健康科学研究所::研究室紹介::橋田 誠一

生活習慣病部門 橋田 誠一

2017/03/22
学位
医学博士 徳島大学

専門分野
生化学、臨床化学、免疫化学

職  歴
1976年 徳島大学医学部栄養学科卒業
1978年 徳島大学大学院栄養学研究科修士課程修了
1982年 徳島大学大学院医学研究科博士課程修了
1982年 宮崎医科大学助手
1984年 宮崎医科大学講師
1987年 宮崎医学大学助教授
1991年 University of California Davis(USA, California)客員研究員
2003年 宮崎大学医学部助教授(統合)
2004年 徳島文理大学人間生活学部食物栄養学科教授、
徳島文理大学健康科学研究所免疫アレルギー部門教授併任
2010年 徳島文理大学健康科学研究所生活習慣病部門教授併任

学  会
日本生化学会、日本内分泌学会、American Society for Microbiology
日本糖尿病学会、日本臨床化学学会、日本肥満学会、日本栄養食糧学会、日本栄養改善学会

研究の概要
高分子抗原およびペプチド抗原の高感度酵素免疫測定法や抗体の高感度検出法を開発し、その生理機能の解析や病態との関連さらに診断への応用を行っています。また最近では,糖尿病予防教室を主催し、非侵襲性試料中のバイオマーカーを基盤とした運動・栄養指導法の開発を行っています。
1)タンパク質やペプチドの超高感度免疫測定法の開発とそれを応用した臨床診断法の開発を行っています。新しい抗体の酵素標識法の開発や新しい測定システムを開発する事により、従来の測定法では検出されなかった超微量のタンパク質やペプチドの測定を可能としてきました。その結果、今まで複雑な過程を必要としていた微量抗原の直接測定が可能となり、広範な測定対象を可能としました。現在では、検出感度はzeptomole(1x10-21mol; 約600分子)に達しています。これらの高感度測定法により、尿中ヒト成長ホルモン(hGH)の直接の測定を可能とし、尿による小人症の診断を可能としました。また、血中hTSH、hLH測定により、バセドウ病、性的発達不足の診断を可能としました。さらに超高感度化することにより、RT-PCRによるHIV-1 RNA検出と同時期にp24抗原を検出する事が可能となり、早期診断に貢献しました。最近では、新しい活性ペプチド・アドレノメジュリンやPAMPさらにグレリンを抽出・濃縮操作なしに直接に、しかも活性基を含むインタクト・ペプチドとして初めて測定を可能とし、生理機能や病態の解析に寄与しています。最近では、より的確な心不全の診断法を目指し、ネイティブproBNPやNT-proBNPおよびBNPの高感度分別測定法の開発を行っています。
2)感染や自己免疫疾患の早期診断を目的として抗体の超高感度免疫測定法の開発も行っています。抗体の新しい測定システムを開発する事により、ウエスタンブロット法の1万倍高感度な抗体の検出が可能となりました。その結果、HIV-1 感染の早期診断(平均2週間)を可能としました。さらに抗HIV-1 抗体の超高感度測定法の開発により抗原やRNA検出以前に抗体を検出し、HIV-1 RNAより早期の感染診断の可能性を示しました。また、サイログロブリン自己抗体検出では、バセドウ病や橋本病ばかりでなく全ての健常人血中に自己抗体を検出し、自己免疫疾患の新しい考え方を提唱しました。さらに、ガンの早期診断を目的としてガン自己抗体の超高感度免疫測定法の開発を行っています。
最近では、1型糖尿病やSPIDDMを高精度かつ早期に検出するために、膵島関連自己抗体(インスリン自己抗体、GAD抗体、IA-2抗体等)を高感度に検出する検出法を開発しています。さらに、単独の抗体検出では見逃すことがあるため、新規の高感度マルチ抗体検出法(インスリン自己抗体、GAD抗体、IA-2の同時検出)を開発し、コホート試料を用い、検証しています;地域イノベーション戦略支援プログラム(国際競争力強化地域)「とくしま「健幸」イノベーション構想推進地域」;「膵β細胞をまもる研究:膵島関連自己抗体の高感度検出による膵障害機構の解析と判定への応用」。
3)糖尿病予備群を捕捉するための、非侵襲性試料(尿)中の糖尿病リスク・バイオマーカー(アディポネクチン、レジスチン、レプチン、遊離インスリン受容体、インスリン、TNF-αなど)の超高感度測定法を開発しました。そして、このバイオマーカーを指標とする生活改善指導を行い、糖尿病発症リスクからの回避のための「糖尿病予防教室」を主催しています。
