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Based on G-Series Mouse TH17 Array Study the Effect of Fluoride on C2C12 Cells Cytokines Expression

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Abstract

C2C12 cells were cultured on medium containing fluoride (0, 1, and 2.5 mmol/L) for 48 h to investigate the effect of excessive fluoride on T helper 17 (Th17)-related cytokine expression profile in skeletal muscle cells, and the culture supernatant was collected and subjected for the detection of 18 cytokines via Th17 array. Results showed that compared with the control group, no differential expression proteins (DEPs) were found in the 1 mmol/L fluoride group; however, eight DEPs were upregulated in the 2.5 mmol/L fluoride group, including macrophage inflammatory protein-3α (MIP-3α), interleukin-21 (IL-21), IL-13, IL-17F, IL-28A, transforming growth factor type beta 1 (TGF-β1), IL-23, and IL-17A. In addition, five DEPs (MIP-3α, IL-13, IL-21, TGF-β1, and IL-17F) were upregulated in the 2.5 mmol/L fluoride group compared with the 1 mmol/L fluoride group. Gene ontology analysis revealed that the positive regulation of cytokine production, cytokine activity, receptor ligand activity, and cytokine receptor binding accounted for high percent of DEPs present. Kyoto Encyclopedia of Genes and Genomes analysis showed that these DEPs primarily involved 12 pathways enriched in the cytokine–cytokine receptor interaction and IL-17 signaling pathway after 2.5 mmol/L fluoride treatment. The results indicated that fluoride might induce cytotoxicity by disturbing Th17-related cytokine expression.

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Acknowledgments

We thank Miss Chen Yu for her advice in cell culture.

Funding

This work is sponsored by the National Natural Science Foundation of China (Grant No. 31201963) and Natural Science Foundation of Henan (Grant No. 202300410120).

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Contributions

Shi-quan Zhu: Writing (original draft), software, validation, formal analysis, and data curation. Bian-hua Zhou: Ideas, formulation or evolution of overarching research goals and aims, methodology, review and editing, and project administration. Pan-pan Tan: Performing the experiments, formal analysis, and data curation. Jun Chai: Creation of cell culture model. Ya-ming Yu: Provision of study materials and reagents. Hong-wei Wang: Conceptualization, supervision, and funding acquisition.

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Correspondence to Bian-hua Zhou.

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Zhu, Sq., Zhou, Bh., Tan, Pp. et al. Based on G-Series Mouse TH17 Array Study the Effect of Fluoride on C2C12 Cells Cytokines Expression. Biol Trace Elem Res 199, 3402–3410 (2021). https://doi.org/10.1007/s12011-020-02464-6

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