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格雷夫斯病中的环状RNA谱及hsa_circ_0090364的潜在功能

Circular RNA profile in Graves' disease and potential function of hsa_circ_0090364.

作者信息

Jiang Zhengrong, Huang Linghong, Chen Lijun, Zhou Jingxiong, Liang Bo, Bai Xuefeng, Wu Lizhen, Huang Huibin

机构信息

Department of Endocrinology, The Second affiliated Hospital of Fujian Medical University, Quanzhou, Fujian, China.

出版信息

Endocr Connect. 2022 Sep 28;11(11). doi: 10.1530/EC-22-0030. Print 2022 Nov 1.

Abstract

BACKGROUND

Graves' disease is a common autoimmune disease. Cytokines and their signalling pathways play a major part in the pathogenesis of Graves' disease; however, the underlying mechanism needs to be clarified.

AIMS

The aim of this study was to explore whether circular RNAs participate in the immunological pathology of Graves' disease via cytokine-related signalling pathways.

METHODS

Bioinformatics analysis was performed to identify differentially expressed circular RNAs and their targets and associated pathways. A total of three patients with Graves' disease and three sex- and age-matched healthy controls were enrolled for validation with microarray analysis and real-time quantitative PCR (qPCR). An additional 24 patients with Graves' disease and 24 gender- and age-matched controls were included for validation by real-time fluorescent qPCR. Flow cytometry and CCK8 assays were used to detect the apoptotic and proliferative levels of Jurkat cells (T lymphocytes) with the silenced expression of circRNA. ELISA was performed to detect the growth and apoptosis-related proteins. The competition mechanism of endogenous RNA was explored by real-time fluorescence qPCR.

RESULTS

A total of 366 significantly differentially expressed circular RNAs were identified in the Graves' disease group compared to healthy controls. The level of hsa_circ_0090364 was elevated in Graves' disease patients and positively correlated with thyroid-stimulating hormone receptor antibodies. Further analyses suggested that hsa_circ_0090364 may regulate the JAK-STAT pathway via the hsa-miR-378a-3p/IL-6ST/IL21R axis to promote cell growth.

CONCLUSIONS

These results provide novel clues into the pathophysiological mechanisms of Graves' disease and potential targets for drug treatment.

摘要

背景

格雷夫斯病是一种常见的自身免疫性疾病。细胞因子及其信号通路在格雷夫斯病的发病机制中起主要作用;然而,其潜在机制仍需阐明。

目的

本研究旨在探讨环状RNA是否通过细胞因子相关信号通路参与格雷夫斯病的免疫病理过程。

方法

进行生物信息学分析以鉴定差异表达的环状RNA及其靶标和相关通路。共纳入3例格雷夫斯病患者和3例年龄、性别匹配的健康对照,通过微阵列分析和实时定量聚合酶链反应(qPCR)进行验证。另外纳入24例格雷夫斯病患者和24例年龄、性别匹配的对照,通过实时荧光定量PCR进行验证。采用流式细胞术和CCK8法检测环状RNA沉默表达时Jurkat细胞(T淋巴细胞)的凋亡和增殖水平。采用酶联免疫吸附测定(ELISA)法检测生长和凋亡相关蛋白。通过实时荧光定量PCR探讨内源性RNA的竞争机制。

结果

与健康对照相比,格雷夫斯病组共鉴定出366种差异显著的环状RNA。格雷夫斯病患者中hsa_circ_0090364水平升高,且与促甲状腺激素受体抗体呈正相关。进一步分析表明,hsa_circ_0090364可能通过hsa-miR-378a-3p/IL-6ST/IL21R轴调控JAK-STAT通路以促进细胞生长。

结论

这些结果为格雷夫斯病的病理生理机制提供了新线索,并为药物治疗提供了潜在靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b0fb/9578071/5456e3dec15c/EC-22-0030fig1.jpg

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