The functional analysis of antimicrobial peptide SLPI in the intestine and a colon protective effect of Daikenchuto

• Project/Area Number: 20K16028
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47050:Environmental and natural pharmaceutical resources-related
• Research Institution: Oita University
• Principal Investigator: Ozaka Sotaro 大分大学, 医学部, 助教 (60835700)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: SLPI / 大建中湯 / 漢方薬 / DSS腸炎 / 炎症性腸疾患 / 腸炎

Outline of Research at the Start

SLPIは抗菌ペプチドの一つであり、肺や皮膚において組織保護作用が証明されているが、消化管における機能は不明である。我々はSLPIが消化管において保護作用を示すかどうかをSLPI欠損マウスを用いて証明する。具体的にはSLPI欠損マウスに腸炎モデルであるDSS腸炎を誘導し、野生型マウスとの感受性の差異を評価する。SLPI欠損マウスで腸炎が増悪した場合、SLPIが腸炎保護作用を有していることが証明できる。
また我々は大建中湯をマウスに投与すると腸管でSLPIが増加することを見出した。そこで、大建中湯にSLPI発現増強を介した腸炎改善効果があるかどうかを腸炎モデルマウスに大建中湯を投与し証明する。

Outline of Final Research Achievements

In the present study, the physiological function of secretory leukocyte protease inhibitor (SLPI) in the gut and the colon protective effect of Daikenchuto (DKT) through the induction of SLPI were demonstrated. SLPI was induced by inflammatory stimuli in the gut and protected the intestinal epithelial barrier by antiprotease activity. In addition, we found that the expression of SLPI in the colon was significantly increased in DKT-treated mice. Next, to investigate a therapeutic effect of DKT through the induction of SLPI, DKT was administered to DSS-induced colitis model. The results showed that DSS-induced colitis was alleviated by the administration of DKT in wild-type mice, but not in SLPI-deficient mice.
These results indicate that DKT exerts colon protective effect through the enhancement of SLPI, suggesting that DKT may be used as a therapeutic agent for inflammatory bowel disease.

Academic Significance and Societal Importance of the Research Achievements

本研究により抗菌ペプチドSLPIの腸管保護作用が証明され、大建中湯の抗炎症作用の一つとして、SLPIの発現増強を介した腸管保護作用があることが示唆された。近年、炎症性腸疾患(IBD)の治療薬として様々な抗体製剤が開発されているが、これら薬剤は高価で、免疫抑制による副作用の懸念がある。大建中湯は本邦で古典的に用いられてきた漢方薬で、安価なことに加え、重篤な副作用は生じない。そのため、大建中湯がIBDの治療薬として応用されれば、安価で安全性の高い薬剤が治療選択肢に加わることになり医療コストの削減も期待できる。本研究は大建中湯のIBD領域における臨床応用に向けての基礎になる研究と考えている。

