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MicroRNA-376b is involved in the pathogenesis of thyroid-associated ophthalmopathy by regulating HAS2

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Abstract

Purpose

The aim of this study was to investigate the microRNA (miRNA) expression profile in peripheral blood mononuclear cells (PBMC) of thyroid-associated ophthalmopathy (TAO) patients and to explore the molecular mechanisms of MicroRNA-376b (miR-376b) in the pathogenesis of TAO.

Methods

PBMCs from TAO patients and healthy controls were analyzed by miRNA microarray to screen for the significantly differentially expressed miRNAs. The miR-376b expression in PBMCs were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). The downstream target of miR-376b was screened by online bioinformatics, and detected by qRT-PCR and Western blotting.

Results

Compared with normal controls, 26 miRNAs were significantly different in PBMCs of TAO patients (14 miRNAs were down-regulated and 12 miRNAs were up-regulated). Among them, miR-376b expression was significantly decreased in PBMCs from TAO patients compared to healthy controls. Spearman correlation analysis revealed that miR-376b expression in PBMCs was significantly negatively correlated with free triiodothyronine (FT3), and positively correlated with thyroid-stimulating hormone (TSH). MiR-376b expression was obviously reduced in 6T-CEM cells after triiodothyronine (T3) stimulation compared to controls. MiR-376b mimics significantly decreased hyaluronan synthase 2 (HAS2) protein expression and the mRNA expression of intercellular cell adhesion molecule-1 (ICAM1) and tumor necrosis factor-α (TNF-α) in 6T-CEM cells, whereas miR-376b inhibitors markedly elevated HAS2 protein expression and gene expression of ICAM1 and TNF-α.

Conclusions

MiR-376b expression in PBMCs was significantly decreased in PBMCs from TAO patients compared with the healthy controls. MiR-376b, regulated by T3, could modulate the expression of HAS2 and inflammatory factors. We speculate that miR-376b may be involved in the pathogenesis of TAO patients by regulating the expression of HAS2 and inflammatory factors.

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Acknowledgements

We are grateful to the nurses at the Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, for their help in blood collection.

Funding

This study was sponsored by the National Natural Science Foundation of China (Grant No. 81974105, 8227032471, 82200970), Medical Science and Technology Project of Xuhui District, Shanghai (SHXH201735).

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Author notes

  1. These authors contributed equally: Rongjiao Liu, Zhengqin Ye

Authors and Affiliations

  1. Department of Endocrinology and Metabolism, Tongji Hospital, School of Medicine, Tongji University, 200065, Shanghai, China

    Rongjiao Liu, Qi Liu, Miao Xuan, Ran Li, Liya Zhang, Keqin Zhang, Ping Fang & Ying Xue

  2. Department of Geriatrics, Shanghai Eighth People’s Hospital, 200235, Shanghai, China

    Rongjiao Liu

  3. Department of Geriatric Medicine, Tongji Hospital, School of Medicine, Tongji University, 200065, Shanghai, China

    Zhengqin Ye

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Contributions

Y.X. and P.F. contributed to the study conception and design. Material preparation was performed by Z.Y., R.L., L.Z., and K.Z. Data collection was performed by R.L., Q.L., and M.X. Analysis were performed by Z.Y. The first draft of the manuscript was written by R.L. and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Ping Fang or Ying Xue.

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Informed consent was obtained from all individual participants included in the study.

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This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Tongji Hospital, School of Medicine, Tongji University (Approval Number: K-2022-009).

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Liu, R., Ye, Z., Liu, Q. et al. MicroRNA-376b is involved in the pathogenesis of thyroid-associated ophthalmopathy by regulating HAS2. Endocrine 82, 87–95 (2023). https://doi.org/10.1007/s12020-023-03382-y

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