Abstract
In pulmonary fibrosis, the proliferation of fibroblasts and their differentiation into myofibroblasts is often caused by tissue damage, such as oxidative damage caused by reactive oxygen species, which leads to progressive rupture and thus destruction of the alveolar architecture, resulting in cell proliferation and tissue remodeling. Bezafibrate (BZF) is an important member of the peroxisome proliferator-activated receptor (PPARs) family agonists, used in clinical practice as antihyperlipidemic. However, the antifibrotic effects of BZF are still poorly studied. The objective of this study was to evaluate the effects of BZF on pulmonary oxidative damage in lung fibroblast cells. MRC-5 cells were treated with hydrogen peroxide (H2O2) to induce oxidative stress activation and BZF treatment was administered at the same moment as H2O2 induction. The outcomes evaluated were cell proliferation and cell viability; oxidative stress markers such as reactive oxygen species (ROS), catalase (CAT) levels and thiobarbituric acid reactive substances (TBARS); col-1 and α-SMA mRNA expression and cellular elasticity through Young's modulus analysis evaluated by atomic force microscopy (AFM). The H2O2-induced oxidative damage decreased the cell viability and increased ROS levels and decreased CAT activity in MRC-5 cells. The expression of α-SMA and the cell stiffness increased in response to H2O2 treatment. Treatment with BZF decreased the MRC-5 cell proliferation, ROS levels, reestablished CAT levels, decreased the mRNA expression of type I collagen protein (col-1) and α-smooth muscle actin (α-SMA), and cellular elasticity even with H2O2 induction. Our results suggest that BZF has a potential protective effect on H2O2-induced oxidative stress. These results are based on an in vitro experiment, derived from a fetal lung cell line and may emerge as a possible new therapy for the treatment of pulmonary fibrosis.
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When requested, the authors provided the data and materials used for the development of the manuscript.
Abbreviations
- α-SMA:
-
Alfa Smooth Muscle Actin and Type 1
- AFM:
-
Atomic Force Microscopy
- BZF:
-
Bezafibrate
- CAT:
-
Catalase
- Col-1:
-
Collagen 1
- DCFH-DA:
-
2’,7’-Dichlorodihydrofluorescein Diacetate
- DMEM:
-
Dulbecco´s Modified Eagle´s Medium
- ECM:
-
Extracellular Matrix
- FBS:
-
Fetal Bovine Serum
- H2O2 :
-
Hydrogen Peroxide
- PPARs:
-
Peroxisome Proliferator-Activated Receptor Agonists
- ROS:
-
Reactive Oxygen Species
- TBARS:
-
Thiobarbituric Acid-Reactive Substances
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Funding
This study was funded by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/Brazil; Financial Code 001), through a fellowship for the first author, and by National Council for Scientific and Technological Development (CNPq/Brazil) (Grant#311424/2018–0).
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DASM and JOR conceived and planned this project. CKR performed all experiments. BSB, BPC, GHV and ACS helped in cell viability and oxidative damage experiments. MSB, KGL, HBG, CL and MCRG performed the gene expression experiments. MVFD and KFR helped during the gene expression experiments as well as writing the manuscript. LSM, FMD, BSA and LLX performed the AFM experiments. CKR analyzed the data and wrote the manuscript with review and input from JOR, DASM, and MVFD. All authors approved the final manuscript. The authors declare that all data were generated in-house and that no paper mill was used.
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Reghelin, C.K., Bastos, M.S., de Souza Basso, B. et al. Bezafibrate reduces the damage, activation and mechanical properties of lung fibroblast cells induced by hydrogen peroxide. Naunyn-Schmiedeberg's Arch Pharmacol 396, 3857–3866 (2023). https://doi.org/10.1007/s00210-023-02595-2
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DOI: https://doi.org/10.1007/s00210-023-02595-2