Research Products

• [Journal Article] Acute Cholecystitis Associated with Eosinophilic Granulomatosis with Polyangiitis That Responded to Glucocorticoid Therapy (2023)
  • Author(s): Ozaka Sotaro、Kodera Takamoto、Tsutsumi Koshiro、Fukuda Masahide、Iwao Masao、Hirashita Yuka、Takahashi Haruhiko、Fukuda Kensuke、Okamoto Kazuhisa、Arakawa Mie、Ogawa Ryo、Endo Mizuki、Mizukami Kazuhiro、Okimoto Tadayoshi、Kamiyama Naganori、Kodama Masaaki、Kobayashi Takashi、Murakami Kazunari
  • Journal Title: Internal Medicine Volume: 62 Issue: 10 Pages: 1501-1506
  • DOI: 10.2169/internalmedicine.0437-22
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2023-05-15
  • Peer Reviewed / Open Access
• [Journal Article] CCL20/CCR6 chemokine signaling is not essential for pathogenesis in an experimental autoimmune encephalomyelitis mouse model of multiple sclerosis (2023)
  • Author(s): Sachi Nozomi、Kamiyama Naganori、Saechue Benjawan、Ozaka Sotaro、Dewayani Astri、Ariki Shimpei、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Ekronarongchai Supanuch、Kobayashi Takashi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 641 Pages: 123-131
  • DOI: 10.1016/j.bbrc.2022.11.088
  • Peer Reviewed
• [Journal Article] GM-CSF-producing CCR2+ CCR6+ Th17 cells are pathogenic in dextran sodium sulfate-induced colitis model in mice (2023)
  • Author(s): Ariki Shimpei、Ozaka Sotaro、Sachi Nozomi、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Ekronarongchai Supanuch、Mizukami Kazuhiro、Kamiyama Naganori、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Genes to Cells Volume: 28 Issue: 4 Pages: 267-276
  • DOI: 10.1111/gtc.13008
  • Peer Reviewed
• [Journal Article] Differences in clinical features and morphology between differentiated and undifferentiated gastric cancer after Helicobacter pylori eradication (2023)
  • Author(s): Kodama M, Mizukami K, Hirashita Y, Okimoto T, Wada Y, Fukuda M, Ozaka S, Kudo Y, Ito K, Ogawa R, Okamoto K, Fukuda K, Murakami K
  • Journal Title: PLos One Volume: 18 Issue: 3 Pages: e0282341-e0282341
  • DOI: 10.1371/journal.pone.0282341
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive lipidomics of lupus-prone mice using LC-MS/MS identifies the reduction of palmitoylethanolamide that suppresses TLR9-mediated inflammation. (2022)
  • Author(s): Ozaki T, Kamiyama N, Saechue B, Soga Y, Gotoh R, Nakayama T, Fukuda C, Dewayani A, Chalalai T, Ariki S, Ozaka S, Sonoda A, Hirose H, Gendo Y, Noguchi K, Sachi N, Hidano S, Maeshima K, Gotoh K, Masaki T, Ishii K, Osada Y, Shibata H, Kobayashi T
  • Journal Title: Genes Cells. Volume: 27 Issue: 7 Pages: 493-504
  • DOI: 10.1111/gtc.12944
  • Peer Reviewed / Open Access
• [Journal Article] TRAF6 signaling pathway in T cells regulates anti-tumor immunity through the activation of tumor specific Th9 cells and CTLs (2022)
  • Author(s): Dewayani Astri、Kamiyama Naganori、Sachi Nozomi、Ozaka Sotaro、Saechue Benjawan、Ariki Shimpei、Goto Mizuki、Chalalai Thanyakorn、Soga Yasuhiro、Fukuda Chiaki、Kagoshima Yomei、Maekawa Yoichi、Kobayashi Takashi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 613 Pages: 26-33
