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Cytoprotective effects of Hangekobokuto against corticosterone-induced cell death in HT22 cells

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Abstract

The hypothalamic–pituitary–adrenal (HPA) system plays an important role in stress response. Chronic stress is thought to induce neuronal damage and contribute to the pathogenesis of psychiatric disorders by causing dysfunction of the HPA system and promoting the production and release of glucocorticoids, including corticosterone and cortisol. Several clinical studies have demonstrated the efficacy of herbal medicines in treating psychiatric disorders; however, their effects on corticosterone-induced neuronal cell death remain unclear. Here, we used HT22 cells to evaluate the neuroprotective potential of herbal medicines used in neuropsychiatry against corticosterone-induced hippocampal neuronal cell death. Cell death was assessed using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction and Live/Dead assays. Hangekobokuto, Kamikihito, Saikokaryukotsuboreito, Kamishoyosan, and Yokukansan were supplied in the form of water-extracted dried powders. Exposure of HT22 cells to ≥ 100 μM corticosterone decreased MTT values. Exposure to 500 μM corticosterone alone reduced MTT values to 18%, while exposure to 10 μM Mifepristone (RU486)—a glucocorticoid receptor antagonist—restored values to 36%. Corticosterone-induced cell death was partially suppressed by treatment with RU486. At 100 μg/mL, Hangekobokuto significantly suppressed the decrease in MTT values (15–32%) and increase in the percentage of ethidium homodimer-1-positive dead cells caused by corticosterone exposure (78–36%), indicating an inhibitory effect on cell death. By contrast, Kamikihito, Saikokaryukotsuboreito, Kamishoyosan, and Yokukansan did not affect corticosterone-induced cell death. Therefore, our results suggest that Hangekobokuto may ameliorate the onset and progression of psychiatric disorders by suppressing neurological disorders associated with increased levels of glucocorticoids.

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Acknowledgements

We thank all members of our laboratory, especially T. Murai, K. Hayakawa, A. Takahashi, and K. Tsuruta, for their excellent technical assistance.

Funding

This work was funded in part by JSPS KAKENHI Grant Number 21K06620 (HM and YK), a grant to encourage and promote research projects at the School of Pharmacy, Nihon University (2020–2021, HM), and a Nihon University Research Grant for 2022–2023 (YK). The funding bodies had no role in the design of the study or writing of the manuscript.

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Authors and Affiliations

  1. Laboratory of Pharmacology, School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi-Shi, Chiba, 274-8555, Japan

    Hiroko Miyagishi, Ami Joyama, Hiroshi Nango, Koume Nagayama & Yasuhiro Kosuge

  2. Department of Pharmacology, School of Pharmacy, International University of Health and Welfare, 2600-1 Kitakanemaru, Ohtawara, Tochigi, 324-8501, Japan

    Hiroko Miyagishi & Minoru Tsuji

  3. Department of Pharmacology, School of Pharmacy at Fukuoka, International University of Health and Welfare, 137-1 Enokizu, Okawa, Fukuoka, 831-8501, Japan

    Hiroshi Takeda

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  1. Hiroko Miyagishi
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  2. Ami Joyama
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  3. Hiroshi Nango
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  4. Koume Nagayama
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  5. Minoru Tsuji
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  7. Yasuhiro Kosuge
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Contributions

HM and YK designed the study. HM and AJ performed the experiments. HM, AJ, HN and KN analyzed and interpreted the data. MT and HT contributed to the preparation of the reagents and materials. HM and YK wrote the manuscript and YK, HN, MT and HT reviewed the manuscript. All the authors have read and approved the final manuscript.

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Correspondence to Hiroko Miyagishi or Yasuhiro Kosuge.

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Miyagishi, H., Joyama, A., Nango, H. et al. Cytoprotective effects of Hangekobokuto against corticosterone-induced cell death in HT22 cells. J Nat Med 78, 255–265 (2024). https://doi.org/10.1007/s11418-023-01766-y

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