18‑&alpha;‑glycyrrhetinic acid induces apoptosis in gingival fibroblasts exposed to phenytoin

Takeuchi,Reiri; Nomura,Takatoshi; Yaguchi,Manabu; Taguchi,Chieko; Suzuki,Itaru; Suzuki,Haruka; Matsumoto,Hiroko; Okada,Yuichiro; Arikawa,Kazumune; Nomoto,Takato; Hiratsuka,Koichi

doi:10.3892/etm.2024.12586

18‑α‑glycyrrhetinic acid induces apoptosis in gingival fibroblasts exposed to phenytoin

Authors:
- Reiri Takeuchi
- Takatoshi Nomura
- Manabu Yaguchi
- Chieko Taguchi
- Itaru Suzuki
- Haruka Suzuki
- Hiroko Matsumoto
- Yuichiro Okada
- Kazumune Arikawa
- Takato Nomoto
- Koichi Hiratsuka
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Affiliations: Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271‑8587, Japan, Department of Special Needs Dentistry, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271‑8587, Japan, Department of Preventive and Public Oral Health, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271‑8587, Japan, Department of Pharmacology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271‑8587, Japan, Department of Histology, Nihon University School of Dentistry at Matsudo, Matsudo, Chiba 271‑8587, Japan
Published online on: May 22, 2024 https://doi.org/10.3892/etm.2024.12586
Article Number: 297
Copyright : © Takeuchi et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].

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Abstract

Phenytoin (PHT)‑induced gingival overgrowth is caused by the increased proliferation and reduced apoptosis of gingival fibroblasts in inflammatory gingiva. Licorice has long been used as a component of therapeutic preparations. It inhibits cell proliferation, induces cell apoptosis and has anti‑inflammatory effects. 18‑α‑glycyrrhetinic acid (18α‑GA), the active compound in licorice, promotes apoptosis in various types of cells. The present study determined whether 18α‑GA affects apoptosis in gingival fibroblasts exposed to PHT. The present study aimed to establish a basis for the therapeutic application of 18α‑GA to treat the gingival overgrowth induced by PHT. Human gingival ﬁbroblasts from healthy donors were cultured to semi‑conﬂuence and then stimulated in serum‑free DMEM containing PHT with or without 18α‑GA for subsequent experiments. Apoptotic cells were detected by ELISA. Analysis of the distribution of cell cycle phases and the apoptotic cell population was performed by flow cytometry. The expression levels of mRNAs and proteins of apoptotic regulators were measured using reverse transcription‑quantitative PCR and western blotting, respectively. Caspase (CASP) activities were assessed by an ELISA. Treatment with 18α‑GA markedly increased the number of apoptotic cells, reduced BCL2 mRNA expression, increased CASP2 and receptor (TNFRSF)‑interacting serine‑threonine kinase 1 (RIPK1) domain containing adaptor with death domain, Fas (TNFRSF6)‑associated via death domain, RIPK1, tumor necrosis factor receptor superfamily; member 1A, TNF receptor‑associated factor 2, CASP2, CASP3 and CASP9 mRNA expression, and also upregulated the protein expression levels and activities of caspase‑2, caspase‑3 and caspase‑9. These results demonstrated that 18α‑GA induced apoptosis through the activation of the Fas and TNF pathways in the death receptor signaling pathway in gingival fibroblasts treated with PHT. 18α‑GA exhibited therapeutic potential for the treatment of PHT‑induced gingival overgrowth.

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