イシハラ ケンジ石原 研治教授Kenji ISHIHARA
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- 〔主要な業績〕外国人児童生徒等に対する校内連携における教員の意識 -茨城県内の小中学校教員への質問紙調査から-
学校保健研究, 2023年06月20日, ［査読有り］ - 〔主要な業績〕Effects of salicylate derivatives on localization of p.H723R allele product of SLC26A4
Michio Murakoshi; Yuhi Koike; Shin Koyama; Shinichi Usami; Kazusaku Kamiya; Katsuhisa Ikeda; Yoichi Haga; Kohei Tsumoto; Hiroyuki Nakamura; Noriyasu Hirasawa; Kenji Ishihara; Hiroshi Wada, OBJECTIVE: Pendrin is a transmembrane protein encoded by the SLC26A4 gene that functions in maintaining ion concentrations in the endolymph of the inner ear, most likely by acting as a chloride/bicarbonate transporter. Variants in the SLC26A4 gene are responsible for sensorineural hearing loss. Although pendrin localizes to the plasma membrane, we previously identified that 8 missense allele products of SLC26A4 were retained in the intracellular region and lost their anion exchange function. We also found that 10 mM salicylate induced the translocation of 4 out of 8 allele products from the intracellular region to the plasma membrane and restored their anion exchanger activity. However, since 10 mM salicylate exhibits cytotoxicity, the use of chemical compounds with less cell toxicity is needed. In the present study, therefore, salicylate derivatives were used as the chemical compounds and their effects on the p.H723R allele products of SLC26A4 were investigated. METHODS: HEK293 cells were transfected with the cDNA of p.H723R. Cell proliferation, viability and toxicity assays were performed to investigate the response and health of cells in culture after treatment with four types of salicylate derivatives, i.e., 2-hydroxybenzyl alcohol, 2,3-dihydroxybenzoic acid, 2'-hydroxyacetophenone and methyl salicylate. The effects of these salicylate derivatives on the localization of the p.H723R were investigated by immunofluorescence microscopy. RESULTS: The application of 10 mM salicylate showed an increase in cell toxicity and decrease in cell viability, leading to a significant decrease in cell proliferation. In contrast, the application of 1 mM salicylate derivatives did not show any significant increase in cell toxicity and decrease in cell viability, corresponding to a logarithmic increase in cell concentration with an increase in culture time. Immunofluorescence experiments showed that the p.H723R retained in the endoplasmic reticulum (ER). Among the salicylate derivatives applied, 2-hydroxybenzyl alcohol induced the translocation of p.H723R from the ER to the plasma membrane 3 h after its application. CONCLUSION: The results obtained showed that 2-hydroxybenzyl alcohol restored the localization of the p.H723R allele products of SLC26A4 from the ER to the plasma membrane at a concentration of 1 mM by 3 h after its administration with less cytotoxicity than 10 mM salicylate.
Auris Nasus Larynx, 2022年03月16日, ［査読有り］ - 日本再生医療学会総会ジュニアセッションでの主体的・対話的で深い学びをめざす試み
川上 雅弘; 石原 研治,日本再生医療学会では，第15 回総会（平成27 年度）からジュニアセッションを設けている．我々は，2年後の第17 回総会より新たに「中高生のためのセッション」を設定しその企画運営を行なっている．「中高生のためのセッション」ではベーシックコース，アドバンストコース，作文部門を設定し，再生医療だけではなく社会性や協働，そして参加者同士のつながり・研究者とのつながりを趣旨とした．本発表で報告するアドバンストコースは，事前課題「幹細胞／再生医療研究＋○○○○＝□□□□の実現」について，自由に討論してきた○○○○と□□□□に対する回答をチームごとに発表し，1 日を通して参加者全員で各チームの内容を議論・改善し，最後に学会理事を審査員とした発表会の場で発表を行うものである．アンケートから，参加者が知識的側面を学ぶ以外に，コミュニケーション力，表現力や協調性などを学ぶ機会となったことが示唆された．
, 一般社団法人 日本科学教育学会
日本科学教育学会年会論文集, 2020年 - Discovery of (2-aminophenyl)methanol as a new molecular chaperone that rescues the localization of P123S mutant pendrin stably expressed in HEK293 cells
WataruNabeyama. KenjiIshihara. Hyun SeungBan; HiroshiWadad. HiroyukiNakamura, Pendred syndrome is the most common form of syndromic deafness. It is associated with a mutation in the SLC26A4 gene that encodes pendrin, which is thought to maintain the ion concentration of endolymph in the inner ear most likely by acting as a chloride/bicarbonate transporter. Mutations in the SLC26A4 gene are responsible for sensorineural hearing loss. In this study, we established a stable HEK293 cell line expressing P123S mutant pendrin and developed screening methods for compounds that show pharmacological chaperone activity by image analysis using CellInsight (TM). Morphological analysis of stained cells in each well of 96-well plates yielded six compounds in the compound library. Furthermore, fluorescence intensity analysis of the intracellular localization of P123S mutant pendrin in HEK293 cells using FLUOVIE (TM) 5 and cytotoxicity experiments revealed that (2-aminophenyl)methanol 8 is the most promising molecular chaperone to rescue P123S mutant pendrin: the plasma membrane (M)/cytoplasm (C) ratios are 1.5 and 0.9 at the concentrations of 0.3 and 0.1 mM, respectively, and a sustained effect was observed 12 h after removal of the compound from the cell medium. Because the M/C ratio of salicylate, which was previously discovered as a molecular chaperone of P123S mutant pendrin, was approximately 1 at 10 mM concentration and a sustained effect was not observed even at 6 h, (2-aminophenyl)methanol 8 was 100 times more potent and exhibited a longer sustained effect than salicylate. These findings suggest that (2-aminophenyl)methanol 8 is an attractive candidate for therapeutic agent for Pendred syndrome patients. (C) 2017 Elsevier Ltd. All rights reserved., Elsevier
Bioorg Med Chem, 2017年05月01日, ［査読有り］ - Recognition of, interest in, and understanding of induced pluripotent stem cells and regenerative medicine in Japanese students.
Ishihara K; Ichinomiya A; Inami M; Hashimoto T; Yuzawa R; Ishizu M; Hirohara T; Yashiro Y; Takizawa T
Regen Ther., 2016年09月, ［査読有り］ - Ex Vivo Models for the Study of Eosinophils
Steven J. Ackerman; Kenji Ishihara; Noriyasu Hirasawa; Kazuo Ohuchi; Caroline M. Percopo; Sergei I. Ochkur; Kimberly D. Dyer; Lori A. Wagner, Elsevier Inc.
Eosinophils in Health and Disease, 2013年, ［査読有り］ - Induction of thymic stromal lymphopoietin production by xylene and exacerbation of picryl chloride-induced allergic inflammation in mice.
Satou; N; Ishihara; K; Hiratsuka; M; Tanaka; H; Endo; Y; Saito; S; Iwatate; Y; Leonard; W.J; Hirasawa; N, Background: Some chemical compounds in the environment worsen allergic inflammation. In this study, we examined whether organic solvents induce the production of thymic stromal lymphopoietin (TSLP) which elicits Th2-type immune responses. Methods: Organic solvents were painted on the earlobes of BALB/c mice. The expression of TSLP in the ear was determined by ELISA. Results: Xylene and toluene, but not chloroform or ethyl acetate, induced the expression of mRNA for TSLP in the earlobe tissue. Among the aromatic compounds, xylene, especially m-xylene, and trimethylbenzene caused apparent TSLP production. The level of TSLP in the xylene-treated earlobes reached a maximum at 24 h, and TSLP was expressed in epithelial tissues. Production of TSLP was unaffected in mast cell-deficient W/W-v mice but apparently diminished in TNF-alpha knockout mice and IL-4 receptor knockout mice. Repeated painting of xylene for 7 days induced an increase in the weight of cervical lymph nodes and expression of OX40 ligand, both of which were inhibited in TSLP receptor knockout mice. Xylene promoted the picryl chloride-induced thickening of the ear and IL-4 production, which were reversed in TSLP receptor knockout mice. Conclusion: Xylene induced TSLP production, resulting in an exacerbation of allergic inflammation. Thus, xylene might be a good tool for examining the roles of TSLP in eliciting allergy in experimental animals. Copyright (C) 2011 S. Karger AG, Basel, KARGER
Int. Arch. Allergy Immunol., 2012年, ［査読有り］ - Enhancement of nickel elution by lipopolysaccharide-induced inflammation.
Tanaka R; Goi Y; Ishihara K; Ueda K; Narushima T; Ohtsu H; Hiratsuka M; Hirasawa N, Background: Implantations of metallic biomedical devices into bodies are increasing. The elution of Ni ions from these devices can lead to metal allergies. However, the molecular mechanisms of the elution have not been fully examined. Furthermore, it is not clear whether infection and inflammation affect the corrosion of metals.
Objective: We examined whether the elution of Ni from metal wires and plates was enhanced by inflammation in vivo and in vitro.
Methods: A Ni or SUS316L wire was implanted subcutaneously in the dorsum of mice. Lipopolysaccharide (LPS) was injected at the site immediately following the implantation. After 8, 24, and 72 h, the tissue around the wire was excised. RAW 264 cells were seeded on a Ni plate and incubated for 24 h in medium containing LPS. The amount of Ni in the tissue or conditioned medium was determined fluorometrically.
Results: The release of Ni ions from the wire was significantly increased from 8 to 72 h, and further increased by LPS. LPS also enhanced the release of Ni ions by the cells, but only when they were attached to the Ni plate. Chloroquine, bafilomycin A(1) and amiloride markedly inhibited the effects of LPS.
Conclusion: The activation of inflammatory cells on metals enhanced the elution of Ni probably via the release of protons at the interface of the cells and material. (C) 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved., ELSEVIER IRELAND LTD
J. Dermatol. Sci., 2011年04月, ［査読有り］ - Suppression of intracellular calcium levels and inhibition of degranulation in RBL-2H3 mast cells by the sesquiterpene lactone parthenolide.
