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Modafinil exerts anti-inflammatory and anti-fibrotic effects by upregulating adenosine A2A and A2B receptors

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Abstract

Adenosine receptor (AR) suppresses inflammation and fibrosis by activating cyclic adenosine monophosphate (cAMP) signaling. We investigated whether altered AR expression contributes to the development of fibrotic diseases and whether A2AAR and A2BAR upregulation inhibits fibrotic responses. Primary human lung fibroblasts (HLFs) from normal (NHLFs) or patients with idiopathic pulmonary fibrosis (DHLF) were used for in vitro testing. Murine models of fibrotic liver or pulmonary disease were developed by injecting thioacetamide intraperitoneally, by feeding a high-fat diet, or by intratracheal instillation of bleomycin. Modafinil, which activates cAMP signaling via A2AAR and A2BAR, was administered orally. The protein amounts of A2AAR, A2BAR, and exchange protein directly activated by cAMP (Epac) were reduced, while collagen and α-smooth muscle actin (α-SMA) were elevated in DHLFs compared to NHLFs. In liver or lung tissue from murine models of fibrotic diseases, A2AAR and A2BAR were downregulated, but A1AR and A3AR were not. Epac amounts decreased, and amounts of collagen, α-SMA, KCa2.3, and KCa3.1 increased compared to the control. Modafinil restored the amounts of A2AAR, A2BAR, and Epac, and reduced collagen, α-SMA, KCa2.3, and KCa3.1 in murine models of fibrotic diseases. Transforming growth factor-β reduced the amounts of A2AAR, A2BAR, and Epac, and elevated collagen, α-SMA, KCa2.3, and KCa3.1 in NHLFs; however, these alterations were inhibited by modafinil. Our investigation revealed that A2AAR and A2BAR downregulation induced liver and lung fibrotic diseases while upregulation attenuated fibrotic responses, suggesting that A2AAR and A2BAR-upregulating agents, such as modafinil, may serve as novel therapies for fibrotic diseases.

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Data availability

The datasets and materials generated and analysed during the current study are available from the corresponding authors on reasonable request.

Abbreviations

α-SMA:

α-smooth muscle actin

AC:

Adenylyl cyclase

AR:

Adenosine receptor

Bleomycin:

BLM

cAMP:

Cyclic adenosine monophosphate

CDAHFD:

Choline-deficient, L-amino-acid-defined, high-fat diet with 0.1% methionine

Col:

Collagen

DMSO:

Dimethyl sulfoxide

Epac:

Exchange protein directly activated by the cAMP

HSC:

Hepatic stellate cell

H&E:

Hematoxylin and eosin

IL:

Interleukin

IPF:

Idiopathic pulmonary fibrosis

MF:

Modafinil

MF-S:

(S)-Isomer of modafinil

MMP9:

Matrix metalloproteinase-9

MRS:

MRS 1754

NECA:

5’-N-ethylcarboxamidoadenosine

PCR:

Polymerase chain reaction

P1NP:

Procollagen type1 N-terminal propeptide

A2 :

AR-siRs siRNA against A2AAR + siRNA against A2BAR

TAA:

Thioacetamide

TIMP:

Tissue inhibitor of metalloproteinases

TGFβ:

Transforming growth factor-β

TNFα:

Tumor necrosis factor-α

ZM:

ZM 241,385

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Funding

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (NRF-2022R1A2C1007823), NRF grant funded by the Korea government (MSIT) (NRF-2022R1A2C1092484), Ewha Womans University Research Grant of 2022 (1-2022-1108-001-1) and intramural research promotion grants from Ewha Womans University, School of Medicine (RP-2020).

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Author notes

  1. KC Kim, S Choi, and SH Suh contributed equally to this publication and share therefore the corresponding authorship.

Authors and Affiliations

  1. Department of Physiology, College of Medicine, Ewha Womans University, 25 Magokdong-ro 2-gil, Gangseo-gu, Seoul, 07084, Republic of Korea

    Haiyan Li, Ji Aee Kim, Seong-Eun Jo, Huisu Lee, Shinkyu Choi & Suk Hyo Suh

  2. Department of Thoracic & Cardiovascular Surgery, College of Medicine, Ewha Womans University, 25 Magokdong-ro 2-gil, Gangseo-gu, Seoul, 07084, Republic of Korea

    Kwan-Chang Kim

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Contributions

Haiyan Li, Jee Aee Kim, Seong-Eun Jo, Huisu Lee: performed the experiments; Haiyan Li and Suk Hyo Suh: wrote the manuscipt; Kwan-Chang Kim, Shinkyu Choi, and Suk Hyo Suh: conceived and designed the research, supervised project, edited manuscript, obtained funding.

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Correspondence to Kwan-Chang Kim, Shinkyu Choi or Suk Hyo Suh.

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Ethical approval

Experiments involving mice were approved by the Institutional Review Board for Human Research and Animal Care and Use Committee of the Ewha Womans University, Seoul, South Korea (EUM20-052). Experiments were conducted in accordance with the Declaration of Helsinki, Animal Care Guidelines of the Ewha Womans University Medical School, and Guide for the Care and Use of Laboratory Animals (NIH Publication no. 85 − 23, revised in 1996).

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Li, H., Kim, J.A., Jo, SE. et al. Modafinil exerts anti-inflammatory and anti-fibrotic effects by upregulating adenosine A2A and A2B receptors. Purinergic Signalling (2023). https://doi.org/10.1007/s11302-023-09973-8

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