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Calcitonin Gene-Related Peptide Attenuates Hyperoxia-Induced Oxidative Damage in Alveolar Epithelial Type II Cells Through Regulating Viability and Transdifferentiation

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Abstract

As a stem cell of alveolar epithelium, the physiological status of alveolar epithelium type II cells (AECII) after hyperoxia exposure is closely related to the occurrence of hyperoxia-induced lung injury and the restoration of normal morphological function of damaged alveolar epithelium. However, the relevant mechanisms involved are not very clear. Therefore, this study aimed to explore the effect of calcitonin gene-related peptide (CGRP) on AECII exposed to hyperoxia and its potential mechanisms. The AECII viability was detected using MTT assay. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were detected by spectrophotometry. The transdifferentiation capacity of AECII was evaluated by flow cytometry. The expression levels of Notch1, Hes, HERP, and AECII markers were detected using immunohistochemistry and/or RT-qPCR or immunofluorescence. ELISA was used for the determination of inflammatory markers. The results showed that CGRP significantly promoted cell viability, and markedly suppressed hyperoxia-induced transdifferentiation of AECII; these biological alterations were coincided with decreased MDA level, increased SOD activity, and activated Notch signaling pathway (upregulated expression levels of Notch1, Hes, and HERP). Notably, the in vitro effects of CGRP on Notch signaling pathway were further investigated in animal model, and the HE staining results showed that CGRP reduced in vivo oxidative injury and inflammation in hyperoxia-treated AECII through the promotion of structural and functional regeneration, accompanied by elevated Notch1 expression and activated Notch signaling cascade as shown by immunohistochemistry and QPCR, respectively. Immunohistochemistry of APQ-5 and SPC indicated that CGRP reversed the transdifferentiation of AECIIs in vivo. Our current results were consistent across both in vitro and in vivo settings, and provide a new direction for the prevention and treatment of bronchopulmonary dysplasia (BPD).

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Funding

This work was supported by the Natural Science Foundation of Shenzhen Science and Technology Innovation Commission in 2020 [grant number JCYJ20190813141207091].

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  1. Neonatal Intensive Care Unit, Women and Children Health Institute of Futian, University of South China, Futian District, 2002 Jintian South Road, ShenzhenGuangdong Province, 518045, No, China

    Jian Deng, Shao-Hua Wang, Xue-Mei Zheng & Zan-Mei Tang

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  1. Jian Deng
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  2. Shao-Hua Wang
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JD wrote the manuscript, obtained grants from the national funding agency, and designed the work. SW designed the work and supervised the study to be implemented smoothly. XZ and ZT contributed to the analysis and interpretation of data.

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Correspondence to Shao-Hua Wang.

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The study protocol was approved by the Animal Care and Use Committee of University of South China. All animal experiments were performed with the minimum number of animals, and care was taken to minimize any suffering to the animals with the recommendations of the NIH Guide for the Care and Use of Laboratory Animals.

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Deng, J., Wang, SH., Zheng, XM. et al. Calcitonin Gene-Related Peptide Attenuates Hyperoxia-Induced Oxidative Damage in Alveolar Epithelial Type II Cells Through Regulating Viability and Transdifferentiation. Inflammation 45, 863–875 (2022). https://doi.org/10.1007/s10753-021-01591-z

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