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Intermittent hypoxia induces myofibroblast differentiation and extracellular matrix production of MRC5s via HIF-1α-TGF-β/Smad pathway

  • Sleep Breathing Physiology and Disorders • Original Article
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Abstract

Purpose

To investigate whether or not intermittent hypoxia (IH), the main characteristic of obstructive sleep apnea (OSA) may affect the myofibroblast differentiation and extracellular matrix (ECM) production of lung fibroblast through the HIF-1α-TGF-β/Smad pathway and assess the interventional role of a HIF-1α inhibitor, 2-methoxyestradiol (2-ME2).

Method

The human lung fibroblast MRC5 cells were exposed to normoxia or IH conditions, and the expression of myofibroblast differentiation marker α-smooth muscle actin (α-SMA) and ECM protein collagen I were evaluated. To clarify the underlying mechanism, the expression level of HIF-1α, TGF-β, and p-Smads/Smads were measured and the effects of inhibiting HIF-1α with 2-ME2 on the α-SMA expression level and ECM production through the TGF-β/Smad pathway were assessed. Si HIF-1α was applied to genetically inhibit HIF-1α in MRC5 cells, and the related proteins were assessed.

Results

IH increased the protein and mRNA expression of Collagen I and α-SMA of MRC5 cells in a time-dependent manner. IH activated the protein and mRNA level of HIF-1α and TGF-β and increased the phosphorylation of Smad2/Smad3 of MRC5 cells in a time-dependent manner. 2-ME2 inhibited the activation of HIF-1α induced by IH and decreased overexpression of TGF-β, p-Smad2/Smad2, and p-Smad3/Smad3, which in turn partially reversed the upregulation of α-SMA and Collagen I induced by IH in MRC5 cells. When HIF-1α was successfully silenced by si-HIF-1α, upregulation of TGF-β induced by intermittent hypoxia was partially decreased.

Conclusions

This study showed that IH contributes to myofibroblast differentiation and excessive ECM production of MRC5 cells through activation of the HIF-1α-TGF-β/Smad pathway. 2-ME2 partially attenuated myofibroblast differentiation induced by IH by inhibiting the HIF-1α-TGF-β/Smad pathway.

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Data availability

Data available on request from the authors.

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Funding

Funding for this study included grants from the Chinese National Natural Science Foundation (Nos. 81970084 and 81670084), Chinese National Key Research and Development Program (No. 2016YFC1304502), and the Tianjin Key Research and Development Program (No. 20YFZCSY00390).

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Author notes

  1. Maoli Liang and Liang Si contributed equally to this work.

Authors and Affiliations

  1. Department of Respiratory and Critical Care Medicine, Tianjin Medical University General Hospital, Tianjin, People’s Republic of China

    Maoli Liang, Liang Si, Zhi Yu, Hui Ding, Le Wang, Xing Chen, Baoyuan Chen, Jing Zhang & Jie Cao

  2. Department of Respiratory and Critical Care Medicine, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, People’s Republic of China

    Maoli Liang

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  1. Maoli Liang
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  2. Liang Si
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  5. Le Wang
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Correspondence to Jing Zhang or Jie Cao.

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Liang, M., Si, L., Yu, Z. et al. Intermittent hypoxia induces myofibroblast differentiation and extracellular matrix production of MRC5s via HIF-1α-TGF-β/Smad pathway. Sleep Breath 28, 291–300 (2024). https://doi.org/10.1007/s11325-023-02889-y

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  • DOI: https://doi.org/10.1007/s11325-023-02889-y

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