  • DOI: 10.1016/j.bbrc.2022.04.125
  • Peer Reviewed
• [Journal Article] Saireito, a Japanese herbal medicine, alleviates leaky gut associated with antibiotic-induced dysbiosis in mice (2022)
  • Author(s): Ozaka S、Sonoda A、Ariki S、Minata M、Kamiyama N、Hidano S、Sachi N、Ito K、Kudo Y、Dewayani A、Chalalai T、Ozaki T、Soga Y、Fukuda C、Mizukami K、Ishizawa S、Nishiyama M、Fujitsuka N、Mogami S、Kubota K、Murakami K、Kobayashi T
  • Journal Title: PLOS ONE Volume: 17 Issue: 6 Pages: e0269698-e0269698
  • DOI: 10.1371/journal.pone.0269698
  • Peer Reviewed / Open Access
• [Journal Article] Potential association of eEF1A dimethylation at lysine 55 in the basal area of Helicobacter pylori-eradicated gastric mucosa with the risk of gastric cancer: a retrospective observational study (2022)
  • Author(s): Hirashita Yuka、Fukuda Masahide、Kodama Masaaki、Tsukamoto Yoshiyuki、Okimoto Tadayoshi、Mizukami Kazuhiro、Kawahara Yoshinari、Wada Yasuhiro、Ozaka Sotaro、Togo Kazumi、Kinoshita Keisuke、Fuchino Takafumi、Fukuda Kensuke、Okamoto Kazuhisa、Ogawa Ryo、Matsunari Osamu、Honda Koichi、Murakami Kazunari
  • Journal Title: BMC Gastroenterology Volume: 22 Issue: 1 Pages: 490-490
  • DOI: 10.1186/s12876-022-02521-5
  • Peer Reviewed / Open Access
• [Journal Article] COVID-19 vaccination and liver disease (2022)
  • Author(s): Ozaka Sotaro、Kobayashi Takashi、Mizukami Kazuhiro、Murakami Kazunari
  • Journal Title: World Journal of Gastroenterology Volume: 28 Issue: 48 Pages: 6791-6810
  • DOI: 10.3748/wjg.v28.i48.6791
  • Peer Reviewed / Open Access
• [Journal Article] Acute pancreatitis soon after COVID-19 vaccination (2022)
  • Author(s): Ozaka Sotaro、Kodera Takamoto、Ariki Shimpei、Kobayashi Takashi、Murakami Kazunari
  • Journal Title: Medicine Volume: 101 Issue: 2 Pages: e28471-e28471
  • DOI: 10.1097/md.0000000000028471
  • Peer Reviewed / Open Access
• [Journal Article] Endoscopic Removal of Multiple Ingested Cylindrical Batteries (2022)
  • Author(s): Ozaka Sotaro、Inoue Kunimitsu、Tasaki Takako、Ono Hideki、Murakami Kazunari
  • Journal Title: Cureus Volume: -
  • DOI: 10.7759/cureus.21789
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of the Prevalence and Species of <i>Anisakis</i> nematode in Sekisaba, <i>Scomber japonicus</i> Caught in Coastal Waters off Saganoseki, Oita in Japan (2021)
  • Author(s): Hidano Shinya、Mizukami Kazuhiro、Yahiro Takaaki、Shirakami Kohei、Ito Hideyuki、Ozaka Sotaro、Ariki Shimpei、Saechue Benjawan、Dewayani Astri、Chalalai Thanyakorn、Soga Yasuhiro、Goto Mizuki、Sonoda Akira、Ozaki Takashi、Sachi Nozomi、Kamiyama Naganori、Nishizono Akira、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 74 Issue: 5 Pages: 387-391
  • DOI: 10.7883/yoken.JJID.2020.859
  • NAID: 130008090913
  • ISSN: 1344-6304, 1884-2836
  • Year and Date: 2021-09-30
  • Peer Reviewed / Open Access
• [Journal Article] Comment on “A case of acute necrotising pancreatitis following the second dose of Pfizer‐BioNTech COVID‐19 mRNA vaccine” (2021)