4)糖尿病性腎症に焦点を絞り、尿中アディポネクチンの分子パターンを識別する検出法を開発し、早期腎障害の検出から重症化に至る過程の診断法を開発している;地域イノベーション戦略支援プログラム(国際競争力強化地域)「とくしま「健幸」イノベーション構想推進地域」。

主要論文・著書
1. Hashida, S., Towatari,T., Kominami, E. and Katunuma, N. : Inhibition by E-64 derivatives of rat liver cathepsin B and cathepsin L in vitro and in vivo. J. Biochem. 88 : 1805-1811 (1980)
2. Hashida, S., Nakagawa, K., Yoshitake, S., Imagawa, M., Ishikawa, E., Endo, Y., Ohtaki, S., Ichioka, Y. and Nakajima, K. : A highly sensitive sandwich enzyme immunoassay of human growth hormone in serum using affinity-purified anti-human growth hormone Fab'-horseradish peroxidase conjugate. Anal. Lett. 16 (B1) : 31-44 (1983)
3. Hashida, S., Nakagawa, K., Imagawa, M., Inoue, S., Yoshitake, S., Ishikawa, E., Endo, Y., Ohtaki, S., Ichioka, Y. and Nakajima, K. : Use of inorganic salts to minimize serum interference in a sandwich enzyme immunoassay for human growth hormone using Fab'-horseradish peroxidase conjugate. Clin. Chim. Acta 135 : 263-273 (1983)
4. Hashida, S., Imagawa, M., Inoue, S., Ruan, K.H. and Ishikawa, E. : More useful maleimide compounds for the conjugation of Fab' to horseradish peroxidase through thiol groups in the hinge. J. Appl. Biochem. 6 : 56-63 (1984)
5. Hashida, S., Imagawa, M., Ishikawa, E. and Freytag, J.W. : A simple method for the conjugation of affinity-purified Fab' to horseradish peroxidase and ß-D-galactosidase from Escherichia coli. J. Immunoassay 6 : 111-123 (1985)
6. Hashida, S. and Ishikawa, E. : Use of normal IgG and its fragments to lower the non-specific binding of Fab'-enzyme conjugates in sandwich enzyme immunoassay. Anal. Lett. 18 (B9) : 1143-1155 (1985)
7. Hashida, S., Nakagawa, K., Ishikawa, E. and Ohtaki, S. : Basal level of human growth hormone (hGH) in normal serum. Clin. Chim. Acta 151 : 185-186, l985.
