The effect of intermittent hypoxia on the expression of insulin resistance-related genes in muscle cells

• Project/Area Number: 21K16344
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 54040:Metabolism and endocrinology-related
• Research Institution: Nara Medical University
• Principal Investigator: Shobatake Ryogo 奈良県立医科大学, 医学部, 博士研究員 (40771027)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2022: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2021: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 間歇的低酸素 / インスリン抵抗性 / ミオカイン / 睡眠時無呼吸症候群 / 2型糖尿病 / 糖尿病 / Intermittent hypoxia / myokines / IL-8 / osteonectin / myonectin / OCT1 / NRF2 / 骨格筋

Outline of Research at the Start

睡眠時無呼吸症候群 (Sleep apnea syndrome; SAS) 患者は睡眠中に繰り返す無呼吸により、全身の組織、細胞は間歇的低酸素 (Intermittent hypoxia; IH)に曝露される。SASの重症化はインスリン抵抗性との関連が指摘されている。骨格筋はグルコースの取り込みにとって重要な代謝組織である。今回、マウスC2C12細胞とヒトRD細胞を用いてIHがグルコースの取り込みや利用に関わる遺伝子と筋細胞で発現・分泌されるインスリン抵抗性に関わる生理活性因子の遺伝子発現に与える影響とその分子機序について検討する。

Outline of Final Research Achievements

Sleep apnea syndrome is characterized by recurrent episodes of oxygen desaturation and reoxygenation (intermittent hypoxia [IH]) and is a risk factor for insulin resistance/Type 2 diabetes. We exposed human RD and mouse C2C12 muscle cells to normoxia or IH and measured their mRNA levels by real-time RT-PCR. We found that IH significantly increased the mRNA and protein levels of muscle-derived insulin resistance-factors (myokines) such as IL-8, osteonectin (ON), and myonectin (MN) in muscle cells. Deletion analyses revealed that the regions -152 to -151 in IL-8, -105 to -99 in ON, and - 3741 to -3738 in MN promoters were responsible for the activation by IH in RD cells. The promoters contain consensus transcription factor binding sequences for OCT1 in IL-8 and MN promoters, and for NRF2 in ON promoter, respectively. The introduction of siRNA for OCT1 abolished the IH-induced expression(s) of IL-8 and MN and siRNA for NRF2 abolished the IH-induced expression of ON.

Academic Significance and Societal Importance of the Research Achievements

睡眠時無呼吸症候群の主病態である間歇的低酸素 （IH）が、インスリン抵抗性に関連するミオカイン遺伝子の発現に与える影響を検討した。IHにより筋細胞においてIL-8、osteonectin (ON)、myonectin (MN) 遺伝子発現が転写レベルで亢進し、IL-8、MNはOCT1、ONはNRF2を転写因子として必要とすることを示した。SAS患者では、IL-8、ON、MNの遺伝子発現増加が、筋肉組織において炎症性表現型を誘導し、インスリン抵抗性に寄与していると考えられた。