Hong J; Aoyama S; Hirasawa N; Zee O; Ishihara K; Hashida C; Kimura M; Seyama T; Ohuchi K, Pretreatment with parthenolide for 60 min inhibited the antigen-induced degranulation of RBL-2H3 mast cells; the IC(50) value being 4.5 +/- 0.4 mu M. The inhibition was not due to suppression of the phosphatidylinositol 3-kinase pathway because the antigen-induced phosphorylation of Akt was not inhibited by parthenolide. The antigen-induced increase in intracellular calcium levels was prevented by parthenolide, suggesting that parthenolide inhibited the antigen-induced degranulation by suppressing an increase in intracellular calcium levels. In support of this, parthenolide was found to prevent ionomycin-induced degranulation by inhibiting an increase in intracellular calcium levels. Therefore, parthenolide inhibits the degranulation of mast cells by preventing an increase in intracellular calcium levels., GEORG THIEME VERLAG KG
Planta Med., 2011年02月, ［査読有り］ - Assessment of the release of nickel from biomaterials in vivo and in vitro: enahancement by lipopolysaccharide.
Tanaka; R; Goi; Y; Ishihara; K; Ueda; K; Narushima; T; Ohtsu; H; Ohuchi; K; Hiratsuka; M; Hirasawa; N, Biodevices are implanted for long periods of time, so the release of metal ions from alloys should be tested in tissues to assess the risk of inducing metal allergies. However there is little evidence that the release of metal ions from alloys in vivo is similar to that in vitro. We implanted metal wires in mice and determined the concentration of metal ions in tissue to analyze the mechanisms responsible for metal allergies. The release of ions from the Ni wire was detected within 8 h and attained a plateau 72 h after the implantation. Furthermore, it was significantly increased by an injection of LPS. The results indicated that the release of Ni was apparently enhanced by inflammatory responses. We also established an in vitro assay system using the murine macrophage cell line RAW 264. The addition of LPS apparently increased the amount of Ni released into the medium, indicating the activation of the cells to have enhanced the elution of ions from the Ni plate. Our in vitro model using LPS-stimulated RAW264 cells might reflect the elution of Ni in inflamed tissue., The Japanese Society of Inflammation and Regeneration
Inflam. Regener., 2011年, ［査読有り］ - Salicylate restores transport function and anion exchanger activity of missense pendrin mutations.
Ishihara K; Okuyama S; Kumano S; Iida K; Hamana H; Murakoshi M; Kobayashi T; Usami S; Ikeda K; Haga Y; Tsumoto K; Nakamura H; Hirasawa N; Wada H, The SLC26A4 gene encodes the transmembrane protein pendrin, which is involved in the homeostasis of the ion concentration of the endolymph of the inner ear, most likely by acting as a chloride/bicarbonate transporter. Mutations in the SLC26A4 gene cause sensorineuronal hearing loss. However, the mechanisms responsible for such loss have remained unknown. Therefore, in this study, we focused on the function of ten missense pendrin mutations (p.P123S (Pendred syndrome), p.M147V (NSEVA), p.K369E (NSEVA), p.A372V (Pendred syndrome/NSEVA), p.N392Y (Pendred syndrome), p.C565Y (NSEVA), p.S657N (NSEVA), p.S666F (NSEVA), p.T721M (NSEVA) and p.H723R (Pendred syndrome/NSEVA)) reported in Japanese patients, and analyzed their cellular localization and anion exchanger activity using HEK293 cells transfected with each mutant gene. Immunofluorescent staining of the cellular localization of the pendrin mutants revealed that p.K369E and p.C565Y, as well as wild-type pendrin, were transported to the plasma membrane, while 8 other mutants were retained in the cytoplasm. Furthermore, we analyzed whether salicylate, as a pharmacological chaperone, restores normal plasma membrane localization of 8 pendrin mutants retained in the cytoplasm to the plasma membrane. Incubation with 10 mM of salicylate of the cells transfected with the mutants induced the transport of 4 pendrin mutants (p.P123S, p.M147V, p.S657Y and p.H723R) from the cytoplasm to the plasma membrane and restored the anion exchanger activity. These findings suggest that salicylate might contribute to development of a new method of medical treatment for sensorineuronal hearing loss caused by the mutation of the deafness-related proteins, including pendrin. (C) 2010 Elsevier B.V. All rights reserved., ELSEVIER SCIENCE BV
Hear. Res., 2010年12月01日, ［査読有り］ - Involvement of prostaglandins and histamine in nickel wire-induced acute inflammation in mice.
Hirasawa N; Goi Y; Tanaka R; Ishihara K; Ohtsu H; Ohuchi K, The irritancy of Nickel (Ni) ions has been well documented clinically. However, the chemical mediators involved in the acute inflammation induced by solid Ni are not fully understood. We used the Ni wire-implantation model in mice and examined roles of prostaglandins and histamine in plasma leakage in the acute phase. The subcutaneous implantation of a Ni wire into the back of mice induced plasma leakage from 8 to 24 h and tissue necrosis around the wire at 3 days, whereas the implantation of an aluminum wire induced no such inflammatory responses. An increase in the mRNA for cyclooxygenase (COX)-2 and HDC in cells around the Ni wire was detected 4 h after the implantation. The leakage of plasma at 8 h was inhibited by indomethacin in a dose-dependent manner. Dexamethasone and the p38 MAP kinase inhibitor SB203580 also inhibited the exudation of plasma consistent with the inhibition of the expression of COX-2 mRNA. Furthermore, plasma leakage was partially but siginificantly reduced in histamine H1 receptor knockout mice and histidine decarboxylase (HDC) knockout mice but not in H2 receptor knockout mice. These results suggested that the Ni ions released from the wire induced the expression of COX-2 and HDC, resulting in an increase in vascular permeability during the acute phase of inflammation. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res 93A: 1306-1311,2010, WILEY-LISS
J Biomed Mater Res A., 2010年06月15日, ［査読有り］ - Salicylate-induced translocation of prestin having mutation in the GTSRH sequence to the plasma membrane.
Kumano S; Iida K; Ishihara K; Murakoshi M; Tsumoto K; Ikeda K; Kumagai I; Kobayashi T; Wada H, Prestin is a key molecule for mammalian hearing. The present study investigated changes in characteristics of prestin by culturing prestin-transfected cells with salicylate, an antagonist of prestin. As a result, the plasma membrane localization of prestin bearing a mutation in the GTSRH sequence, which normally accumulates in the cytoplasm, was recovered. Moreover, the nonlinear capacitance of the majority of the mutants, which is a signature of prestin activity, was also recovered. Thus, the present study discovered a new effect of salicylate on prestin, namely, the promotion of the plasma membrane expression of prestin mutants in an active state.
FEBS Lett., 2010年04月11日, ［査読有り］ - Enhancement of ligand-dependent down-regulation of glucocorticoid receptor by lipopolysaccharide.
Hirasawa N; Yashima K; Ishihara K
Life Sci., 2009年09月02日, ［査読有り］ - Analysis of the mechanism for the development of allergic skin inflammation and the application for its treatment:establishment of a modified allergic dermatitis model in mouse ear lobes by application of 12-O-tetradecanoyl phorbol 13-acetate: putative in
Hirasawa N; Ohsawa Y; Ishihara K; Seyama T; Hong J; Ohuchi K, We established a novel allergic dermatitis model in mouse ear lobes in which antigen-nonspecific inflammation was induced by painting 12-O-tetradecanoylphorbol 13-acetate (TPA) between sensitization and challenge with picryl chloride (PiCl). This model has an advantage for analyzing atopic dermatitis-like inflammation within a short period. Analysis of the time course changes in the PiCl-induced swelling showed that the allergic inflammation was shifted from a delayed-type response to a biphasic response consisting of a weak immediate-phase response and a late-phase response by painting with TPA. The application of TPA increased the PiCl-induced infiltration of eosinophils and mast cells at the inflammatory site and shifted the cytokine milieu from Th1 to Th2. The expression of the Th2-inducing cytokine thymic stromal lymphopoietin (TSLP) mRNA was also increased by TPA. These findings suggested that the induction of antigen-nonspecific inflammation by TPA before the antigen challenge enhanced the Th2 response and modified the PiCl-induced delayed type-hypersensitivity. Using this model, we clarified that histamine plays significant roles in the early-phase swelling via H(1) receptors and the late-phase swelling via H(3)/H(4) receptors. Thus, we suggested the usefulness of the combined treatment with an H(1) antagonist and an H(4) antagonist for the suppression of atopic dermatitis., JAPANESE PHARMACOLOGICAL SOC
J Pharmacol Sci., 2009年07月, ［査読有り］ - Suppression of the antigen-stimulated RBL-2H3 mast cell activation by Artekeiskeanol A.
Hong J; Sasaki H; Hirasawa N; Ishihara K; Kwak JH; Zee O; Schmitz FJ; Seyama T; Ohuchi K, Effects of artekeiskeanol A, a newly isolated coumarin derivative from Artemisa keiskeana Miq. (Compositae), the extract of which is used for treatment of rheumatoid arthritis as a folk medicine, on the antigen-induced activation of RBL-2H3 cells were examined. RBL-2H3 cells were sensitized with dinitrophenol (DNP)-specific IgE, and then stimulated with the antigen DNP-conjugated human serum albumin (DNPHSA). Artekeiskeanol A at 10 to 100μM inhibited the antigen-induced degranulation in a concentration-dependent manner, the IC 50 value being 38.0±0.2μM. Degranulation induced by thapsigargin or A23187 also was inhibited by artekeiskeanol A at 10 to 100μM. The antigen-induced increase in the levels of mRNA for tumor necrosis factor (TNF)-α and interleukin (IL)-13 and phosphorylations of Akt, p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK) and p44/42 MAPK were also suppressed by artekeiskeanol A. Our findings suggested that the effectiveness of the extract of A.keiskeana might partly be due to the inhibition of mast cell activation by artekeiskeanol A. © Georg Thieme Verlag KG Stuttgart.
Planta Med., 2009年07月01日, ［査読有り］ - Effects of nickel on eosinophil survival.
