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Notch signaling regulates Th17 cells differentiation through PI3K/AKT/mTORC1 pathway and involves in the thyroid injury of autoimmune thyroiditis

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Abstract

Purpose

Autoimmune Thyroiditis (AIT) is the most common thyroid disease; however, there were no measures to prevent the progression of the disease. The present study attempts to identify that Notch signaling regulates the differentiation of T helper 17 (Th17) cells by activating downstream Phosphatidylinositol-3 kinase/protein kinase/mechanistic target of rapamycin complex 1 (PI3K/AKT/mTORC1) pathway participating in the thyroid injury of the experimental autoimmune thyroiditis (EAT).

Methods

In vivo experiments, mice were randomly divided into 4 groups: a control group, an EAT group, and two groups with LY294002 treatment (pTg plus 25 mg/kg or 50 mg/kg LY294002, respectively). The degrees of thyroiditis were evaluated, and the percentage of Th17 cells, expression of interleukin-17A (IL-17A), and the main components of the Notch–PI3K signaling pathway were detected in different groups. In vitro experiments, two different dosages of LY294002 (25 and 50 μM) were used to intervene splenic mononuclear cells (SMCs) from EAT mice to further evaluate the regulatory effect of Notch–PI3K pathway on Th17 cells.

Results

Our data demonstrate that the infiltration of Th17 cells and the expressions of IL-17A, Notch, hairy and split 1 (Hes1), p‑AKT (Ser473), p‑AKT (Thr308), p‑mTOR (Ser2448), S6K1, and S6K2 increased remarkably in EAT mice. After PI3K pathway was blocked, the degrees of thyroiditis were significantly alleviated, and the proportion of Th17 cells, the expression of IL-17A, and the above Notch–PI3K pathway-related molecules decreased in a dose-dependent manner. Additionally, the proportion of Th17 cells was positively correlated with the concentration of serum thyroglobulin antibody (TgAb), IL-17A, and Notch–PI3K pathway-related molecules mRNA levels.

Conclusions

Notch signal promotes the secretion of IL-17A from Th17 cells by regulating the downstream PI3K/AKT/mTORC1 pathway through Hes-Phosphatase and tensin homolog (PTEN) and participates in thyroid autoimmune damage, and the PI3K pathway inhibitor may play important effects on AIT by affecting Th17 cells differentiation.

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Data availability

The data used to support the findings of this study are available from the corresponding authors upon request.

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Acknowledgements

The authors are gratefully for the contributions from Dr. Chen Li and Dr. Lijuan Yang from Medical Research Center of Binzhou Medical University Hospital for their assistance of experimental technology during all experiments.

Funding

This work was supported by grants from the Natural Science Foundation of Shandong Province (No. ZR2013HM049, ZR2022MH175), Key Technology Research and Development Program of Shandong (No. 2016GSF201021), Project of Shandong Province Higher Educational Science and Technology program (No. J16LL01), Reserve Leading Talents Project of Binzhou Medical University Hospital (No. JC2019-03), Scientific Research and Innovation Team Project of Binzhou Medical University Hospital (No. 202031), and Innovation Program of Post-graduate Education of Shandong Province (No. 20038612).

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Authors and Affiliations

  1. Department of Endocrinology and Metabolism, Binzhou Medical University Hospital, Binzhou, 256600, People’s Republic of China

    C. He, Y. Li, L. Gan & H. Xue

  2. Department of Dermatology, Binzhou Medical University Hospital, Binzhou, 256600, People’s Republic of China

    Y. Lin & L. Ma

  3. Nanchang University Queen Mary School, Nanchang, 330031, People’s Republic of China

    B. Zhang

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Contributions

HX and LM were responsible for the design of this study, and reviewed and edited this article. CH, YL, LG, YL, and BZ performed the experiments. CH participated in data analysis and wrote the first draft of the manuscript, especially for the development of the experimental models and the in vivo and in vitro experiments.

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Correspondence to H. Xue.

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Human participants were not involved in the present study, and the animal study was reviewed and approved (Approval No. 20220128-73) by the Laboratory Animal Ethics Committee of Binzhou Medical University Hospital (Binzhou, China).

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He, C., Li, Y., Gan, L. et al. Notch signaling regulates Th17 cells differentiation through PI3K/AKT/mTORC1 pathway and involves in the thyroid injury of autoimmune thyroiditis. J Endocrinol Invest (2024). https://doi.org/10.1007/s40618-023-02293-z

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