8. Hashida, S., Ishikawa, E., Nakagawa, K., Ohtaki, S., Ichioka, T. and Nakajima, K. : Demonstration of human growth hormone in normal urine by a highly specific and sensitive sandwich enzyme immunoassay. Anal. Lett. 18 (B13) : 1623-1634 (1985)
9. Hashida, S., Inoue, S., Ishikawa, E., Mori, T., Imura, H., Ogawa, H., Ichioka, T. and Nakajima, K. : Highly specific and sensitive sandwich enzyme immunoassay for human thyroid-stimulating hormone (hTSH) in serum using monoclonal anti-hTSH ß-subunit IgG1-coated polystyrene balls and polyclonal anti-hTSH ß -subunit Fab'-horseradish peroxidase conjugate. Anal. Lett. 19 (9 & 10) : 1121-1136 (1986)
10. Hashida, S., Ishikawa, E., Kato, Y., Imura, H., Mohri, Z. and Nurakami, Y. : Human growth hormone (hGH) in urine and its correlation to serum hGH examined by a highly sensitive sandwich enzyme immunoassay. Clin. Chim. Acta 162 : 229-235 (1987)
11. Hashida, S., Ishikawa, E., Umehara, H., Kumagai, S., Imura, H., Ohmoto, Y. and Hirai, Y. : Sensitive sandwich enzyme immunoassay of human interleukin-2 produced in vitro by peripheral blood mononuclear cells. Anal. Lett. 20 (7) : 1083-1097 (1987)
12. Hashida, S., Tanaka, K., Kohno, T., Ishikawa, E., Umehashi, H., Hayashida, T., Mori, T., Imura, H. and Ogawa, H. : A highly sensitive sandwich enzyme immunoassay of human thyroid-stimulating hormone in dried blood on filter paper discs for mass-screening of neonatal hypothyroidism. J. Clin. Lab. Anal. 1 : 198-204 (1987)
13. Hashida, S., Ishikawa, E., Mohri, Z., Nakanishi, T., Noguchi, H. and Nurakami, Y.: Sensitive sandwich enzyme immunoassay of human growth hormone (hGH) in serum and urine using monoclonal antibody-coated polystyrene balls. Endocrinol. Japon. 35 : 171-180 (1988)
14. Hashida, S., Ishikawa, E., Mohri, Z., Nurakami, Y., Umehashi, H. and Hayashida, T.: Sensitive enzyme immunoassay of human growth hormone (hGH) in dried blood on filter paper discs. Clin. Chem. Enzym. Comms. 1: 39-44 (1988)
15. Hattori, N., Kato, Y., Nurakami, Y., Hashida, S., Ishikawa, E., Mohri, Z. and Imura, H. : Urinary growth hormone levels measured by ultrasensitive enzyme immunoassay in patients with renal insufficiency. J. Clin. Endocrinol. Metab. 66: 727-732 (1988)
16. Hashida, S., Ishikawa, E., Nakao, K., Mukoyama, M. and Imura, H. : Enzyme immunoassay for a-human atrial natriuretic polypeptide _____ direct measurement of plasma level. Clin. Chim. Acta 175 : 11-18 (1988)
17. Hashida, S., Tanaka, K., Kohno, T. and Ishikawa, E. : Novel and ultrasensitive sandwich enzyme immunoassay (sandwich transfer enzyme immunoassay) for antigens. Anal. Lett. 21 : 1141-1154 (1988)
Yano, K., Hashida, S., Okuno, A. and Ishikawa, E. : Serum concentration of chorionic gonadotropin-like substance and luteinizing hormone in children measured by sensitive enzyme immunoassay.
18. Hasui, Y., Suzumiya, J., Marutsuka, K., Sumiyoshi, A., Hashida, S. and Ishikawa, E. : Comparative study of plasminogen activators in cancers and normal mucosae of human urinary bladder. Cancer Res. 49 : 1067-1070 (1989)
19. Tanaka, K., Hashida, S., Kohno, T., Yamaguchi, K. and Ishikawa, E. : Novel and sensitive noncompetitive enzyme immunoassay for peptides. Biochem. Biophys. Res. Commun. 160 : 40-45 (1989)
20. Hashida, S. and Ishikawa, E. : Detection of one milliattomole of ferritin by novel and ultrasensitive enzyme immunoassay. J. Biochem. 108 : 960-964 (1990)
21. Yone, K., Hashida, S., Tanaka, K., Ichikawa, Y. and Ishikawa, E. : Specific and sensitive sandwich enzyme immunoassay for human tumor necrosis factor-alpha. Clin. Chem. Enzym. Comms. 3 : 1-8 (1990)