Ishihara K; Goi Y; Hong JJ; Seyama T; Ohtsu H; Wada H; Ohuchi K; Hirasawa N, Background: Accessories, watches, coins and other items containing metal sometimes cause contact dermatitis and metal allergy. Among metals, nickel in alloys is ionized by sweat on the surface of the skin and exhibits particularly marked irritancy and allergenicity. Although eosinophils play important roles in allergy, the effects of nickel on eosinophils have not been elucidated. Methods: Eosinophils were prepared from the peritoneal cavity in rats immunized with Ascaris suum extract. Purified rat eosinophils were incubated in the presence of various kinds of metals including nickel. The viability of eosinophils was analyzed using a flow cytometer. Results: When rat eosinophils were incubated for 3 days in the presence of nickel chloride at 30-1,000 mu M, the viability of eosinophils was decreased in a concentration-dependent manner. Nickel chloride at 300 mu M significantly increased the percentage of annexin V(+) PI(-) eosinophils. The population of annexin V(+) PI(-) eosinophils was also increased by nickel sulfate, cobalt chloride and zinc sulfate. The binding of nickel ions to eosinophils was detected by flow cytometer. Conclusions: Nickel ions bind to eosinophils and decrease the viability of eosinophils through the induction of apoptosis. Nickel ions may exhibit activity which modifies the function of eosinophils in allergy. Copyright (c) 2009 S. Karger AG, Basel, KARGER
Int. Arch. Allergy Immunol., 2009年06月03日, ［査読有り］ - Modification of picryl chloride-induced allergic dermatitis model in mouse ear lobes by 12-o-tetradecanoylphorbol 13-acetate, and analysis for roles of histamine in the modified model.
Noriyasu Hirasawa; Yuhsuke Ohsawa; Goh Katoh; Kazue Shibata; Kenji Ishihara; Toshio Seyama; Souichiro Tamura; JangJa Hong; Kazuo Ohuchi, Background: In atopic dermatitis, inflammation induced by antigen-nonspecific stimuli further enhances the allergic inflammation. However, there is no experimental model in which allergic dermatitis is evoked where the inflammation has been induced by antigen-nonspecific stimuli. Here, we established a novel dermatitis model in mice and analyzed the role of histamine. Methods: After sensitization with picryl chloride (PiCl) by painting on ear lobes of cyclophosphamide-treated mice, 12-O-tetradecanoylphorbol 13-acetate (TPA) was painted twice at the same site, and then allergic inflammation was induced by painting PiCl. Histamine antagonists and cyclosporine A (CsA) were administered intravenously. Results: The application of TPA shifted the PiCl-induced allergic inflammation from a delayed-type response to a biphasic response, increased the infiltration of eosinophils and mast cells at the inflammatory site, shifted the cytokine milieu from Th1 to Th2 and induced the expression of thymic stromal lymphopoietin in the ear lobes. The PiCl-induced increase in the thickness of the ear lobe in the immediate phase was suppressed by the H1 antagonist pyrilamine. In contrast, the increase in the swelling in the late phase and the infiltration of eosinophils were suppressed by the H3/H4 antagonist thioperamide. The inhibitory effect of the combined treatment with pyrilamine and thioperamide on the TPA-modified contact dermatitis was as potent as that of CsA. Conclusion: Induction of the antigen-nonspecific inflammation by TPA enhanced the PiCl-induced allergic inflammation. Histamine plays significant roles in the earlyphase swelling via H1 receptors, and the late-phase swelling via H3/H4 receptors in this TPA-modified allergic dermatitis model. Copyright (C) 2008 S. Karger AG, Basel, KARGER
Int. Arch. Allergy Immunol., 2009年, ［査読有り］ - Effects of hyperin, isoquercitrin and quercetin onlipopolysaccharide-induced nitrite production in rat peritoneal macrophages.
Sanghyun Lee; Hee-Seung Park; Yohko Notsu; Hyun Seung Ban; Yong Pil Kim; Kenji Ishihara; Noriyasu Hirasawa; Sang Hoon Jung; Yeon Sil Lee; Soon Sung Lim; Eun-Hee Park; Kuk Hyun Shin; Toshio Seyama; JangJa Hong; Kazuo Ohuchi, The extract of the root of Acanthopanax chiisanensis Nakai is used for the treatment of inflammation. To analyse the action mechanism of this extract, the effect of hyperin (quercetin-3-O-beta-D-galactose) isolated from the ethyl acetate fraction of the root of A. chiisanensis on nitrite production and induction of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS, 1 mu g/mL)-stimulated rat peritoneal macrophages were examined. The effect of the structurally related compounds, isoquercitrin (quercetin-3-O-beta-glucose) and quercetin (an aglycone of the two compounds) isolated from the extract of the leaves of Vaccinium koreanum Nakai was also examined to compare the effect. It was shown that hyperin inhibited the LPS-induced iNOS expression and nitrite production. Of the three compounds, quercetin showed the most potent inhibitory activity. The phosphorylation of p44/42 mitogen activated protein kinase (MAPK), p38 MAPK and c-Jun N-terminal kinase (JNK) were also inhibited by these compounds. These findings suggested that hyperin in the extract of the root of A. chiisanensis inhibits nitric oxide (NO) production through inhibition of the expression of iNOS by attenuation of p44/p42 MAPK, p38 MAPK and JNK, and thus participates in the antiinflammatory activity of the extract. Copyright (C) 2008 John Wiley & Sons, Ltd., JOHN WILEY & SONS LTD
Phytother. Res., 2008年, ［査読有り］ - Mechanism for the decrease in the FIP1L1-PDGFRα protein level in EoL-1 cells by histone deacetylase inhibitors.
Kenji Ishihara; Motoko Kaneko; Hajime Kitamura; Aki Takahashi; Toshio Seyama; JangJa Hong; Koji Iida; Hiroshi Wada; Noriyasu Hirasawa; Kazuo Ohuchi, Background: Acetylation and deacetylation of proteins occur in cells in response to various stimuli, and are reversibly catalyzed by histone acetyltransferase and histone deacetylase (HDAC), respectively. EoL-1 cells have an FIP1L1-PDGFRA fusion gene that causes transformation of eosinophilic precursor cells into leukemia cells. The HDAC inhibitors apicidin and n-butyrate suppress the proliferation of EoL-1 cells and induce differentiation into eosinophils by a decrease in the protein level of FIP1L1-PDGFR alpha without affecting the mRNA level for FIP1L1-PDGFRA. In this study, we analyzed the mechanism by which the protein level of FIP1L1-PDGFR alpha is decreased by apicidin and n-butyrate. Methods: EoL-1 cells were incubated in the presence of the HDAC inhibitors apicidin, trichostatin A or n-butyrate. The protein levels of FIP1L1-PDGFR alpha and phosphorylated eIF-2 alpha were determined by Western blotting. Actinomycin D and cycloheximide were used to block RNA synthesis and protein synthesis, respectively, in the chasing experiment of the amount of FIP1L1-PDGFR alpha protein. Results: When apicidin- and n-butyrate-treated EoL-1 cells were incubated in the presence of actinomycin D, the decrease in the protein level of FIP1L1-PDGFR alpha was significantly enhanced when compared with controls. In contrast, the protein levels were not changed by cycloheximide among these groups. Apicidin and n-butyrate induced the continuous phosphorylation of eIF-2 alpha for up to 8 days. Conclusions: The decrease in the level of FIP1L1-PDGFR alpha protein by continuous inhibition of HDAC may be due to the decrease in the translation rate of FIP1L1-PDGFRA. Copyright (C) 2008 S. Karger AG, Basel., KARGER
Int. Arch. Allergy Immunol., 2008年, ［査読有り］ - Mechanisms for the proliferation of eosinophilic leukemia cells by FIP1L1-PDGFRα.
Kenji Ishihara; Hajime Kitamura; Kenji Hiraizumi; Motoko Kaneko; Aki Takahashi; OkPyo Zee; Toshio Seyama; JangJa Hong; Kazuo Ohuchi; Noriyasu Hirasawa, The constitutively activated tyrosine kinase Fip1-like 1 (FIP1L1)-platelet-derived growth factor receptor alpha (PDGFR alpha) causes eosinophilic leukemia EoL-1 cells to proliferate. Recently, we demonstrated that histone deacetylase inhibitors suppressed this proliferation and induced the differentiation of EoL-1 cells into eosinophils in parallel with a decrease in the level of FIP1L1-PDGFR alpha. In this study, we analyzed the mechanism by which FIP1L1-PDGFR alpha induces the proliferation and whether the suppression of cell proliferation triggers the differentiation into eosinophils. The FIP1L1-PDGFR alpha inhibitor imatinib inhibited the proliferation of EoL-1 cells and decreased the level of the oncoprotein c-Myc as well as the phosphorylation of extracellular signal-regulated kinase and c-Jun N-terminal kinase (JNK). The proliferation of EoL-1 cells and expression of c-Myc were also inhibited by the MEK inhibitor U0126 and JNK inhibitor SP600125. The expression of the eosinophilic differentiation marker CCR3 was not induced by imatinib. These findings suggest that FIP1L1-PDGFR alpha induces the proliferation of EoL-1 cells through the induction of c-Myc expression via ERK and JNK signaling pathways, but is not involved in the inhibition of differentiation toward mature eosinophils. (c) 2007 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
Biochem. Biophys. Res. Commun., 2008年, ［査読有り］ - Involvement of Na+/H+ exchangers in induction of cyclooxygenase-2 by vacuolar-type (H+)-ATPase inhibitors in RAW 264 cells.
Fumitaka Kamachi; Maiko Yanai; Hyun Seung Ban; Kenji Ishihara; JangJa Hong; Kazuo Ohuchi; Noriyasu Hirasawa, In the mouse macrophage-like cell line RAW 264, vacuolar-type (H+)-ATPase (V-ATPase) inhibitors, batilomycin A, and concanamycin A, increased the level of cyclooxygenase (COX)-2 protein and its mRNA. The V-ATPase inhibitor-induced expression of COX-2 was suppressed by inhibitors of c-jun N-terminal kinase (JNK) and nuclear factor-kappa B, and by inhibitors of Na+/H+ exchangers (NHEs). The batilomycin A(1)-induced activation of JNK but not degradation of IKB-alpha was suppressed by NHE inhibitors and by an inhibitor of Na+/ Ca2+ exchanger SN-6. These results suggested that V-ATPase inhibitors induce the expression of COX-2 via NHE-dependent and -independent pathways. (c) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., ELSEVIER SCIENCE BV
FEBS lett., 2007年, ［査読有り］ - Differentiation of eosinophilic leukemia EoL-1 cells into eosinophils induced by histone deacetylase inhibitors.
