生じ、さらに細胞膜が崩壊して死滅していく様子が観察された。その際、プラズマ照射によりミトコンドリアや小胞体の膨化が起こることが明らかになった。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] The Johns Hopkins School of Medicine(米国)
• [Journal Article] Galectin expression in healing wounded skin treated with low-temperature plasma：Comparison with treatment by electronical coagulation. (2016)
  • Author(s): Akimoto Y, Ikehara S, Yamaguchi T, Kim J, Hayato Kawakami, Nobuyuki Shimizu, Masaru Hori, Hajime Sakakita, Ikehara Y.
  • Journal Title: Archives of Biochemistry and Biophysics Volume: 印刷中 Pages: 86-94
  • DOI: 10.1016/j.abb.2016.01.012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Red Blood Cell Coagulation Induced by Low-temperature Plasma Treatment. (2016)
  • Author(s): Miyamoto K, Ikehara S, Takei H, Akimoto Y, Sakakita H, Ishikawa K, Ueda M, Ikeda J, Yamagishi M, Kim J, Yamaguchi T, Nakanishi H, Shimizu N, Hori M, Ikehara Y.
  • Journal Title: Arch Biochem Biophys Volume: in press Pages: 95-101
  • DOI: 10.1016/j.abb.2016.03.023
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Mucin-type core 1 glycans regulate the localization of neuromuscular junctions and establishment of muscle cell architecture in Drosophila. (2016)
  • Author(s): K.Itoh, Y.Akimoto, TJ.Fuwa, C.Sato, A.Komatsu, S.Nishihara.
  • Journal Title: Dev Biol Volume: 412 Issue: 1 Pages: 114-127
  • DOI: 10.1016/j.ydbio.2016.01.032
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] VAMP7 regulates autophagy to maintain mitochondrial homeostasis and to control insulin secretion in pancreatic beta-cells. (2016)
  • Author(s): Aoyagi K, Ohara-Imaizumi M, Itakura M, Torii S, Akimoto Y, Nishiwaki C, Nakamichi Y, Kishimoto T, Kawakami H, Harada A, Takahashi M, Nagamatsu S
  • Journal Title: Diabetes Volume: 65 Issue: 6 Pages: 1648-1659
  • DOI: 10.2337/db15-1207
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A novel podocyte gene, Semaphorin 3G, protects glomerular podocyte from lipopolysaccharide-induced inflammation. (2016)
  • Author(s): Ishibashi R, Takemoto M, Akimoto Y, Ishikawa T, He P, Maezawa Y, Sakamoto K, Tsurutani Y, Ide S, Ide K, Kawamura H, Kobayashi K, Tokuyama H, Tryggvason K, Betsholtz C, Yokote K
  • Journal Title: Scientific Report Volume: 6 Issue: 1 Pages: 25955-25955
  • DOI: 10.1038/srep25955
  • Peer Reviewed / Open Access
• [Journal Article] Pre-embedding method of electron microscopy for glycan localization in mammalian tissues and cells using lectin probes. (2016)
  • Author(s): Akimoto Y, Takata K, Kawakami H
  • Journal Title: Methods Mol. Biol. Volume: 1474 Pages: 259-267
  • DOI: 10.1007/978-1-4939-6352-2_16
  • ISBN: 9781493963508, 9781493963522
  • Peer Reviewed
• [Journal Article] 糖鎖の局在を見る (2016)
  • Author(s): 秋元義弘、遠藤玉夫、川上速人
  • Journal Title: Clinical Neuroscience Volume: 34 Pages: 643-645
• [Journal Article] Fluorescent imaging of endothelial glycocalyx layer with wheat germ agglutinin using intravital microscopy. (2016)
  • Author(s): Kataoka H, Ushiyama A, Kawakami H, Akimoto Y, Matsubara S, Iijima T
  • Journal Title: Microsc. Res. Tech. Volume: 79 Issue: 1 Pages: 31-37
  • DOI: 10.1002/jemt.22602
  • Peer Reviewed
• [Journal Article] Plasma blood coagulation without involving the activation of platelets and coagulation factors. (2015)
  • Author(s): Ikehara S, Sakakita H, Ishikawa K, Akimoto Y, Yamaguchi T, Yamagishi M, Kim J, Ueda M, Ikeda J, Nakanishi H, Shimizu N, Hori M, Ikehara Y
  • Journal Title: Plasma Process. Polym. Volume: 12 Issue: 12 Pages: 1348-1353
  • DOI: 10.1002/ppap.201570045
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Enrichment of O-GlcNAc-modified peptides using novel thiol-alkyne and thiol-disulfide exchange. (2015)
  • Author(s): Tsumoto H, Ogasawara D, Hashii N, Suzuki T, Akimoto Y, Endo T, Miura Y
  • Journal Title: Bioorg. Med. Chem. Lett. Volume: 25 Issue: 13 Pages: 2645-2649