21. Hashida, S., Tanaka, K., Yamamoto, N., Uno, T., Yamaguchi, K. and Ishikawa, E. : Detection of one attomole of [Arg8]-vasopressin by novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay). J. Biochem. 110 : 486-492 (1991)
22. Hashida, S., Tanaka, K., Yamamoto, N., Uno, T., Yamaguchi, K. and Ishikawa, E. : Novel and sensitive noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for arginine vasopressin in plasma. Anal. Lett. 24 : 1109-1123 (1991)
23. Hashida, S. and Ishikawa, E. : Novel and ultrasensitive noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay) for a-human atrial natriuretic peptide (a-hANP). J. Clin. Lab. Anal. 6 : 201-208 (1992)
24. Goldstein, M.A., Takagi, M., Hashida, S., Shoseyov, O., Doi, R.H. and Segel I.H. : Characterization of the cellulose-binding domain of the clostridium cellulovorans cellulose-binding protein A. J. Bacteriol. 175 : 5762-5768 (1993)
25. Takagi, M., Hashida, S., Goldstein, M.A. and Doi, R.H. : The hydrophobic repeated domain of the clostridium cellulovorans cellulose-binding protein (CbpA) has specific interactions with endoglucanases. J. Bacteriol. 175 : 7119-7122 (1993)
26. Hashida, S., Hirota, K., Hashinaka, K., Saitoh, A., Nakata, A., Shinagawa, H., Oka, S., Shimada, K., Mimaya, J., Matsushita, S. and Ishikawa, E. : Detection of antibody IgG to HIV-1 in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens for diagnosis of HIV-1 infection. J. Clin. Lab. Anal. 7 : 353-364 (1993)
27. Hashinaka, K., Hashida, S., Saitoh, A., Nakata, A., Shinagawa, H., Oka, S., Shimada, K. and Ishikawa E. : Conjugation of recombinant reverse transcriptase of HIV-1 to ß-D-galactosidase from Escherichia coli for ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of anti-HIV-1 IgG. J. Immunol. Methods 172 : 179-187 (1994)
28. Hashida, S., Hashinaka K. and Ishikawa, E. : Ultrasensitive enzyme immunoassay. In: El-gewely, M.R. ed. Biotechnology Annual Review Volume 1, (1995) pp403-451, Elsevier Science Publishers B.V., Amsterdam..
29. Fujimoto, K., Yasue, H., Hashida, S., Nakao, K., Ishikawa, E. and Miyamoto, E. : Augmented expression of artial myosin light chain 1 in ventricular aneurysms of human : Enzyme immunoassay for atrial myosin light chain 1. Biochem. Biophys. Res. Commun. 207 : 75-79 (1995)
30. Sadamatsu, M., Kato, N., Iida, H., Takahashi, S., Sakaue, K., Takahashi, K., Hashida, S. and Ishikawa, E. : The 24-hour rhythms in plasma growth hormone, prolactin and thyroid stimulating hormone : Effect of sleep deprivation. J. Neuroendocrinol. 7 :597-606 (1995)
31. Hashida, S., Hashinaka, K., Nishikata, I., Oka, S., Shimada, K., Saito, A., Takamizawa, A., Shinagawa, H. and Ishikawa, E. : Shortening of the window period in diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgG to p17 and reverse transcriptase in serum with ultrasensitive enzyme immunoassay. J. Virol. Methods 62 : 43-53 (1996)
32. Hashida, S., Hashinaka, K., Nishikata, I., Oka, S., Shimada, K., Saito, A., Takamizawa, A., Shinagawa, H. and Ishikawa, E. : Shortening of the window period in diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgG to p17 and reverse transcriptase in serum with ultrasensitive enzyme immunoassay. J. Virol. Methods 62 : 43-53 (1996)
Hashida, S., Ishikawa, S., Hashinaka, K., Nishikata, I., Oka, S., Shimada, K., Saito, A., Takamizawa, A., 33. Shinagawa, H. and Ishikawa, E. : Optimal conditions of immune complex transfer enzyme immunoassays for antibody IgGs to HIV-1 using recombinant p17, p24, and reverse transcriptase as antigens. J. Clin. Lab. Anal. 12 : 98-107 (1998)
34. Katakami, H., Hashida, S., Hidaka, H., Ishikawa, E. and Matsukura, S. : Development and clinical application of a highly sensitive enzyme immunoassay (EIA) for human growth hormone-releasing hormone (hGHRH) in plasma. Endocrine J. 45(Supple) : S67-S70 (1998)