Kenji Ishihara; Aki Takahashi; Motoko Kaneko; Hiroki Sugeno; Noriyasu Hirasawa; JangJa Hong; OkPyo Zee; Kazuo Ohuchi, EoL-1 cells differentiate into eosinophils in the presence of n-butyrate, but the mechanism has remained to be elucidated. Because n-butyrate can inhibit histone deacetylases, we hypothesized that the inhibition of histone deacetylases induces the differentiation of EoL-1 cells into eosinophils. In this study, using n-butyrate and two other histone deacetylase inhibitors, apicidin and trichostatin A, we have analyzed the relationship between the inhibition of histone deacetylases and the differentiation into eosinophils in EoL-1 cells. It was demonstrated that apicidin and n-butyrate induced a continuous acetylation of histones H4 and 143, inhibited the proliferation of EoL-1 cells without attenuating the level of FIP1L1-PDGFRA mRNA, and induced the expression of markers for mature eosinophils such as integrin beta 7, CCR1, and CCR3 on EoL-1 cells, while trichostatin A evoked a transient acetylation of histones and induced no differentiation into eosinophils. These findings suggest that the continuous inhibition of histone deacetylases in EoL-1 cells induces the differentiation into mature eosinophils. (c) 2007 Elsevier Inc. All rights reserved., PERGAMON-ELSEVIER SCIENCE LTD
Life Sci., 2007年, ［査読有り］ - Mechanism for the differentiation of EoL-1 cells into eosinophils by histone deacetylase inhibitors.
Motoko Kaneko; Kenji Ishihara; Aki Takahashi; JangJa Hong; Noriyasu Hirasawa; OkPyo Zee; Kazuo Ohuchi, Background: EoL-1 cells have a FIP1L1-PDGFRA fusion gene which causes the transformation of eosinophilic precursor cells into leukemia cells. Recently, we suggested that the induction of differentiation of EoL-1 cells into eosinophils by the HDAC inhibitors apicidin and n-butyrate is due to the continuous inhibition of HDACs. However, neither apicidin nor n-butyrate inhibited the expression of FIP1L1-PDGFRA mRNA, although both these inhibitors suppressed cell proliferation. Therefore, in this study, we analyzed whether the levels of FIP1L1-PDGFR alpha protein and phosphorylated-Stat5 involved in the signaling for the proliferation of EoL-1 cells are attenuated by HDAC inhibitors. Methods: EoL-1 cells were incubated in the presence of apicidin, TSA or n-butyrate. FIP1L1-PDGFR alpha and phosphorylated-Stat5 were detected by Western blotting. Results: Treatment of EoL-1 cells with apicidin at 100 nM or n-butyrate at 500 M decreased the levels of FIP1L1-PDGFR alpha protein and phosphorylated-Stat5, while that with trichostatin A at 30 nM did not. Conclusions: The decrease in the level of FIP1L1-PDGFR alpha protein caused by apicidin and n-butyrate might be one of the mechanisms by which EoL-1 cells are induced to differentiate into eosinophils by these HDAC inhibitors. Copyright (C) 2007 S. Karger AG, Basel., KARGER
Int. Arch. Allergy Immunol., 2007年, ［査読有り］ - Inhibition of bone resorption in cultures of mouse calvariae by apicularen A.
JangJa Hong; Hirokazu Sasaki; Kazuaki Niikura; Maiko Yanai; Yasuhiro Nakano; Aya Yokomakura; Kenji Ishihara; Young-Sook Kang; Kazuo Ohuchi, Apicularens A and B were isolated from the myxobacterial genus Chondromyces apiculatus JW184. Apicularen A inhibited bafilomycin A1-sensitive ATP-dependent proton transport into microsome vesicles more potently than apicularen B. Bone resorption in cultures of mouse calvariae induced by human parathyroid hormone (PTH) or interleukin-1 beta (IL-1 beta) was inhibited by apicularen A at 10 and 100 nM, while apicularen B had no effect. The bisphosphonate incadronate inhibited bone resorption at 100 nM, being less effective than apicularen A. Our findings indicate that apicularen A inhibits bone resorption induced by PTH or IL-1 beta more potently than apicularen B, probably due to inhibition of the V-ATPase., GEORG THIEME VERLAG KG
Planta Med., 2007年, ［査読有り］ - Nitric oxide production by the vacuolar-type (H+)-ATPase inhibitors bafilomycin A1 and concanamycin A and its possible role in apoptosis in RAW 264.7 cells.
JangJa Hong; Yasuhiro Nakano; Aya Yokomakura; Kenji Ishihara; Soonjoo Kim; Young-Sook Kang; Kazuo Ohuchi, In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type (H+)-ATPase (V-ATPase) inhibitors bafilomycin A1 and concanamycin A induced nitric oxide (NO) production through the expression of inducible nitric-oxide synthase mRNA and its protein and decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Bafilomycin A1 and concanamycin A activated nuclear factor (NF)-kappa B and activator protein-1 and decreased the level of I kappa B-alpha and increased that of phosphorylated c-Jun N-terminal kinase (JNK). NO production induced by these V-ATPase inhibitors was suppressed by the NF-kappa B inhibitor Bay 11-7082 [ (E)3-[ (4-methylphenyl)sulfonyl])2- propenenitrile] and the JNK inhibitor SP600125 [anthra[1,9cd] pyrazol-6(2H)-one] in parallel with the partial alleviation of the V-ATPase inhibitor-induced decrease in MTT response. The Na+, K+-ATPase inhibitor dibucaine and the F-ATPase inhibitor oligomycin did not induce NO production at which concentrations the MTT response was decreased. The NO donor S-nitrosoN- acetyl-DL-penicillamine further lowered the V-ATPase inhibitor- induced decrease in the MTT response, and the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline1- oxyl-3-oxide, sodium salt (carboxy-PTIO) alleviated it partially. Mitochondrial depolarization, an index of apoptosis, was induced by bafilomycin A1 and concanamycin A. On treatment with the nitric-oxide synthase inhibitor N-G-monomethyl-L- arginine acetate, the disruption of mitochondrial membrane potential induced by bafilomycin A1 and concanamycin A was alleviated partially in parallel with the decrease in NO production. Carboxy-PTIO also alleviated it partially. Our findings suggest that the V-ATPase inhibitors bafilomycin A1 and concanamycin A similarly induce NO production and the newly produced NO participates partially in the V-ATPase inhibitor-induced apoptosis in RAW 264.7 cells., AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS
J. Pharmacol. Exp. Ther., 2006年, ［査読有り］ - Histone deacetylase inhibitors induce the differentiation of eosinophilic leukemia EoL-1 cells into eosinophhils.
Kenji Ishihara; JangJa Hong; Motoko Kaneko; Aki Takahashi; Hiroki Sugeno; Young-Sook Kang; Kazuo Ohuchi
J. Appl. Pharmacol., 2006年, ［査読有り］ - Inhibition of vacuolar-type (H+)-ATPase by the cytostatic macrolide apicularen A and its role in apicularen A-induced apoptosis in RAW 264.7 cells.
JangJa Hong; Aya Yokomakura; Yasuhiro Nakano; Kenji Ishihara; Makoto Kaneda; Mitsue Onodera; Ken-ichi Nakahama; Ikuo Morita; Jong-Woong Ahn; OkPyo Zee; Kazuo Ohuchi, Apicularen A and the known vacuolar-type (H+)-ATPase (V-ATPase) inhibitor bafilomycin A(1) induced apoptosis of RAW 264.7 cells, while apicularen B, an N-acetyl-glucosamine glycoside of apicularen A, was far less effective. Apicularen A inhibited vital staining with acridine orange of the intracellular organelles of RAW 264.7 cells, inhibited the ATP-dependent proton transport into inside-out microsome vesicles, and inhibited the bafilomycin A(1)-sensitive ATP hydrolysis. The IC50 values of the proton transport were 0.58 nM for apicularen A, 13 nM for apicularen B, and 0.95 nM for bafilomycin A(1). Furthermore, apicularen A inhibited the bafilomycin A(1)-sensitive ATP hydrolysis more potently than apicularen B. F-ATPase and P-ATPase were not inhibited by apicularen A. We concluded that apicularen A inhibits V-ATPase, and thus induces apoptosis in RAW 264.7 cells. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., WILEY
FEBS Lett., 2006年, ［査読有り］ - [The role of eosinophils in allergic inflammation].
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi
Nihon yakurigaku zasshi. Folia pharmacologica Japonica, 2005年05月, ［査読有り］, ［招待有り］ - Induction of nitric oxide production by the cytostatic macrolide apicularen A [2,4-heptadienamide, N-[ (1E)-3-[ (3S,5R,7R,9S)-3,4,5,6,7,8,9,10-octahydro-7,14 dihydroxy-1-oxo-5,9-epoxy-1H-2-benzoxacyclododecin-3-yl]-1 propenyl]-, (2Z,4Z)-(9CI)] and possible role of nitric oxide in apicularen A-induced apoptosis in RAW 264.7 cells.