Research Products

• [Journal Article] Autoimmune Encephalitis Associated with Anti-N-methyl-D-aspartate Receptor and Anti-Hu Antibodies Successfully Treated with Carboplatin and Etoposide for Small-cell Lung Cancer (2023)
  • Author(s): Shobatake Ryogo、Kumazawa Aya、Koyama Noriko、Takahashi Nobuyuki
  • Journal Title: Internal Medicine Volume: 62 Issue: 3 Pages: 469-474
  • DOI: 10.2169/internalmedicine.9707-22
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2023-02-01
  • Peer Reviewed / Open Access
• [Journal Article] Intermittent Hypoxia Increased the Expression of DBH and PNMT in Neuroblastoma Cells via MicroRNA-375-Mediated Mechanism (2022)
  • Author(s): Takasawa Shin, Shobatake Ryogo, Takeda Yoshinori, Uchiyama Tomoko, Yamauchi Akiyo, Makino Mai, Sakuramoto-Tsuchida Sumiyo, Asai Keito, Ota Hiroyo, Itaya-Hironaka Asako
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 11 Pages: 5868-5868
  • DOI: 10.3390/ijms23115868
  • Peer Reviewed / Open Access
• [Journal Article] Downregulation of the Cd38-Cyclic ADP-Ribose Signaling in Cardiomyocytes by Intermittent Hypoxia via Pten Upregulation (2022)
  • Author(s): Takasawa Shin, Makino Mai, Uchiyama Tomoko, Yamauchi Akiyo, Sakuramoto-Tsuchida Sumiyo, Itaya-Hironaka Asako, Takeda Yoshinori, Asai Keito, Shobatake Ryogo, Ota Hiroyo
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 15 Pages: 8782-8782
  • DOI: 10.3390/ijms23158782
  • Peer Reviewed / Open Access
• [Journal Article] Age and composition of the thrombus retrieved by mechanical thrombectomy from patients with acute ischemic stroke are associated with revascularization and clinical outcomes (2022)
  • Author(s): Shimizu Hisao、Hatakeyama Kinta、Saito Kozue、Shobatake Ryogo、Takahashi Nobuyuki、Deguchi Jun、Tokunaga Hidemori、Shimada Keiji、Nakagawa Ichiro、Myochin Kaoru、Sakai Kazuya、Kubo Masayuki、Yamashita Atsushi、Obayashi Chiho、Sugie Kazuma、Matsumoto Masanori
  • Journal Title: Thrombosis Research Volume: 219 Pages: 60-69
  • DOI: 10.1016/j.thromres.2022.09.004
  • Peer Reviewed
• [Journal Article] Upregulation of Reg IV and Hgf mRNAs by Intermittent Hypoxia via Downregulation of microRNA-499 in Cardiomyocytes (2022)
  • Author(s): Takasawa Shin, Itaya-Hironaka Asako, Makino Mai, Yamauchi Akiyo, Sakuramoto-Tsuchida Sumiyo, Uchiyama Tomoko, Shobatake Ryogo, Takeda Yoshinori, Ota Hiroyo
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 20 Pages: 12414-12414
  • DOI: 10.3390/ijms232012414
  • Peer Reviewed / Open Access
• [Journal Article] The Impact of Intermittent Hypoxia on Metabolism and Cognition (2022)
  • Author(s): Shobatake Ryogo、Ota Hiroyo、Takahashi Nobuyuki、Ueno Satoshi、Sugie Kazuma、Takasawa Shin
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 21 Pages: 12957-12957
  • DOI: 10.3390/ijms232112957
  • Peer Reviewed / Open Access
• [Journal Article] Upregulation of IL‐8, osteonectin, and myonectin mRNAs by intermittent hypoxia via OCT1‐ and NRF2‐mediated mechanisms in skeletal muscle cells (2022)
  • Author(s): Takasawa Shin, Shobatake Ryogo, Itaya‐Hironaka Asako, Makino Mai, Uchiyama Tomoko, Sakuramoto‐Tsuchida Sumiyo, Takeda Yoshinori, Ota Hiroyo, Yamauchi Akiyo
  • Journal Title: Journal of Cellular and Molecular Medicine Volume: 26 Issue: 24 Pages: 6019-6031
  • DOI: 10.1111/jcmm.17618
  • Peer Reviewed / Open Access
