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miR-7-5p Antagomir Protects Against Inflammation-Mediated Apoptosis and Lung Injury via Targeting Raf-1 In Vitro and In Vivo

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Abstract

Exacerbated inflammation and apoptosis are considered upstream events associated with acute lung injury (ALI). microRNAs are critical regulators of genes responsible for inflammation and apoptosis and are considered potential therapeutic targets for ameliorating ALI. This study was undertaken to uncover the role of miR-7-5p in LPS-induced lung injury. A LPS-induced inflammation model was established using BEAS-2B cells and C57BL/6 mice. Bioinformatics analysis and the luciferase reporter assay confirmed that Raf-1 is a target of miR-7-5p and that its expression was inversely correlated with expression of proinflammatory markers and miR-7-5p, whereas miR-7-5p inhibition in vitro led to subsequent restoration of Raf-1 expression and prevention of apoptosis. Intranasal (i.n.) administration of antagomir using the C57BL/6 mouse model further confirmed that miR-7-5p inhibition suppresses LPS-induced inflammation and apoptosis via modulating the miR-7-5p/Raf-1 axis. Our findings indicate that blocking miR-7-5p expression by antagomir protects mice from LPS-induced lung injury by suppressing inflammation and activation of mitochondria-mediated survival signalling. In conclusion, our findings demonstrate a previously unknown pathophysiological role of miR-7-5p in the progression of ALI, and targeted i.n. administration of miR-7-5p antagomir could aid in the development of potential therapeutic strategies against lung injury.

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AVAILABILITY OF DATA AND MATERIALS

All data generated or analyzed during this study are included in this article.

Abbreviations

COPD:

Chronic obstructive pulmonary disease

ERK:

Extracellular signal–regulated protein kinases

Bcl-2:

B-cell leukemia/lymphoma 2

Bax:

Bcl-2-associated X protein

MAPK:

Mitogen-activated protein kinase

UTR:

Untranslated region

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ACKNOWLEDGEMENTS

D.P. acknowledges the Department of Science and Technology (DST-India) for the WOS-A grant. D.P. is grateful to Dr. Rajesh Thimmaluappa, JSS Medical College, Mysuru, for providing BEAS-2B cells.

Funding

This work was supported by grants from the Department of Science and Technology (DST-India) (No. SR/ WOS-A/LS-50/2018).

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Authors and Affiliations

  1. Department of Biochemistry, CSIR-Central Food Technological Research Institute, Mysore, 570020, Karnataka, India

    Divya Peethambaran, Bijesh Puthusseri, Gyanendra Kumar, Rajasekar Janani, Parvatam Giridhar & Vallikannan Baskaran

  2. Department of Plant Cell Biotechnology, CSIR-Central Food Technological Research Institute, Mysore, 570020, Karnataka, India

    Bijesh Puthusseri, Gyanendra Kumar & Parvatam Giridhar

  3. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India

    Rajasekar Janani

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  1. Divya Peethambaran
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Contributions

Conceptualization, investigation and funding acquisition, methodology, writing original draft—D.P. Data curation, formal analysis and software—B.P. Validation, visualization, writing—review and editing—G.K. and R.J. Project administration, resources, supervision—V.B. and P.G.

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Correspondence to Vallikannan Baskaran.

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Ethics approval and all experiments were performed in accordance with the guidelines provided by Institute Animal Ethical Committee of CSIR–Central Food Technological Research Institute, Mysore, India (Approval No. CFT/IAEC/204/2020).

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Peethambaran, D., Puthusseri, B., Kumar, G. et al. miR-7-5p Antagomir Protects Against Inflammation-Mediated Apoptosis and Lung Injury via Targeting Raf-1 In Vitro and In Vivo. Inflammation 46, 941–962 (2023). https://doi.org/10.1007/s10753-023-01782-w

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