JangJa Hong; Aya Yokomakura; Yasuhiro Nakano; Hyun Seung Ban; Kenji Ishihara; Jong-Woong Ahn; OkPyo Zee; Kazuo Ohuchi, We previously reported that apicularen A [2,4-heptadienamide, N-[ (1E)-3-[ (3S,5R,7R,9S)-3,4,5,6,7,8,9,10-octahydro-7,14 dihydroxy-1-oxo-5,9-epoxy-1H-2-benzoxacyclododecin-3-yl]-1 propenyl]-, (2Z,4Z)-(9CI)], a highly cytostatic macrolide isolated from the myxobacterial genus Chondromyces, induces apoptosis in the mouse leukemic monocyte cell line RAW 264.7. To analyze the action mechanism of apicularen A for the induction of apoptosis, effects of apicularen A on nitric oxide (NO) production in RAW 264.7 cells were examined. It was demonstrated that apicularen A at 10 and 100 nM induced nitrite production, whereas apicularen B [2,4-heptadienamide, N-[ (1E)-3-[ (3S,5R,7R,9S)-7-[[2-(acetylamino)-2-deoxy-beta-d-glucopyranosyl]oxy]-3,4,5,6,7,8,9,10-octahydro-14-hydroxy-1-oxo-5,9-epoxy-1H-2-benzoxacyclododecin-3-yl]-1 propenyl]-, (2Z,4Z)-(9CI)], an N-acetyl-glucosamine glycoside of apicularen A, had no effect at 100 nM. The apicularen A-induced nitrite production was accompanied by an increase in the level of inducible nitric-oxide synthase (iNOS) and its mRNA and was suppressed by the NOS inhibitor N(G)-monomethyl-l-arginine acetate (l-NMMA). In addition, apicularen A activated nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1) and decreased the level of IkappaB-alpha and increased that of phosphorylated c-Jun N-terminal kinase (JNK). Furthermore, the apicularen A-induced nitrite production was suppressed by the NF-kappaB inhibitor Bay 11-7082 [ (E)-3-(4-methylphenylsulfonyl)-2-propenenitrile] and the JNK inhibitor SP600125 [anthra[1,9-cd]pyrazol-6(2H)-one]. These findings suggested that apicularen A activates NF-kappaB and AP-1, thus triggering the expression of iNOS mRNA and iNOS protein and induces NO production. Finally, apicularen A decreased cell growth and survival and cell viability and disrupted the mitochondrial membrane potential. The addition of l-NMMA partially recovered the apicularen A-induced decrease in cell growth and survival and cell viability and the disruption of mitochondrial membrane potential. These findings suggested that NO produced by apicularen A treatment participate partially in the apicularen A-induced apoptosis in RAW 264.7 cells.
The Journal of pharmacology and experimental therapeutics, 2005年03月, ［査読有り］ - Induction of nitric oxide production by the cytostatic macrolide apicularen A [2,4-heptadienamide, N-[ (1E)-3-[ (3S,5R,7R,9S)-3,4,5,6,7,8,9,10-octahydro-7, 14 dihydroxy-1-oxo-5,9-epoxy-1H-2-benzoxacyclododecin-3-yl]-1 propenyl]-, (2Z,4Z)-(9CI)] and possible role of nitric oxide in apicularen A-induced apoptosis in RAW 264.7 cells.
JangJa Hong; Aya Yokomakura; Yasuhiro Nakano; Hyun Seung Ban; Kenji Ishihara; Jong Woong Ahn; OkPyo Zee; Kazuo Ohuchi
J. Pharmacol. Exp. Ther., 2005年, ［査読有り］ - Induction of apoptosis by apicularen A in human promyelocytic leukemia cell line HL-60
JangJa Hong; Kenji Ishihara; OkPyo Zee; Kazuo Ohuchi, Apicularen A, a macrolide isolated from the myxobacterial genus Chondromyces, suppressed the proliferation of human promyelocytic leukemia cells (HL-60 cells), increased the release of lactate dehydrogenase and induced condensation and fragmentation of chromatin at 1 to 100 nM. In addition, it induced the DNA fragmentation, increased the percentage of annexin V-stained cells, and cleaved poly(ADP-ribose) polymerase (PARP), a substrate of caspase. In contrast, apicularen B, an N-acetylglucosamine glycoside of apicularen A, had no such effects at 100 nM. These findings indicated that apicularen A induces apoptosis in HL-60 cells by activating caspases. Phosphorylation of p44/42 MAPK, p38 MAPK and Akt was not induced by apicularen A at 100 nM, suggesting that the apicularen A-induced apoptosis in HL-60 cells is not regulated by the activation of p44/42 MAPK, p38 MAPK or Akt. Furthermore, by acridine orange staining of the cells, it was suggested that apicularen A but not apicularen B inhibits vacuolar-type H+-ATPase., GEORG THIEME VERLAG KG
Planta Med., 2005年, ［査読有り］ - Inhibition of NO production by V-ATPase inhibitors and possible participation of NO in V-ATPase inhibitor-induced decrease in growth and survival of RAW 264.7 cells.
JangJa Hong; Yasuhiro Nakano; Aya Yokomakura; Kenji Ishihara; OkPyo Zee; Kazuo Ohuchi, BIRKHAUSER VERLAG AG
Inflamm. Res., 2005年, ［査読有り］ - Possible mechanism of action of the histone deacetylase inhibitors for the induction of differentiation of HL-60 clone 15 cells into eosinophils.
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi, I We have examined the effect of the histone deacetylase inhibitors apicidin, trichostatin A (TSA) and n-butyrate on the histone acetylation and the differentiation of human eosinophilic leukemia HL-60 clone 15 cells into eosinophils.
2 Viability of the cells incubated with apicidin (100 nm), TSA (30 nm) or n-butyrate (500 muM) did not change significantly, but higher concentrations of apicidin (greater than or equal to 300 nM) or TSA (greater than or equal to 100 nM) decreased the viability when examined at day 1.
3 Apicidin (100 nM) as well as n-butyrate (500 muM) induced continuous acetylations of historic H4 and lysine(14) residue on histone H3, while TSA (30 nm) induced transient acetylations.
4 After 6 days incubation, eosinophilic cells stained by Luxol-fast-blue were generated by apicidin (100 nm) and n-butyrate (500 pm) but not by TSA (30 nm). Other markers for differentiation into eosinophils such as changes in intracellular structure, and expressions of integrin beta7 and major basic protein, and the inhibition of cell proliferation were also induced by apicidin and n-butyrate but not by TSA.
5 Continuous acetylation of historic H4 achieved by repeated treatment with TSA (30 nm) at an interval of 12 h for more than three times induced such changes when examined on day 6. In addition, the induction was impaired by shortening the period of incubation with apicidin (100 nm) or n-butyrate (500 muM).
6 CCAAT/enhancer binding protein was continuously activated by apicidin (100 nM) and n-butyrate (500 muM), but was transiently activated by TSA (30 nM).
7 These findings suggest that the continuous acetylation of histories H3 and H4 is necessary for the differentiation of HL-60 clone 15 cells into eosinophils., NATURE PUBLISHING GROUP
Br. J. Pharmacol., 2004年, ［査読有り］ - Induction of apoptosis of RAW 264.7 cells by the cytostatic macrolide apicularen A.
JangJa Hong; Kouya Yamaki; Kenji Ishihara; Jong Woong Ahn; OkPyo Zee; Kazuo Ohuchi, In RAW 264.7 cells, a mouse leukaemic monocyte cell line, apicularen A decreased cell growth and survival as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner at 10-1000 nm. Apicularen B, an N-acetyl-glucosamine glycoside of apicularen A, was 10-100-fold less effective than apicularen A. Apicularen A induced a DNA ladder, an increase in the percentage of sub-G, cells and annexin V-binding cells, and promoted the activation of caspase as revealed by the cleavage of poly(ADP-ribose) polymerase, indicating that apicularen A induced apoptosis in RAW 264.7 cells. In addition, apicularen A phosphorylated p44/42 mitogen-activated protein kinase (MAPK) and p38 MAPK. The p44/42 MAPK inhibitor PD98059 rescued the cells from apicularen-induced decrease in cell growth and survival as determined by the MTT assay, while the p38 MAPK inhibitor SB203580 augmented the effect of apicularen A. This suggested the activation of p44/42 MAPK to be pro-apoptotic and the activation of p38 MAPK antiapoptotic in apicularen A-treated RAW 264.7 cells., ROYAL PHARMACEUTICAL SOC GREAT BRITAIN
J. Pharm. Pharmacol., 2003年, ［査読有り］ - Preparation of recombinant rat eosinophil-associated ribonuclease-1 and -2 and analysis of their biological activitites.
Kenji Ishihara; Kanako Asai; Masahiro Nakajima; Suetsugu Mue; Kazuo Ohuchi, Rat eosinophils contain eosinophil-associated ribonucleases (Ears) in their granules. Ears are thought to be synthesized as pre-forms and stored in the granules as mature forms. However, the N-terminal amino acid of mature Ear-1 and Ear-2 is still controversial. Therefore, we prepared two recombinant mature forms of Ear-l and Ear-2 in which the N-terminal amino acids are Ser(24) (S) [Ear-1 (S) and Ear-2 (S)] and Gln(26), (Q) [Ear-1 (Q) and Ear-2 (Q)], and analyzed their biological activities by comparing them with those of pre-form Ear-1 and pre-forrn Ear-2. The four mature Ears showed RNase A activity as well as bovine pancreatic RNase A activity, but pre-Ear-1 and pre-Ear-2 showed no RNase A activity. Mature Ear-1 (Q) and mature Ear-2 (Q) showed more potent RNase A activity than mature Ear-1 (S) and mature Ear-2 (S), respectively. The RNase A activities of mature Ear-1 (Q) and mature Ear-2 (Q) were reduced by treatment at 96degreesC for 20 min or with RNase inhibitor. The growth of Escherichia coli was inhibited by both pre-Ears and mature Ears in a concentration-dependent manner, and was almost completely suppressed at 1.0 muM, The bactericidal activities of mature Ear-1 (Q) and mature Ear-2 (Q) were not inhibited by RNase inhibitor, but was increased by treatment at 96degreesC for 20 min. (C) 2003 Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
Biochim. Biophys. Acta, 2003年, ［査読有り］ - Apicidin, a histone deacetylase inhibitor, induces differentiation of HL-60 cells.
