Inhibition of MicroRNA-326 in a NOD.H-2 Mouse Model of Autoimmune Thyroiditis

Overview

Journal Front Immunol

Specialty Allergy & Immunology

Date 2021 Jun 18

PMID 34140947

Citations 3

Authors

Na Zhao

Zhenzhen Wang

Xuejiao Cui

Shuo Wang

Chenling Fan

Yushu Li

Zhongyan Shan

Weiping Teng

Affiliations

Soon will be listed here.

Abstract

Background: Previous studies reported that various miRNAs participate in autoimmune diseases, but the potential regulatory mechanism of miRNAs in autoimmune thyroiditis (AIT) needs further exploration.

Objective: This study aimed to further verify that miR-326 contributes to AIT by regulating Th17/Treg balance through Ets-1 using lentiviral gene delivery through tail vein and thyroid injection in NOD.H-2 mice.

Materials And Methods: Five-week-old NOD.H-2 mice were divided randomly into tail vein and thyroid injection groups, and each received either mmu-miR-326 sponge (LV-sponge) or lentiviral vector control. Mice were divided for tail vein injection: the therapeutic LV-ctrl, therapeutic LV-sponge, prophylactic LV-ctrl, and prophylactic LV-sponge groups. The control group was fed high-iodine water without vein injection. The thyroid infiltration of lymphocytes and serum TgAb value were investigated by thyroid hematoxylin and eosin (HE) staining and ELISA, respectively. Ets-1 and lymphocyte counts were measured by RT-PCR, western blotting, and flow cytometry. The thyroid CD4IL-17a cells and CD4Ets-1 cells were detected by immunofluorescence, and the serum cytokines were tested by ELISA.

Results: In the tail vein injection groups, the thyroid inflammatory score and serum TgAb titer were significantly lower in the LV-sponge groups than in the control and LV-ctrl groups while Ets-1 protein expression in mouse spleens was increased in the LV-sponge groups. Moreover, Th17/Treg ratio declined in the LV-sponge group and decreased significantly in the prophylactic LV-sponge group ( = 0.036) tested by flow cytometry. Immunofluorescence showed that, in LV-sponge groups, CD4IL-17a cells were decreased significantly ( = 0.001), while CD4Ets-1 cells were increased significantly in the LV-sponge group ( = 0.029). The serum IL-17/IL-10 was decreased significantly in the LV-sponge group ( < 0.05). In the thyroid injection groups, the thyroid inflammatory score and serum TgAb titer in the LV-sponge group decreased significantly compared with those in the LV-ctrl group ( < 0.05). In addition, in LV-sponge groups, CD4IL-17a cells were decreased, while CD4Ets-1 cells were increased significantly in the inhibition group evaluated by immunofluorescence. Moreover, tail vein injection of LV-sponge resulted in much lower TgAb levels in thyroiditis compared with thyroid injection.

Conclusion: MiR-326 targeted therapy may be a promising approach for AIT. In addition, tail vein injection may achieve a better intervention effect than thyroid injection.

Citing Articles

miR-142-3p encapsulated in T lymphocyte-derived tissue small extracellular vesicles induces Treg function defect and thyrocyte destruction in Hashimoto's thyroiditis.

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MiR-326 regulates cell proliferation and apoptosis in fibroblast-like synoviocytes in rheumatoid arthritis.

Yiou W, Zhihong W, Shibai Z, Shanni L, Wenwei Q Hum Cell. 2023; 36(3):987-996.

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Leptin receptor antagonist attenuates experimental autoimmune thyroiditis in mice by regulating Treg/Th17 cell differentiation.

Wang W, Zhang B, Jiang Q, Zhao H, Xu Q, Zeng Y Front Endocrinol (Lausanne). 2022; 13:1042511.

PMID: 36339447 PMC: 9630560. DOI: 10.3389/fendo.2022.1042511.

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