35. Hirota, K., Hashida, S., Ishikawa, E. and Totani, M. : Sensitive enzyme immunoassay for anti-ß-lactoglobulin IgG in serum. Ann. Clin. Biochem. 35 : 649-655 (1998)
36. Hashida, S., Ishikawa, S., Hashinaka, K., Nishikata, I., Oka, S. and Ishikawa, E.: Earlier detection of human immunodeficiency virus type 1 p24 antigen and immunoglobulin G and M antibodies to p17 antigen in seroconversion serum panels by immune complex transfer enzyme immunoassays. Clin. Diag. Lab. Immunol. 7 : 872-881 (2000)
37. Ohnishi, T., Kakimoto, K., Hashida, S., Fujii, M., Hirono, S., Nishiyama, K., Amita, Y., Ishikawa, E., Tsubouchi, H. and Daikuhara, Y. : Development of highly sensitive enzyme-linked immunosorbent assay for hepatocyte growth factor / scatter factor (HGF / SF): determination of HGF / SF in serum and urine from normal human subjects. J. Immunol. Method. 244 : 163-173 (2000)
38. Hashida S., Ishikawa E., Nakamichi N. and Sekino H.: Concentration of egg white lysozyme in serum of healthy subjects after oral administration. Clinical and Experimental Pharmacology and Physiology. 29: 79-83 (2002)
39. Iga, M., Matsuda, Z., Okayama, A., Sugiura, W., Hashida, S., Morishita, K., Nagai, Y. and Tubouchi, H.: Rapid phenotypic assay for immunodeficiency virus type 1 protease using in vitro translation. J. Virolog. Method. 106: 25-37 (2002)
40. Yamaga, J., Hashida, S., Kitamura, K., Tokashiki, M., Aoki. T., Inatsu, H., Ishikawa, N., Kangawa, K., Morishita, K. and Eto, T.: Direct measurement of glycine-extended adrenomedullin in plasma and tissue using an ultrasensitive immune complex transfer enzyme immunoassay in rat. Hypertens. res. 26: S45-53 (2003)
41. Kita T., Kitamura K., Hashida S., Morishita K. and Eto T.: Plasma adrenomedullin is closely correlated with pulse wave velocity in middle-aged and elderly patients. Hypertens. res. 26: 887-893 (2003)
42. Cao YN, Kitamura K, Ito K, Kato J, Hashida S, Morishita K and Eto T.: Glycine-extended adrenomedullin exerts vasodilator effect through amidation in the rat aorta. Regul. Pept. 113:109-14 (2003)
43. 橋田誠一、片上秀喜:グレリンの超高感度免疫測定法. ホルモンと臨床 51: 775-787(2003)
44. Hashida S., Kitamura K., Nagatomo Y., Shibata Y., Imamura T., Yamada K., Fujimoto S., Kato J., Morishita K. and Eto T. : Development of an ultra-sensitive enzyme immunoassay for human proadrenomedullin N-terminal 20 peptide (PAMP) and direct measurement of two molecular forms of PAMP in plasma from healthy subjects and patients with cardiovascular disease. Clin. Biochem. 37: 14-21 (2004).
45. Chen L., Hashida s., Kitamura K., Eto T., Kangawa K., Serino R., Kis B., Yamashita H., Ueta Y.: Disassociated Increases of Adrenomedullin in the Rat Cerebrospinal Fluid and Plasma after Salt Loading and Systemic Administration of Lipopolysaccharide. Peptides. 25:609-614 (2004)
46. Kawano K., Tajima N., Hashida S.,Ishikawa E.: Study of blood metabolism and urinary excretion of chymopapain following intradiscal injection using a high-sensitivity enzyme immunoassay. J. Orthopaedic Sci.10: 206-213(2005)
47. Akiyama S., Hobara N., Maruo N., Hashida S., Kitamura K., Eto T. and Kawasaki H.: Adrenomedullin release in the rat mesenteric resistance artery. Peptides. 26: 2222-2230 (2005)
48. Inoue S., Hashida S.: Plasma intact ghrelin levels in obese children. Growth Hormone & IGF Research. 16: S55 (2006)
49. Obata T, Yokota I, Yokoyama K, Okamoto E, Kanezaki Y, Tanaka Y, Maegawa H, Teshigawara K, Hirota F, Yuasa T, Kishi K, Hattori A, Hashida S, Masuda K, Matsumoto M, Matsumoto T, Kashiwagi A, Ebina Y.: Soluble insulin receptor ectodomain is elevated in the plasma of patients with diabetes mellitus.