JangJa Hong; Kenji Ishihara; Kouya Yamaki; Kenji Hiraizumi; Tadao Ohno; Jong Woong Ahn; OkPyo Zee; Kazuo Ohuchi, The fungal metabolite apicidin (cyclo(N-O-methyl-L-tryptophanyl-L-isoleucinyl-D-pipecolinyl-L-2-amino-8-oxodecanoyl)) inhibited the growth of HL-60 cells in a concentration-dependent manner (100-1000 nM). At higher concentrations (>300 nM), cell death was induced. At 100 nM, it induced hyperacetylation of histone H4 time-dependently, while trichostatin A induced transient hyperacetylation. Apicidin (10-100 nM) increased the cells having nitroblue tetrazolium-reducing activity and expressing CD11b but not CD14 and CD15. The expression of CD11b by apicidin was long lasting, while that by trichostatin A was transient. In K562 cells, apicidin at 10-100 nM did not inhibit cell growth nor express CD11b, CD14 and CD15. Our findings indicate that apicidin inhibits proliferation and induces the early stage of differentiation of HL-60 cells. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved., ELSEVIER SCI IRELAND LTD
Cancer Lett., 2003年, ［査読有り］ - Differentiation of HL-60 clone 15 cells to eosinophils by histone deacetylase inhibitors.
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi
Inflamm. Res., 2003年, ［査読有り］ - Expression and purification of recombinant rat eosinophil-associated ribonucleases, homologues of human eosinophil cationic protein and eosinophil-derived neurotoxin, and their characterization.
Masahiro Nakajima; Mikito Hirakata; Takeaki Nittoh; Kenji Ishihara; Kazuo Ohuchi, Background. Human eosinophils contain two eosinophil ribonucleases, eosinophil cationic protein (ECP) and eosinophil-derived neurotoxin (EDN). In rats, 8 homologues of human ECP and EDN have been identified. To clarify the biological activity of rat eosinophil ribonucleases, we cloned rat eosinophil-associated ribonuclease (EAR)-1/rat ribonuclease 7 and rat EAR-2/rat ribonuclease 4, and produced recombinant rat pre-EAR-1 and pre-EAR-2 in a bacterial expression system. Methods:As we have already cloned the complete nucleotide sequence for rat EAR-1, we determined that for rat EAR-2 cDNA by the rapid amplification of cDNA ends procedure. Recombinant rat pre-EAR-1 and pre-EAR-2 were expressed in Escherichia coli as N-terminal 6 x histidine tagged proteins, isolated from the insoluble fraction of the cell lysate and purified by a single-step method using an Ni-NTA resin column after solubilization with a 6 M guanidine solution. Results: The deduced amino acid sequence revealed that the molecular weight of EAR-2 containing the signal peptide is 17.3 kD and the isoelectric point is 8.59. The homology in amino acid sequence between rat pre-EAR-2, and human pre-ECP and human pre-EDN is 51 and 53%, respectively. The purified and refolded recombinant rat pre-EAR-1 and pre-EAR-2 showed bactericidal activity against E. coli and Staphylococcus aureus. Conclusions: These findings suggest that rat EAR-1 and EAR-2 act as host defense factors against bacterial infection in rats. Copyright (C) 2001 S. Karger AG, Basel., KARGER
Int. Arch. Allergy Immunol., 2001年, ［査読有り］ - Possible participation of a JAK2 signaling pathway in recombinant rat interleukin-5-induced prolongation of rat eosinophil survival.
Kenji Ishihara; Ikuko Satoh; Suetsugu Mue; Kazuo Ohuchi, Recombinant rat interleukin (IL)-5-induced prolongation of rat eosinophil survival in culture was inhibited in a concentration-dependent manner by the protein synthesis inhibitor cycloheximide, the DNA-dependent RNA synthesis inhibitor actinomycin D, and the tyrosine kinase inhibitor herbimycin A when examined 96 h after incubation. The MEK-1 inhibitor PD98059 inhibited IL-S-induced phosphorylation of both p44 and p42 MAP kinases, but the IL-S-induced prolongation of eosinophil survival was not inhibited. In contrast, the JAK2 inhibitor AG490 inhibited the IL-S-induced prolongation of eosinophil survival. Treatment of eosinophils with IL-S resulted in phosphorylation of STAT5 but not STAT1, and the IL-S-induced phosphorylation of STAT5 was inhibited by AC490. These findings suggest that the activation of JAK2 tyrosine kinase and protein synthesis are required for the prolongation of rat eosinophil survival induced by recombinant rat IL-5. STAT5 phosphorylation might also participate in the IL-S-induced survival of rat eosinophils. (C) 2001 Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
Biochim. Biophys. Acta, 2001年, ［査読有り］ - Analysis of the prolongation of rat eosinophil survival induced by recombinant rat interleukin-5.
Kenji Ishihara; Ikuko Satoh; Kazuo Ohuchi, Rat eosinophil survival was prolonged by recombinant rat IL-5 prepared by the baculovirus expression system. The IL-5-induced prolongation of eosinophil survival was dose-dependently inhibited by the protein synthesis inhibitor cycloheximide, the DNA-dependent RNA synthesis inhibitor actinomycin D, and the tyrosine kinase inhibitor herbimycin A. The MEK-1 inhibitor PD98059 inhibited IL-5-induced phosphorylation of both p44 and p42 MAP kinases, but the IL-5-induced prolongation of eosinophil survival was not inhibited. In contrast, the JAK2 inhibitor AG490 inhibited the IL-5-induced prolongation of eosinophil survival. Treatment of eosinophils with IL-5 resulted in phosphorylation of STAT5 but not STAT1, and the IL-5-induced phosphorylation of STAT5 was inhibited by AG490. These findings suggest that recombinant rat IL-5 activates JAK2 tyrosine kinase, which phosphorylates STAT5, and induces protein syn thesis required for the prolongation of rat eosinophil survival. Copyright (C) 2000 S. Karger AG, Basel., KARGER
Int. Arch. Allergy Immunol., 2000年, ［査読有り］ - Generation of rat eosinophils by recombinant rat interleukin-5 in vitro and in vivo.
Kenji Ishihara; Ikuko Satoh; Suetsugu Mue; Kazuo Ohuchi, The addition of recombinant rat interleukin-5 (IL-5), which was purified from the hemolymph of silkworm Bombyx mori larvae infected with IL-5-expressing recombinant virus, to cultures of rat bone marrow cells resulted in an increase in the number of Luxol-fast-blue staining eosinophils in a time- and concentration-dependent manner. After 6 days culture with 100 pM recombinant rat IL-5, more than 90% of the bone marrow cells were eosinophil. The contents of major basic protein (MBP) in the bone marrow cells determined by Western blot analysis using a polyclonal antibody to rat MBP were also increased by recombinant rat IL-5 (100 pM). Furthermore,, intravenous injections of recombinant rat IL-5 twice a day for six consecutive days increased the population of eosinophils in peripheral blood cells and in bone marrow cells. These findings indicate that rat IL-5 induces terminal differentiation and proliferation of progenitor cells to mature eosinophils in rats. (C) 2000 Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
Biochim. Biophys. Acta, 2000年, ［査読有り］ - Analysis of biological activities of recombinant rat interleukin-5.
Kenji Ishihara; Takeaki Nittoh; Ikuko Satoh; Kazuo Ohuchi, We analyzed the biological activities of recombinant rat interleukin-5 (IL-5). Purified recombinant rat IL-5 promoted the proliferation of T88-M cells in a concentration dependent manner, and its effect was inhibited by an anti-murine IL-5-neutralizing polyclonal antibody. When bone marrow cells from normal rats were incubated with recombinant rat IL-5 in medium containing methylcellulose, the colony formation by eosinophilic cells was induced. Furthermore, when rat peritoneal eosinophils were incubated with recombinant rat IL-5, the spontaneous decrease in eosinophil viability was inhibited in a time- and concentration-dependent manner. In addition, the recombinant rat IL-5-induced eosinophil survival was inhibited by an anti-murine IL-5-neutralizing polyclonal antibody. These findings suggest that rat IL-5 acts as B cell growth factor II, eosinophil differentiation factor, and eosinophil survival-enhancing factor., KARGER
Int. Arch. Allergy Immunol., 1999年, ［査読有り］ - Preparation of recombinant rat interleukin-5 by baculovirus expression system and analysis of its biological activities.
Kenji Ishihara; Ikuko Satoh; Takeaki Nittoh; Toshimichi Kanaya; Hironobu Okazaki; Takeo Suzuki; Teruyuki Koyama; Tamai Sakamoto; Tsuyoshi Ide; Kazuo Ohuchi, Rat interleukin-5 (IL-5) cDNA was subcloned from peritoneal cells collected 4 h after intraperitoneal injection of Ascaris suum antigen solution into the immunized rats. Cysteine proteinase-deleted (CPd) rat IL-5 recombinant virus was constructed by inserting rat IL-5 cDNA into CPd virus having a deletion in the cysteine proteinase gene of the silkworm Bombyx mori nuclear polyhedrosis virus. On infection with the CPd rat IL-5 recombinant virus, the silkworm B. mori larvae produced rat IL-5 as a dimeric form in hemolymph. Recombinant rat IL-5 was purified more than 95.5% by anion-exchange chromatography and hydrophobic chromatography. The purified recombinant rat IL-5 promoted the proliferation of T88-M cells in a concentration-dependent manner, and its effect was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. When bone marrow cells from normal rats were incubated with recombinant rat IL-5 in medium containing methylcellulose, the colony formation by eosinophilic cells was induced. Furthermore, when rat peritoneal eosinophils were incubated with recombinant rat IL-5, the spontaneous decrease in the eosinophil viability was inhibited in time- and concentration-dependent manners. In addition, the recombinant rat IL-5-induced eosinophil survival was inhibited by an anti-murine IL-5 neutralizing polyclonal antibody. These findings suggest that rat IL-5 acts as B-cell growth factor II (BCGF-II), eosinophil differentiation factor (EDF), and eosinophil survival-enhancing factor. (C) 1999 Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
Biochim. Biophys. Acta, 1999年, ［査読有り］ - Monoclonal antibody to human midkine reveals increased midkine expression in human brain tumors.