  • Author(s): Ozaka Sotaro、Kobayashi Takashi、Murakami Kazunari
  • Journal Title: British Journal of Clinical Pharmacology Volume: 88 Issue: 5 Pages: 2449-2449
  • DOI: 10.1111/bcp.15159
  • Peer Reviewed / Open Access
• [Journal Article] Protease inhibitory activity of secretory leukocyte protease inhibitor ameliorates murine experimental colitis by protecting the intestinal epithelial barrier (2021)
  • Author(s): Ozaka S, Sonoda A, Ariki S, Kamiyama N, Hidano S, Sachi N, Ito K, Kudo Y, Minata M, Saechue B, Dewayani A, Chalalai T, Soga Y, Takahashi Y, Fukuda C, Mizukami K, Okumura R, Kayama H, Murakami K, Takeda K, Kobayashi T.
  • Journal Title: Genes to Cells Volume: 26 Issue: 10 Pages: 807-822
  • DOI: 10.1111/gtc.12888
  • Peer Reviewed / Open Access
• [Journal Article] Monomorphic epitheliotropic intestinal T-cell lymphoma presenting as melena with long-term survival: A case report and review of literature (2021)
  • Author(s): Ozaka Sotaro、Inoue Kunimitsu、Okajima Tomoya、Tasaki Takako、Ariki Shimpei、Ono Hideki、Ando Takeaki、Daa Tsutomu、Murakami Kazunari
  • Journal Title: World Journal of Gastroenterology Volume: 27 Issue: 38 Pages: 6501-6510
  • DOI: 10.3748/wjg.v27.i38.6501
  • Peer Reviewed / Open Access
• [Journal Article] Rectal varix treated with endoscopic cyanoacrylate injection therapy (2021)
  • Author(s): Ozaka Sotaro、Gotoh Yasuhiko、Honda Shunichiro、Iwao Toshiaki、Kawahara Yoshinari、Kinoshita Keisuke、Nakaya Tomohisa、Noguchi Chishio、Kagawa Koichi、Murakami Kazunari
  • Journal Title: Clinical Journal of Gastroenterology Volume: - Issue: 3 Pages: 791-795
  • DOI: 10.1007/s12328-020-01305-2
  • Peer Reviewed
• [Journal Article] Development of a portable reverse transcription loop‐mediated isothermal amplification system to detect the E1 region of Chikungunya virus in a cost‐effective manner (2020)
  • Author(s): Saechue Benjawan、Kamiyama Naganori、Fukuda Chiaki、Watanabe Kei、Soga Yasuhiro、Goto Mizuki、Dewayani Astri、Ariki Shimpei、Hirose Haruna、Ozaka Sotaro、Sachi Nozomi、Hidano Shinya、Shirin Tahmina、Mondal Dinesh、Murakami Kazunari、Kobayashi Takashi
  • Journal Title: Genes to Cells Volume: 9 Issue: 9 Pages: 615-625
  • DOI: 10.1111/gtc.12797
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 抗菌薬で生じるリーキーガットに対する柴苓湯の保護効果に関する基礎研究 (2023)
  • Author(s): 小坂聡太郎, 神山長慶, 佐知望美, 堤康志郎, 髙橋晴彦, 福田健介, 小川竜, 水上一弘, 小林隆志, 村上和成
  • Organizer: 第19回日本消化管学会総会学術総会
• [Presentation] Dysbiosisに起因するLeaky gutモデルに対する柴苓湯の有効性 (2022)
  • Author(s): 小坂 聡太郎, 有木 晋平, 水上 一弘, 神山 長慶, 福田 健介, 岡本 和久, 小川 竜, 沖本 忠義, 小林 隆志, 村上 和成
  • Organizer: 第30回日本消化器関連学会週間 (JDDW 2022)
• [Presentation] 分泌型好中球プロテアーゼインヒビターは腸管上皮バリアを保護することでマウス実験腸炎を軽減する (2022)
  • Author(s): 小坂聡太郎, 有木晋平, 神山長慶, 鹿子嶋洋明, 佐知望美, Astri Dewayani, Thanyakorn Chalalai, 水上一弘, 竹田潔, 村上和成, 小林隆志
  • Organizer: 第51回日本免疫学会学術集会
• [Presentation] マウス腸炎モデルを用いた大建中湯のSLPI発現を介した腸管保護作用の解析 (2021)
  • Author(s): 小坂聡太郎, 皆田美月, 有木晋平, 水上一弘, 神山長慶, 小林隆志, 村上和成.
  • Organizer: 第29回日本消化器関連学会週間 (JDDW 2021)