• [Journal Article] Anorexigenic Effects of Intermittent Hypoxia on the Gut-Brain Axis in Sleep Apnea Syndrome (2021)
  • Author(s): Shobatake Ryogo、Ota Hiroyo、Takahashi Nobuyuki、Ueno Satoshi、Sugie Kazuma、Takasawa Shin
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 1 Pages: 364-364
  • DOI: 10.3390/ijms23010364
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Intermittent Hypoxia and Appetite Regulation in the Gut-Brain Axis (2023)
  • Author(s): Ryogo Shobatake, Nobuyuki Takahashi, Satoshi Ueno, Kazuma Sugie, Shin Takasawa
  • Organizer: 13th Annual World Congress of NeuroTalk-2023
  • Invited
• [Presentation] 間歇的低酸素は血管内皮細胞のICAM-1, ESM1 mRNAを増加させる (2023)
  • Author(s): 高沢 伸, 牧野 舞, 山内 晶世, 土田 澄代, 廣田 梨奈, 藤井 龍星, 朝井 啓斗, 竹田善紀, 内山 智子,正畠 良悟, 太田 浩世
  • Organizer: 第66回日本糖尿病学会年次学術集会
• [Presentation] Intermittent Hypoxia Increased the Expression of Intercellular Adhesion Molecule-1 and Endothelial Cell-Specific Molecule 1 in Vascular Endothelial Cells via the Downregulation of MicroRNA-181a1 (2023)
  • Author(s): Shin Takasawa, Mai Makino, Akiyo Yamauchi, Sumiyo Sakuramoto-Tsuchida, Rina Hirota, Ryusei Fujii, Keito Asai, Yoshinori Takeda, Tomoko Uchiyama, Ryogo Shobatake, Hiroyo Ota
  • Organizer: 83rd ADA Scientific Sessions
  • Int'l Joint Research
• [Presentation] Gene expression of cardiomyocytes by intermittent hypoxia: the Cd38-cyclic ADP-ribose (cADPR) signaling for functioning and the Reg-Reg receptor system for regeneration (2023)
  • Author(s): Shin Takasawa, Mai Makino, Tomoko Uchiyama, Akiyo Yamauchi, Sumiyo Sakuramoto-Tschida, Asako Itaya-Hironaka, Yoshinori Takeda, Keito Asai, Ryogo Shobatake, Hiroyo Ota
  • Organizer: 4th Edition of World Congress on ENDOCRINOLOGY, DIABETES AND METABOLISM
  • Int'l Joint Research
• [Presentation] Intermittent hypoxia increased the expression of DBH and PNMT in neuronal cells via microRNA-mediated mechanism. (2022)
  • Author(s): Takasawa, S., Shobatake, R., Takeda, Y., Uchiyama, T., Yamauchi, A., Makino, M., Asai, K. and Ota, H.
  • Organizer: 82nd American Diabetes Association Scientific Sessions, June 3-7, 2022, New Orleans, LA, Hybrid Diabetes 71 (Suppl. 1), 1317-P (2022).
  • Int'l Joint Research
• [Presentation] Attenuation of the CD38-cADPR signal system in cardiomyocytes by intermittent hypoxia via upregulation of Pten (2022)
  • Author(s): Shin Takasawa, Mai Makino, Tomoko Uchiyama, Akiyo Yamauchi, Sumiyo Sakuramoto-Tsuchida, Asako Itaya-Hironaka, Yoshinori Takeda, Keito Asai, Ryogo Shobatake, Hiroyo Ota
  • Organizer: International Diabetes Federation (IDF)
• [Presentation] 間歇的低酸素は神経細胞においてカテコールアミン合成酵素mRNAを増加させる (2022)
  • Author(s): 高沢 伸, 正畠 良悟, 竹田 善紀,内山智子, 山内 晶世, 牧野 舞,土田 澄代, 朝井啓斗, 太田 浩世, 広中 安佐子
  • Organizer: 第65回日本糖尿病学会年次学術集会：一般演題 口演（a90868）
• [Presentation] Intermittent hypoxia increased the expression of DBH and PNMT in neuronal cells via microRNA-mediated mechanism. (2022)
  • Author(s): Takasawa, S., Shobatake, R., Takeda, Y., Uchiyama, T., Yamauchi, A., Makino, M., Asai, K. and Ota, H.
  • Organizer: 82nd American Diabetes Association Scientific Sessions
  • Int'l Joint Research
• [Presentation] Intermittent hypoxia up-regulates gene expressions of anorexigenic peptides in neuronal and enteroendocrine cells by different mechanisms (2021)
  • Author(s): Ryogo Shobatake
  • Organizer: ESMED Congress 2021
  • Int'l Joint Research / Invited