Shinsuke Kato; Kenji Ishihara; Takao Shinozawa; Hiroyuki Yamaguchi; Yoshiya Asano; Masaya Saito; Masako Kato; Tadashi Terada; Akira Awaya; Asao Hirano; Dennis W. Dickson; Shu-Hui Yen; Eisaku Ohama, We produced a rat IgG2a monoclonal antibody against the carboxyl terminal region of human midkine (MK), a novel growth factor. This monoclonal antibody was used in immunohistochemical studies to compare the expression of MK, proliferating cell nuclear antigen (PCNA) and p53 protein in 133 primary brain tumors and 21 carcinoma metastases to the central nervous system. Approximately half of the glioblastomas multiforme (GBMs) (19/32), medulloblastomas (8/14), primitive neuroectodermal tumors (PNETs) (5/11), breast carcinoma metastases (Br-Mts) (6/10) and lung carcinoma metastases (L-Mts) (5/11) as well as some astrocytomas (2/14) had tumor cells that expressed MK: however, oligodendrogliomas,ependymomas, schwannomas, meningiomas, and pituitary adenomas did not express MK. The values of the PCNA-labeling index were statistically higher in GBMs, medulloblastomas, PNETs, Br-Mts, and L-Mts that expressed MK than in those that did not (Wilcoxon rank-sum test, p < 0.05). There was no correlation between MK and p53 protein in all tumor types. Normal and non-neoplastic brain tissues were negative for MK, PCNA, and p53 protein. We conclude that primary and metastatic tumors of the brain express MK and that the MK expression in brain tumors may depend, in part, on the proliferating potential., AMER ASSN NEUROPATHOLOGISTS INC
J. Neuropathol. Exp. Neurol., 1999年, ［査読有り］ - Effects of glucocorticoids on apoptosis of infiltrated eosinophils and neutrophils in rats.
Takeaki Nittoh; Hiroko Fujimori; Yoshiko Kozumi; Kenji Ishihara; Suetsugu Mue; Kazuo Ohuchi, The effects of glucocorticoids on the survival of rat eosinophils and neutrophils infiltrated into the peritoneal cavity were examined. Glucocorticoids including dexamethasone, prednisolone and hydrocortisone inhibited the survival of rat peritoneal eosinophils at 10(-6) M, whereas they prolonged survival of rat peritoneal neutrophils at 10(-8) M. Sex steroids including estradiol and progesterone did not affect cell survival. Dexamethasone decreased the viability of eosinophils after 3 days of incubation and maintained the viability of neutrophils until 4 days after incubation concentration dependently. The EC50 of dexamethasone for inhibition of the survival of eosinophils was 1.5 x 10(-8) M, and that for the spontaneous death of neutrophils was 6.4 x 10(-10) M, suggesting that glucocorticoids at concentrations that inhibit eosinophil survival prolong neutrophil survival. Analysis of DNA fragmentation of cultured eosinophils and neutrophils revealed that glucocorticoids enhance eosinophil apoptosis but inhibit neutrophil apoptosis. The effects of dexamethasone on viability and DNA fragmentation were counteracted by the glucocorticoid receptor antagonist, mifepristone, concentration dependently. These findings indicate that glucocorticoids induce contradictory effects via the glucocorticoid receptor on rat eosinophils and neutrophils extravasated to an inflammatory locus such as the peritoneal cavity by modulating apoptosis. (C) 1998 Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
Eur. J. Pharmacol., 1998年, ［査読有り］
MISC
- 学校でのアレルギー疾患に関する取り組みについての調査
石原 研治; 鈴木 美香; 寺門 遼香; 福田 珠巳; 岩本 里美; 金子 和子; 石川 雅世; 瀧澤 利行
茨城大学教育学部 教育学部紀要（教育科学）, 2023年01月31日
筆頭著者 - 養護教諭が主体となり実施する教員及び職員対象の校内研修の実態について
大曽根 沙季; 廣原 紀恵; 石原 研治
茨城大学教育学部 教育学部紀要（教育科学）, 2023年01月31日
ラスト(シニア)オーサー - アレルギー疾患の理解を深めるための事例の作成と教職大学院での展開
石原 研治; 鈴木 美香; 寺門 遼香; 福田 珠巳; 瀧澤 利行
茨城大学教育学部 教育学部紀要（教育科学）, 2023年01月31日
筆頭著者 - 新型コロナウイルス感染症による学校の長期休業期間中における中学生の生活とコミュニケーションの実態
鈴木 美香; 石原 研治
茨城大学教育学部 教育学部紀要（教育科学）, 2023年01月31日
ラスト(シニア)オーサー - 養護教諭の救急対応における能力向上のための自己研修ツールの開発
高橋; 朋子; 古池; 雄治; 石原; 研治; 廣原; 紀恵
教育学部紀要（教育科学） = Bullentin of the,Colledge of Education,Ibaraki University,(Educational Sciences), 2021年01月 - 中学生の外科的応急処置に関連する実態
廣原; 紀恵; 澤田; 葵; 石原; 研治
教育学部紀要（教育科学） = Bullentin of the,Colledge of Education,Ibaraki University,(Educational Sciences), 2021年01月 - 再生医療に関する高校生の意識調査
石原 研治; 柴田 有沙; 鏑木 瞳; 瀧澤 利行
茨城大学教育学部紀要. 教育科学, 2018年01月30日 - 新聞等を通じた研究成果の適切な情報発信に向けた表現の検討
和田濵 裕之; 中田 祐希; 石原 研治
茨城大学教育学部紀要. 教育科学, 2018年01月30日 - 中・高校生の理解促進を指向した再生医療通信『懸け橋』の発行
石原研治・佐藤公美・野口 遥・山口千恵子・大沼守正; 佐藤 隆・和田濱裕之・川上雅弘・瀧澤利行
茨城大学教育実践研究, 2016年12月01日 - 高大接続研究による「再生医療教育モデル講座」の開発
石原研治・山口千恵子・大沼守正・中森未華・瀧澤利行
茨城大学教育実践研究, 2016年12月01日 - アレルギー疾患に対する養護教諭の保健指導の実態調査 -他の職種との連携及び学校生活管理指導表の活用の実態について‐
熊谷仁美; 竹下誠一郎; 宮川八平; 石原研治
茨城大学教育学部紀要（教育科学）, 2012年03月31日 - Induction of thymic stromal lymphopoietin by chemical compounds in vivo and exacerbation of allergy.
Sato N; Ishihara K; Hiratsuka M; Hirasawa N
Inflam. Regener., 2011年, ［査読有り］ - 内耳に発現するpendrinのバイオメカニクス
和田 仁; 石原 研治; 小山 眞
ナノ医工学年報 = Annual report / Tohoku University Global COE Program, Global Nano-Biomedical Engineering Education and Research Network Centre, 2010年 - 韓方生薬Artemisia keiskeana Miq.の抽出液中に含まれる抗アレルギー性物質の探索とその作用機序解析
佐々木 宏和; Hong JangJa; Hwan Kwak John; 石原 研治; 青木 淳賢; 平澤 典保; Schmitz Francis J; Zee OkPyo; 瀬山 敏雄; 大内 和雄
日本薬学会年会要旨集, 2008年03月 - 炎症性刺激によるグルココルチコイド受容体(GR)αの分解促進とキナーゼの関与について
八島 一史; 石原 研治; 青木 淳賢; 平澤 典保
日本薬学会年会要旨集, 2008年03月 - Lead compounds for anti-inflammatory drugs isolated from the plants of the traditional oriental medicine in Korea.
JangJa Hong; Kuku Hyun Shin; Soon Sung Lim; Jong Hwan Kwak; OkPyo Zee; Kenji Ishihara; Noriyasu Hirasawa; Toshio Seyama; Kazuo Ohuchi
Inflam. Allergy - Drug Targets, 2008年, ［査読有り］ - 抗原特異的炎症誘発によるアレルギー性皮膚炎の増悪化
平澤典保、大澤雄亮、石原研治、Hong JangJa、瀬山敏雄、大内和雄
炎症と免疫, 2008年 - Anti-inflammatory effects of Na+/H+ exchangers inhibitors.
Noriyasu Hirasawa; Fumitaka Kamachi; Maiko Yanai; Hyun Seung Ban; Kenji Ishihara; Toshio Seyama; JangJa Hong; Kazuo Ohuchi
Inflamm. Regener., 2008年, ［査読有り］ - HDAC 阻害薬による好酸球性白血病の好酸球への分化誘導機構
石原研治、大内和雄
炎症と免疫, 2005年, ［招待有り］ - アレルギー性炎症における好酸球の役割
石原研治、Hong JangJa、Zee OkPyo、大内和雄
日本薬理学会誌, 2005年, ［招待有り］ - Mechanism of the eosinophilic differentiation of HL-60 clone 15 cells induced by n-butyrate.
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi
Int. Arch. Allergy Immunol., 2005年, ［査読有り］ - アレルギー性炎症における好酸球の増多機構とその役割に関する研究
石原研治
薬学雑誌, 2005年, ［招待有り］
書籍等出版物
- 新版 学校保健 第９節 学校環境衛生
石原研治; 佐藤公美, 共著
2024年09月24日 - 学校保健「第９節 学校環境衛生」
石原研治, 単著
東山書房, 2019年04月03日 - 学校保健「第９節 学校環境衛生」
石原研治, 単著
東山書房, 2019年04月03日, ［査読有り］
9784827815702 - “Chapter 5 Eosinophil Cell Lines” Garry M. Walsh (ed.) Eosinophils Methods and Protocols
単著
Humana Press, 2014年07月01日
9781493910151 - 炎症の制御と修復 「好酸球と慢性炎症」
単著
北陸館, 2013年11月21日 - "Chapter 4.2 Eosinophil Cell Lines." In: JJ Lee and HF Rosenberg (Eds.), Eosinophils in Health and Disease.
Kenji Ishihara; Noriyasu Hirasawa; Kazuo Ohuchi, 共著
Elsevier, 2012年10月
9780123943859 - スタンダード薬学シリーズ 第４巻 生物系薬学 III. 生体防御 SOB 16 アレルギーについて分類し、担当細胞および反応機構を説明できる。SOB 17 炎症の一般的症状、担当細胞および反応機構について説明できる。
石原研治,大内和雄, 共著
東京化学同人, 2006年03月27日 - COX-2 阻害薬 Q&A COX-2 阻害薬の薬理作用とその機序：抗炎症作用は ?