• [Presentation] 柴苓湯はマウスにおいて抗菌薬によるdysbiosisに起因するLeaky gutを軽減する (2021)
  • Author(s): 小坂聡太郎, 有木晋平, 皆田美月, 神山長慶，佐知望美，Benjawan Saechue，Astri Dewayani, Thanyakorn Chalalai, 曽我泰裕, 福田千瑛, 水上一弘, 村上和成, 小林隆志
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 大建中湯のSLPI発現増強を介した腸管保護作用の解析 (2021)
  • Author(s): 皆田美月, 小坂聡太郎, 有木晋平, 神山長慶, 佐知望美, Benjawan Saechue, Astri Dewayani, Thanyakorn Chalalai, 曽我泰裕, 福田千瑛, 水上一弘, 村上和成, 小林隆志
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 上部消化管出血を契機に診断され長期生存している単形性上皮向性腸管T細胞リンパ腫の1例 (2021)
  • Author(s): 小坂聡太郎, 井上邦光, 岡嶋智也, 田崎貴子, 有木晋平, 小野英樹, 村上和成
  • Organizer: 第111回日本消化器内視鏡学会九州支部例会
• [Presentation] Physiological relevance of TRAF6 signaling in dendritic cells in controlling C. rodentium infection (2021)
  • Author(s): Chalalai T., Kamiyama N., Saechue B., Dewayani A., Ariki S., Ozaka S., Sachi N., Fukuda C., Soga Y., Okumura R., Kayama H., Takeda K., Kobayashi T
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] Establishment of a reporter assay system to quantity TRAF6 mediated inflammatory signals and search for novel anti-inflammatory. (2021)
  • Author(s): Soga Y., Kamiyama N., Saechue B., Dewayani A., Hidano S., Sachi N., Ozaka S., Ariki S., Goto M., Kobayashi T.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] CCL20欠損マウスを用いた多発性硬化症の発症におけるCCL20の機能解析. (2021)
  • Author(s): 佐知望美，神山長慶，Benjawan Saechue，小坂聡太郎，Astri Dewayani，有木晋平，Thanyakorn Chalalai，曽我泰裕，福田千瑛，小林隆志.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] TRAF6 Regulates Th9 Cells and Cytotoxic T Lymphocytes in Tumor Immunity. (2021)
  • Author(s): Dewayani A., Kamiyama N., Hidano S., Sachi N., Ozaka S., Ariki S., Saechue B., Soga Y., Chalalai T., Fukuda C., Kobayashi T.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 一回感染性ジカウイルス様粒子（SRIPｓ）を用いた宿主細胞侵入に関わるprME領域のアミノ酸の同定. (2021)
  • Author(s): 福田千瑛，神山長慶，Benjawan Saechue，Astri Dewayani，Thanyakorn Chalalai，小坂聡太郎，有木晋平，曽我泰裕，皆田美月，佐知望美，飛彈野真也，鈴木亮介，小林隆志.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] TRAF6 promotes the migration of Th17 cells into the CNS by regulating CCR6 expression in experimental autoimmune encephalomyelitis mice. (2021)
  • Author(s): Kamiyama N., Saechue B., Dewayani A., Thanyakorn C., Ozaka S., Ariki S., Soga Y., Goto M., Sachi N., Hidano S., Kobayashi T.
  • Organizer: 第44回日本分子生物学会年会
• [Presentation] 抗生物質起因性大腸炎マウスモデルに対する柴苓湯の有効性の解明 (2020)
  • Author(s): 小坂聡太郎, 園田光, 有木晋平, 水上一弘, 村上和成
  • Organizer: 第106回日本消化器病学会総会
• [Presentation] DSS腸炎モデルを用いたSLPIの結腸における生理的機能の解析 (2020)
  • Author(s): 小坂聡太郎, 有木晋平, 園田光, 神山長慶, 飛彈野真也, 水上一弘, 小林隆志, 村上和成
  • Organizer: 第28回 JDDW 日本消化器関連学会
• [Presentation] 内視鏡的シアノアクリレート注入療法が奏功した直腸静脈瘤の1例 (2020)
  • Author(s): 小坂聡太郎, 本田俊一郎, 川原義成, 仲谷朋久, 野口地塩, 後藤康彦, 香川浩一, 村上和成
  • Organizer: 第110回日本消化器内視鏡学会九州支部例会
• [Book] 国民の栄養白書2021年度版 (2021)
  • Author(s): 小坂聡太郎, 小林隆志
  • Total Pages: 204
  • Publisher: 日本医療企画
  • ISBN: 9784867290811
• [Book] 消化性潰瘍診療ガイドライン2020改訂第3版 (2020)
  • Author(s): 小池和彦，渡辺純夫，佐藤貴一，鎌田智有，伊藤俊之，伊藤公訓，岩本淳一，沖本忠義，小坂聡太郎 他
  • Total Pages: 199
  • Publisher: 南江堂