Diabetes. 56: 2028-2035 (2007)
50. Umehara A, Nishioka M, Obata T, Ebina Y, Shiota H, Hashida S: A novel ultra-sensitive enzyme immunoassay for soluble human insulin receptor ectodomain and its measurement in urine from healthy subjects and patients with diabetes mellitus. Clin Biochem 42, 1468-1475. (2009)

最近5年間の論文・総説
1. Numata S., Umehara A., Katakami H., Inoue S. and Hashida S: Development of a novel and ultra-sensitive enzyme immunoassay for human insulin autoantibodies .Clin Biochem 45: 1086-91 (2012)
2. Hashida S., Miyazawa Y., Hirajima Y., Umehara A, Yamamoto M., Numata S. Development of ultra-sensitive enzyme immunoassay for insulin and application to evaluation of the diabetic risk by urine in the morning. In Eltayb Abuelzein ed. Trends in Immunolabelled and Related Techniues. (2012) pp83-100, In Teck d.o.o, Croatia. (ISBN; 978-953-51-0570-1)
3. Tomoyuki Yuasa, Kikuko Amo, Shuhei Ishikura, Hisao Nagaya, Keiji Uchiyama, Seiichi Hashida, Yousuke Ebina. Development of in vitro model of insulin receptor cleavage induced by high glucose in HepG2 Cells. Biochem Biophys Res Commun, 445: 236-243 (2014)
4. 吉村英悟、影山晴秋、鈴木洋子、金澤真雄、瀬野尾章、石塚典子、荒井勝己、今関信夫、髙橋美砂子、佐藤栄子、塩田清二、橋田誠一、井上修二:視床下部腹内側核(VMH)破壊によるガストリン産生細胞(G細胞)の細胞増殖を伴わないガストリン産生および分泌亢進、日本臨床生理学会雑誌 Vol.44, No.2, 61-67, 2014
5. 橋田誠一、山本真弓、沼田聡 : 「高感度ELISAは、こう開発する」-基礎と応用— 臨床化学 43:121-136(2014)
6. 沼田聡、橋田誠一;高感度ELISA定量法;In: 森山達哉,穐山浩 編: 食物アレルギーの現状と低減化食品素材の開発, (2015) pp101-107、シーエムシー出版、東京。
7. Morikawa M. Naito R. Mita K. Watanabe S. Nakanaishi K. Yoshimura T. Miura T. Hashida S. Ito E. Subattomole detection of adiponectin in urine by ultrasensitive ELISA coupled with thio-NAD cycling. Biophysics and Bhysicobiology, 12:79-86 (2015)
8. Ito E. Kaneda M. Kodama H. Tai M. Aoki K Watanabe S. Nakanishi K. Hashida S. Tada S. Kuroda K. Imachi H. Yamashita M. Yoshimura T. Miura N.: Immunoreactive insulin in diabetes mellitus patient sera detected by ultrasensitive ELISA with thio-NAD cycling. Bio Techniques 59:359-367 (2015)
9. Numata S., Umehara A., Katakami H., Inoue S. and Hashida S: Development of a novel ultra-sensitive enzyme immunoassay for human GAD65 antibody. Annals Clinical Biochemistry, 53:495-503 (2016)
10. Hamada, Junpei, Onuma, Hiroshi, Ochi, Fumihiro, Hirai, Hiroki, Takemoto, Koji, Miyoshi, Akiko, Matsushita, Manami, Kadota, Yuko, Ohashi, Jun, Kawamura, Ryoichi, Takata, Yasunori, Nishida, Wataru, Hashida, Seiichi, Ishii, Eiichi, Osawa, Haruhiko. Endoplasmic reticulum stress induced by tunicamycin increases resistin mRNA through the PERK-ATF4-CHOP pathway in THP-1 human monocytes. Journal of Diabetes Investigation 7: 312–323 (2016) 
11. Yuasa T, Amo-Shiinoki K, Ishikura S, Takahara M, Matsuoka T, Kaneto H, Kuroda A, Matsuhisa M, Hashida S.: Sequential cleavage of insulin receptor by calpain 2 and γ-secretase impairs insulin signalling. Diabetologia.59:2711-2721 (2016).
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