  • DOI: 10.1016/j.bmcl.2015.04.082
  • Peer Reviewed
• [Journal Article] Sentinel node biopsy for the head and neck using contrast-enhanced ultrasonography combined with indocyanine green fluorescence in animal models: a feasibility study. (2015)
  • Author(s): Kogashiwa Y, Sakurai H, Akimoto Y, Sato D, Ikeda T, Matsumoto Y, Moro Y, Kimura T, Hamanoue Y, Nakamura T, Yamauchi K, Saito K, Sugasawa M, Kohno N
  • Journal Title: PLoS ONE Volume: 10 Issue: 7 Pages: e0132511-e0132511
  • DOI: 10.1371/journal.pone.0132511
  • Peer Reviewed / Open Access
• [Presentation] Molecular morphological study of healing wounded skin treated with low-temperature plasma: comparison with treatment by electronical coagulation. (2017)
  • Author(s): Akimoto Y, Ikehara S, Yamaguchi T, Kim J, Kawakami H, Shimizu N, Hori M, Sakakita H, Ikehara Y
  • Organizer: International Conference on Plasma Medical Science Innovation (ICPMSI) 2017
  • Place of Presentation: Nagoya
  • Year and Date: 2017-02-26
  • Int'l Joint Research
• [Presentation] Immunoelectron microscopic study of healing wounded skin treated with low-temperature plasma. (2016)
  • Author(s): Akimoto Y, Ikehara S, Yamaguchi T, Kim J, Kawakami H, Shimizu N, Hori M, Sakakita H, Ikehara Y
  • Organizer: 日本顕微鏡学会第59回シンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2016-11-18
• [Presentation] Plasma-treatment inhibits the in vitro growth of Trypanosoma brucei. (2016)
  • Author(s): Yokoyama N, Sakakida H, Akimoto Y, Sivakumar T, Inoue N, Ishikawa K, Hori M, Ikehara Y
  • Organizer: 第57回日本熱帯医学会大会
  • Place of Presentation: 東京
  • Year and Date: 2016-11-05
• [Presentation] Molecular morphological approach for the mechanism of wound healing with non-thermal plasma. (2016)
  • Author(s): Akimoto Y
  • Organizer: Workshop “Current concept for LTP at atmospheric pressure in medical practice”
  • Place of Presentation: Munich, Germany
  • Year and Date: 2016-09-11
• [Presentation] Efficacy of intraperitoneal therapy with plasma-activated medium targeting peritoneal micrometastasis as revealed by human gastric cancer cell lines in nude mice. (2016)
  • Author(s): Nakanishi H, Tanaka H, Akimoto Y, Tanaka A, Hori M, Ikehara Y
  • Organizer: 6th International Conference on Plasma Medicine
  • Place of Presentation: Bratislava, Slovakia
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Mechanism of blood coagulation induced by plasma treatment in addition to the acceleration of the natural blood coagulation process. (2016)
  • Author(s): Ikehara Y, Ikehara S, Akimoto Y, Sakakita H, Ishikawa K, Ueda M, Ikeda J, Kim J, Yamaguchi T, Nakanishi H, Shimizu T, Shimizu N, Hori M
  • Organizer: 6th International Conference on Plasma Medicine
  • Place of Presentation: Bratislava, Slovakia
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Effect of plasma-treatment on the in vitro growth of Trypanosoma brucei. (2016)
  • Author(s): Yokoyama N, Sakakida H, Akimoto Y, Sivakumar T, Ishikawa K, Hori M, Ikehara Y
  • Organizer: 6th International Conference on Plasma Medicine
  • Place of Presentation: Bratislava, Slovakia
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Expression of galectins and alpha smooth muscle actin in low-temperature plasma-treated healing skin. (2016)
  • Author(s): Akimoto Y, Ikehara S, Yamaguchi T, Kim J, Kawakami H, Shimizu N, Hori M, Sakakita H, Ikehara Y
  • Organizer: 6th International Conference on Plasma Medicine
  • Place of Presentation: Bratislava, Slovakia
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] Health effects of repeated intraperitoneal injections of plasma-activated medium in mice. (2016)
  • Author(s): Tanaka A, Hirata M, Ikehara Y, Akimoto Y, Nakanish H, Tanaka H, Hori M
  • Organizer: 6th International Conference on Plasma Medicine
