Effect of N-acetyltransferase 10 on bleomycin-induced oxidative stress in lung epithelial cells

doi:10.3969/j.issn.1006-7795.2026.01.015

Abstract

Abstract: Objective To investigate the role of N-acetyltransferase 10 (NAT10) in pulmonary fibrosis (PF) and its association with oxidative stress. Methods Public transcriptome datasets (GEO: GSE110147, GSE282477, GSE128033) were integrated to analyze NAT10 mRNA levels in pulmonary fibrosis (PF). A murine PF model was established by intratracheal Bleomycin (BLM) administration. The expression of NAT10 protein in lung tissue was assessed by immunofluorescence. Human bronchial epithelial cell line BEAS-2B was stimulated by BLM to establish an in vitro fibrosis model. The mRNA expression of fibrotic markers and NAT10 were analyzed by using reverse transcription polymerase chain reaction (RT-qPCR), while the protein levels of inflammatory factors were evaluated by enzyme-linked immunosorbent assay (ELISA). To study the function of NAT10, BEAS-2B cells with NAT10 inhibition were treated with BLM. Intracellular reactive oxygen species (ROS) levels were analyzed by flow cytometry. The concentrations of interleukin 6 (IL-6), transforming growth factor-β1 (TGF-β1), superoxide dismutase (SOD) activity, and the lipid peroxidation product malondialdehyde (MDA) content were measured via ELISA. Results Based on public transcriptome analysis, NAT10 mRNA expression was elevated to 1.05- and 1.38-fold of controls in lung tissues from idiopathic pulmonary fibrosis (IPF) patients (GSE110147) and BLM-induced fibrotic mice (GSE282477), respectively (both P<0.05). Single-cell RNA sequencing of IPF lungs (GSE128033) indicated that bronchiolar club and other epithelial cells exhibited the most pronounced NAT10 upregulation compared to other pulmonary cell types (all P<0.05). Immunofluorescence quantification confirmed a 1.94-fold increase in NAT10 protein in BLM-induced fibrotic murine lungs versus model controls (P<0.05). In BEAS-2B cells, BLM stimulation for 48 hours induced fibrotic changes and increased NAT10 mRNA to 1.36-fold of baseline (P<0.05). This was accompanied by elevated supernatant levels of ROS, MDA content, TGF-β1 and IL-6 levels, alongside reduced SOD activity (all P<0.05). NAT10 knockdown rescued these alterations, increasing SOD activity and decreasing ROS levels, MDA content, TGF-β1, and IL-6 levels compared to the BLM-stimulated group (all P<0.05). Conclusion NAT10 is a critical regulator of oxidative stress injury in pulmonary fibrosis. Its expression is significantly elevated in fibrotic tissues and cellular models, suggesting a possible correlation with disease severity. At the cellular level, targeting NAT10 effectively mitigates BLM-induced oxidative stress and fibrotic responses. Further animal studies will help clarify its therapeutic potential.

Key words: N-acetyltransferase 10, pulmonary fibrosis, oxidative stress, BEAS-2B cells, bleomycin, gene knockdown, reactive oxygen species

CLC Number:

R563
R392.5

Wang Yanran, Xu Yuling, Song Nan. Effect of N-acetyltransferase 10 on bleomycin-induced oxidative stress in lung epithelial cells[J]. Journal of Capital Medical University, 2026, 47(1): 115-125.