共著
医薬ジャーナル社, 2003年12月05日 - 薬のサイエンス６ 非ステロイド性抗炎症薬 体内動態 -クスリは体の中でどうなっているのか ?
石原研治、大内和雄, 共著
フジメディカル出版, 2000年10月01日
講演・口頭発表等
- 当学会の中高生セッションの意義
石原研治; 川上雅弘
第23回日本再生医療学会総会, 2024年03月23日, ［招待有り］
20240321, 20240323 - 〔主要な業績〕特別支援学校の教員のアレルギー疾患に関する意識
日本アレルギー学会, 2023年10月21日
20231020, 20231022 - 小中学生における 視力検査の縦断的結果と姿勢改善による近視予防
鈴木美香,石原研治,瀧澤利行
日本学校保健学会第68回学術大会, 2022年11月04日, 一般社団法人日本学校保健学会
20221104, 20221106 - セルフメディケーションにおける薬局薬剤師の役割を啓発するボードゲームの開発
小野寺亮、富永敦子、佐藤健太、石原研治、一條宏、平澤典保
第32回日本医療薬学会, 2022年09月23日
20220923, 20220925 - 薬局薬剤師すごろく「ふぁるま ふぁんたじあ」の開発
小野寺亮、富永敦子、佐藤健太、石原研治、一條宏、平澤典保
医療薬学フォーラム2022, 2022年07月23日
20220723, 20220724 - 授業科目「メディカルサイエンス」を通した幅広い視野を有した再生医療人材育成の試み
兼龍盛 石原研治
第21回日本再生医療学会総会, 2022年03月19日, 日本再生医療学会
20220317, 20220319 - 新型コロナウイルス感染症による学校の長期休業期間中における生徒の生活とコミュニケーションの実態
鈴木美香 石原研治
日本学校保健学会 第 67 回学術大会, 2021年11月05日, 日本学校保健学会
20211105, 20211107 - 日本再生医療学会総会「中高生のためのセッション」の狙い
川上 雅弘,鈴木 一史,石原 研治
第20回日本再生医療学会総会, 2021年03月
20210311, 20210313 - 主体的・対話的で深い学びをめざす日本再生医療学会ジュニアセッションの取り組み
川上雅弘,石原研治
科学教育学会, 2020年08月
20200825, 20200827 - 再生医療に対する市民の意識調査アンケート
石原 研治; 礒嵜 友輔; 添田 風花; 高野 恭平; 小林 優吾; 大津 友萌佳; 緑川 礼夢; 山口 千恵子; 瀧澤 利行
第18回日本再生医療学会総会, 2019年03月22日
20190321, 20190323 - 教育現場から理解する再生医療
石原 研治
第18回日本再生医療学会総会 シンポジウム「オールジャパンで進めるヒトづくり～企業・教育機関・行政の立場から～」, 2019年03月23日, 第18回日本再生医療学会総会, ［招待有り］ - 医療従事者のiPS細胞と再生医療に対する関心・理解に関する質問紙調査
石津美阿里,廣原紀恵,石原研治,瀧澤利行
第 14 回日本再生医療学会総会, 2015年03月 - 中・高・大学生の iPS 細胞と再生医療に対する関心
石原研治,石津美阿里,一宮飛鳥,稲見真美,橋本朋美,湯澤理恵,廣原紀恵,八代嘉美,瀧澤利行
第 14 回日本再生医療学会総会, 2015年03月 - 中学生・高校生・大学生の iPS 細胞・再生医療に関する知識・関心・理解
石津美阿里,石原研治,瀧澤利行,廣原紀恵
第 61 回日本学校保健学会, 2014年11月 - アレルギー疾患の児童生徒への養護教諭の対応
熊谷仁美,竹下誠一郎,宮川八平,石原研治
第58回日本学校保健学会, 2011年11月 - 健常者と罹患者のアレルギー意識―アレルギー 疾患別にみる知識の違いについて―
熊谷仁美; 河田史宝; 石原研治
日本健康相談活動学会第７回学術集会, 2011年02月 - 健常者と罹患者のアレルギー意識
熊谷仁美,河田史宝,宮川八平,竹下誠一郎,石原研治
第57回日本学校保健学会, 2010年11月 - 難治性疾患の基礎医学的研究から教育学的研究へ
石原研治
茨城大学養護教諭同門会 平成2１年度「教職員のための研修会（第七回）」, 2009年07月29日 - Induction of thymic stromal lymphopoietin by chemical compounds in vivo and analysis of the producing cells.
Nozomi Satou; Kenji Ishihara; Noriyasu Hirasawa
9th World Congress on Inflammation, 2009年07月06日 - Release of nickel from the implanted wire in vivo and enhancement by lipopolysaccharide.
Rina Tanaka; Yoshiaki Goi; Kenji Ishihara; K Ueda; T Narushima; Hiroshi Ohtsu; Kazuo Ohuchi; Noriyasu Hirasawa
9th World Congress on Inflammation, 2009年07月06日 - 好酸球に対するニッケルの作用の解析
石原 研治、; JangJa HONG、平澤 典保、大津 浩、瀬山 敏雄、和田 仁、大内 和雄
日本薬学会 第 129 年会, 2009年03月28日 - 生体内における金属からのニッケル溶出の簡易評価法の確立と溶出機序の解析
田中 里奈、五井 嘉明、石原 研治、上田 恭介、成島 尚之、大津 浩、大内 和雄、平澤 典保
日本薬学会 第 129 年会, 2009年03月28日 - New strategy for rescue of pendrin mutants causing pendred syndrome.
Shuhei Okuyama; Kenji Ishihara; Shun Kumano; Koji Iida; Toshimitu Kobayashi; Shinichi Usami; Hiroshi Wada
8th International Symposium of Tohoku University Global COE Program ”Global Nano-Biomedical Engineering Education and Research Network Centre”, 2008年12月09日 - Effects of HDAC inhibitors on leukemia cells.
Kenji Ishihara; JangJa Hong; Kazuo Ohuchi; Hiroshi Wada
7th International Symposium of 2007 Tohoku University Global COE Program ”Global Nano-Biomedical Engineering Education and Research Network Centre”, 2008年10月17日 - 遺伝性難聴の原因となるペンドリン変異体のサリチル酸によるリフォールディング
奥山修平、石原研治、飯田 浩司、熊野峻、小林俊光、宇佐美真一、和田仁
第19回バイオフロンティア講演会, 2008年09月24日 - 好酸球に対するニッケルの作用
石原研治、五井嘉明、飯田浩司、瀬山利雄、Hong JangJa、大内和雄、大津浩、平澤典保、和田仁
第22 回アレルギー・好酸球研究会 2008, 2008年06月22日 - 炎症性刺激によるグルココルチコイド受容体 (GR)αの分解促進とキナーゼの関与について
八島一史、石原研治、青木淳賢、平澤典保
日本薬学会第128年会, 2008年03月26日 - 韓方生薬 Artemisia keiskeana Miq. の抽出液中に含まれる抗アレルギー性物質の探索とその作用機序解析
佐々木宏和、Hong JangJa、Kwak John Hwan、石原研治、青木淳賢、平澤典保、Schmitz Francis J、Zee OkPyo、瀬山敏雄、大内和雄
日本薬学会第128年会, 2008年03月26日 - アレルギー性疾患における好酸球増多機構とその治療戦略
石原研治、JangJa Hong、OkPyo Zee、大内和雄
日本薬学会第126年会, 2006年03月28日, ［招待有り］ - Induction of NO production by V-ATPase inhibitors and possible participation of NO in V-ATPase inhibitor-induced decrease in growth and survival of RAW 264.7 cells.
Kazuo Ohuchi; JangJa Hong; Aya Yokomakura; Yasuhiro Nakano; Kenji Ishihara; OkPyo Zee
Deutsche Pharmazeutische Gesellschaft-Jahrestagung 2005, 2005年10月05日 - Induction of NO production by V-ATPase inhibitors and possible participation of NO in V-ATPase inhibitor-induced decrease in growth and survival of RAW 264.7 cells.
JangJa Hong; Yasuhiro Nakano; Aya Yokomakura; Kenji Ishihara; OkPyo Zee; Kazuo Ohuchi
7th World Congress on Inflammation, 2005年08月19日 - Possible mechanism of action of the histone deacetylase inhibitors for the induction of differentiation of HL-60 clone 15 cells into eosinophils.
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi
4th Biennial Congress of the International Eosinophil Society, 2005年05月25日 - アレルギー性炎症における好酸球の役割 -好酸球顆粒蛋白質の役割に関する考察-
石原研治、Hong JangJa、Zee OkPyo、大内和雄
第77回日本薬理学会年会, 2004年03月10日, ［招待有り］ - Differentiation of HL-60 clone 15 cells to eosinophils by histone deacetylase inhibitors.
Kenji Ishihara; JangJa Hong; OkPyo Zee; Kazuo Ohuchi
6th World Congress on Inflammation, 2003年08月02日 - Interleukin-5 による好酸球増多誘導のメカニズム
石原研治、大内和雄
日本薬学会第120年会, 2000年03月29日, ［招待有り］ - 好酸球増多因子としての IL-5 -Recombinant ラット IL-5 を用いた解析-
石原研治、大内和雄
第 3 回免疫薬理研究会, 1999年12月18日, ［招待有り］
所属学協会
共同研究・競争的資金等の研究課題
メディア報道
学術貢献活動
- 第23回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2024年03月21日 - 2024年03月23日 - 第22回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2023年03月22日 - 2023年03月25日 - 第21回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2022年03月17日 - 2022年03月19日 - 第20回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2021年03月11日 - 2021年03月13日 - 〔主要な業績〕第19回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2020年05月01日 - 2020年05月29日 - 第18回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2019年03月21日 - 第17回日本再生医療学会「中高生のためのセッション」
企画立案・運営等
2018年03月22日