  • Place of Presentation: Bratislava, Slovakia
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] シンポジウム「プラズマ医療科学の創成にむけて」プラズマ照射による創傷治癒への影響の組織細胞化学的解析. (2016)
  • Author(s): 秋元義弘
  • Organizer: 第57 回日本組織細胞化学会総会・学術集会
  • Place of Presentation: 杏林大学井の頭キャンパス、三鷹市、東京
  • Year and Date: 2016-09-03
• [Presentation] Plasma treatment induces blood clot formation; protein aggregation and hemolysis. (2016)
  • Author(s): 1.Ikehara Y, Ikehara S, Yamaguchi T, Akimoto Y, Sakakita H, Kim J, Ishikawa K, Hori M, Nakanishi H, Shimizu N
  • Organizer: 43rd IEEE International Conference on Plasma Science
  • Place of Presentation: Banff, Canada
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] Increase in galectin expression in healing wounded skin treated with low-temperature plasma: comparison with treatment by electronical coagulation. (2016)
  • Author(s): Akimoto Y, Ikehara S, Yamaguchi T, Kim J, Kawakami H, Shimizu N, Hori M, Sakakita H, Ikehara Y
  • Organizer: 43rd IEEE International Conference on Plasma Science
  • Place of Presentation: Banff, Canada
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] 大気圧低温プラズマ照射による皮膚創傷治癒への影響の超微形態学的解析 (2016)
  • Author(s): 1.秋元義弘、池原早苗、山口高志、金 載浩、清水伸幸、堀 勝、榊田 創、松原幸枝、三浦知子、関口純理、川上速人、池原 譲
  • Organizer: 日本顕微鏡学会第72回学術講演会
  • Place of Presentation: 仙台
  • Year and Date: 2016-06-14
• [Presentation] 糖尿病性腎症に伴う糖修飾アクチンの局在の変化. (2016)
  • Author(s): 秋元義弘、三浦ゆり、戸田年総、福冨俊之、菅原大介、Gerald W Hart、楊 國昌、遠藤玉夫、川上速人
  • Organizer: 第121回日本解剖学会総会・全国学術集会
  • Place of Presentation: 郡山
  • Year and Date: 2016-03-28
• [Presentation] 粒子径10nm銀ナノ粒子の安全性評価に関する検討. (2016)
  • Author(s): 小林尚樹、磯田勝広、長谷川知也、秋元義弘、平裕一郎、平郁子、石田功
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-26
• [Presentation] プラズマ照射による細胞，組織への影響の分子形態学的解析. (2015)
  • Author(s): 秋元義弘
  • Organizer: 第４回 プラズマ医療科学の創成研究会
  • Place of Presentation: 神戸
  • Year and Date: 2015-10-30
• [Presentation] 蛍光in situ PLA法と可視光in situ PLA法の比較検討. (2015)
  • Author(s): 秋元義弘、菅原大介、宮東昭彦、川上速人
  • Organizer: 第56回日本組織細胞化学会総会・学術集会
  • Place of Presentation: 枚方
  • Year and Date: 2015-10-03
• [Presentation] The effects on the endothelial glycocalyxlayer and the microcirculatory parameters under septic condition in mice. (2015)
  • Author(s): Kataoka H, Ushiyama A, Kawakami H, Akimoto Y, Matsubara S, Ochi H, Iijima T
  • Organizer: The 10th world congress for microcirculation
  • Place of Presentation: Kyoto
  • Year and Date: 2015-09-25
  • Int'l Joint Research
• [Presentation] Effect of low temperature plasma on the expression of galectins in wound healing skin. (2015)
  • Author(s): Akimoto Y, Sakakita H, Ikehara S, Yamaguchi T, Kim J, Ikehara Y
  • Organizer: 23rd International Symposium on Glycoconjugates
  • Place of Presentation: Split, Croatia
  • Year and Date: 2015-09-15
  • Int'l Joint Research
• [Presentation] プラズマ照射による組織細胞への分子レベルでの影響の超微形態学的解析. (2015)
  • Author(s): 秋元義弘、榊田 創、池原早苗、山口高志、金 載浩、堀 勝、池原 譲
  • Organizer: 第４回プラズマ医療の創成公開シンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2015-08-05
• [Presentation] レチノイン酸によるラット胎仔表皮の粘膜上皮への分化転換. (2015)
  • Author(s): 秋元義弘、帯刀章子、宮地まり、鎌田理代、松原幸枝、川上速人
  • Organizer: 日本顕微鏡学会第71回学術講演会
  • Place of Presentation: 京都
  • Year and Date: 2015-05-13
• [Book] Diabetes and O-GlcNAcylation. In Glycoscience: Biology and Medicine. (Taniguchi N, Endo T, Hart GW, Seeberger PH, Wong C-H, Eds.) (2015)
  • Author(s): Akimoto Y, Miura Y, Endo T, Kawakami H, Hart GW
  • Total Pages: 1557
  • Publisher: Springer
• [Book] 糖鎖の新機能開発・応用ハンドブック～創薬・医療から食品開発まで (2015)
  • Author(s): 秋元義弘、川上速人
  • Total Pages: 678
  • Publisher: ヌ・ティー・エス
• [Remarks] 杏林大学医学部解剖学教室 顕微解剖学部門 研究テーマ
  • URL: http://www.kyorin-u.ac.jp/univ/user/medicine/anat2/index.php/%E7%A0%94%E7%A9%B6/%E7%A0%94%E7%A9%B6%E3%83%86%E3%83%BC%E3%83%9E
• [Remarks] 杏林大学医学部解剖学教室 顕微解剖学部門
  • URL: http://www.kyorin-u.ac.jp/univ/user/medicine/anat2/