References

［1］Chapman H A. Disorders of lung matrix remodeling［J］. J Clin Invest, 2004, 113(2): 148－157.
［2］Phan T H G, Paliogiannis P, Nasrallah G K, et al. Emerging cellular and molecular determinants of idiopathic pulmonary fibrosis［J］. Cell Mol Life Sci, 2021, 78(5): 2031－2057.
［3］Chambers R C, Mercer P F. Mechanisms of alveolar epithelial injury, repair, and fibrosis［J］. Ann Am Thorac Soc, 2015, 12(Suppl 1): S16-S20.
［4］Song L C, Li K, Chen H Y, et al. Cell cross-talk in alveolar microenvironment: from lung injury to fibrosis［J］. Am J Respir Cell Mol Biol, 2024, 71(1): 30－42.
［5］Henderson N C, Rieder F, Wynn T A. Fibrosis: from mechanisms to medicines［J］. Nature, 2020, 587(7835): 555－566.
［6］Pott H, Sykes D L, Charriot J, et al. Breathing barriers: bridging lung health, research, and awareness［J］. Lancet Respir Med, 2025, 13(8): 665－667.
［7］Spagnolo P, Guler S A, Chaudhuri N, et al. Global epidemiology and burden of interstitial lung disease［J］. Lancet Respir Med, 2025, 13(8): 739－755.
［8］Liu T J, De Los Santos F G, Phan S H. The bleomycin model of pulmonary fibrosis［J］. Methods Mol Biol, 2017, 1627: 27－42.
［9］El-Baz L M, Shoukry N M, Hafez H S, et al. Experimental mouse model of bleomycin-induced pulmonary fibrosis［J］. Int J Cancer Biomed Res, 2020, 4(3): 1－8.
［10］Burger R M, Peisach J, Horwitz S B. Activated bleomycin. A transient complex of drug, iron, and oxygen that degrades DNA［J］. J Biol Chem, 1981, 256(22): 11636－11644.
［11］Ayilya B L, Balde A, Ramya M, et al. Insights on the mechanism of bleomycin to induce lung injury and associated in vivo models: a review［J］. Int Immunopharmacol, 2023, 121: 110493.
［12］Teixeira K C, Soares F S, Rocha L G C, et al. Attenuation of bleomycin-induced lung injury and oxidative stress by N-acetylcysteine plus deferoxamine［J］. Pulm Pharmacol Ther, 2008, 21(2): 309－316.
［13］Liu X J, Chen Z H. The pathophysiological role of mitochondrial oxidative stress in lung diseases［J］. J Transl Med, 2017, 15(1): 207.
［14］Rogers L K, Cismowski M J. Oxidative stress in the lung-the essential paradox［J］. Curr Opin Toxicol, 2018, 7: 37－43.
［15］Della Latta V, Cecchettini A, Del Ry S, et al. Bleomycin in the setting of lung fibrosis induction: from biological mechanisms to counteractions［J］. Pharmacol Res, 2015, 97: 122－130.
［16］Cheresh P, Kim S J, Tulasiram S, et al. Oxidative stress and pulmonary fibrosis［J］. Biochim Biophys Acta, 2013, 1832(7): 1028－1040.
［17］Liu J T, Han X Y, Zhang T Y, et al. Reactive oxygen species (ROS) scavenging biomaterials for anti-inflammatory diseases: from mechanism to therapy［J］. J Hematol Oncol, 2023, 16(1): 116.
［18］Wang Z W, Chen S Y, Guo W X, et al. Oxidative stress promotes intervertebral disc degeneration through NAT10-mediated IKKβ N4-acetylcytidine modification［J］. Cell Signal, 2025, 134: 111918.
［19］Qu Z Z, Pang X C, Mei Z T, et al. The positive feedback loop of the NAT10/Mybbp1a/p53 axis promotes cardiomyocyte ferroptosis to exacerbate cardiac I/R injury［J］. Redox Biol, 2024, 72: 103145.
［20］Wu S S, Yin L J, Han K, et al. NAT10 accelerates pulmonary fibrosis through N4-acetylated TGFB1-initiated epithelial-to-mesenchymal transition upon ambient fine particulate matter exposure［J］. Environ Pollut, 2023, 322: 121149.
［21］Spagnolo P, Kropski J A, Jones M G, et al. Idiopathic pulmonary fibrosis: disease mechanisms and drug development［J］. Pharmacol Ther, 2021, 222:107798.
［22］Otoupalova E, Smith S, Cheng G J, et al. Oxidative stress in pulmonary fibrosis［J］. Compr Physiol, 2020, 10(2): 509－547.
［23］Veith C, Boots A W, Idris M, et al. Redox imbalance in idiopathic pulmonary fibrosis: a role for oxidant cross-talk between NADPH oxidase enzymes and mitochondria［J］. Antioxid Redox Signal, 2019, 31(14): 1092－1115.
［24］Ranneh Y, Ali F, Akim A M, et al. Crosstalk between reactive oxygen species and pro-inflammatory markers in developing various chronic diseases: a review［J］. Appl Biol Chem, 2017, 60(3): 327－338.
［25］Richter K, Konzack A, Pihlajaniemi T, et al. Redox-fibrosis: impact of TGFβ1 on ROS generators, mediators and functional consequences［J］. Redox Biol, 2015, 6: 344－352.
［26］Kinnula V L, Fattman C L, Tan R J, et al. Oxidative stress in pulmonary fibrosis: a possible role for redox modulatory therapy［J］. Am J Respir Crit Care Med, 2005, 172(4): 417－422.
［27］Makena P, Kikalova T, Prasad G L, et al. Oxidative stress and lung fibrosis: towards an adverse outcome pathway［J］. Int J Mol Sci, 2023, 24(15): 12490.

[1]	Cui Wei, Pang Qidan, Xiang Hanyue, Xiao Nao, Jiang Dechun, Li Shen, Shen Guangli. Detection of cerebral ischemia-reperfusion injury using BIBP-H fluorescent probe [J]. Journal of Capital Medical University, 2025, 46(1): 76-82.
[2]	Li Yakun, Li Wei, Yang Xueqi, Shi Wenjuan, Zhu Yuequan, Zhao Yongmei. Effect of mitochondrial reactive oxygen species inhibitor EUK-134 on mitochondrial associated proteins PGC-1α and VDAC1 in rats with cerebral ischemia-reperfusion injury [J]. Journal of Capital Medical University, 2024, 45(6): 1016-1022.
[3]	Jiang Yumei, Zhang Ziwei, Wang Jitian, Yang Yu. Effect of nitric oxide donor drugs on the antibacterial activity of Porphyromonas gingivalis in vitro [J]. Journal of Capital Medical University, 2024, 45(1): 104-110.