miR-155通过SOCS1/STAT3途径调控类风湿性关节炎中炎症反应和Th17/Treg失衡

doi:10.3969/j.issn.1006-5725.2024.13.005

摘要/Abstract

摘要：

目的在类风湿性关节炎（RA）中探究miR-155和细胞因子信号转导抑制因子1（SOCS1）的表达及作用机制。方法采用RT-PCR和流式细胞术检测miR-155和Th17、Treg细胞在RA患者（RA组）和对照组（HC组）外周血中的表达差异；生物信息学和双荧光素酶基因报告实验检测miR-155和SOSC1的调控关系；分离RA患者外周CD4⁺T细胞，将沉默miR-155与SOCS1表达的miR-155 inhibitor和si-SOCS1及各自的阴性对照序列分别或联合转染入CD4⁺T细胞中，并将细胞分为：miR-NC组、miR-155 inhibitor组、miR-155 inhibitor+si-NC组和miR-155 inhibitor+si-SOCS1组。使用Th17诱导分化液处理上述细胞后，采用流式细胞术检测各组CD4⁺T细胞中Th17比率，Western blot实验检测细胞中p-STAT3/STAT3比值。结果与HC组相比，RA患者中miR-155、Th17比率升高（P < 0.01），Treg细胞比率降低（P < 0.01）；miR-155可靶向抑制SOCS1表达。与miR-NC组相比，miR-155 inhibitor组、miR-155 inhibitor+si-NC组和miR-155 inhibitor+si-SOCS1组中Th17比率、p-STAT3/STAT3比值均降低（P < 0.01）；与miR-155 inhibitor组相比，miR-155 inhibitor+si-SOCS1组CD4⁺T细胞中Th17比率、p-STAT3/STAT3比值均升高（P < 0.05）。结论在RA患者中表达升高的miR-155可能通过SOCS1/STAT3途径来介导CD4⁺T细胞的Th17分化，从而参与RA患者外周Th17/Treg细胞失衡。

关键词: 类风湿性关节炎, miR-155, 细胞因子信号转导抑制因子1, Th17/Treg失衡

Abstract:

Objective This research aimed to investigate the expression and mechanism of miR-155 and suppressor of cytokine signaling 1 （SOCS1） in rheumatoid arthritis （RA）. Methods RT-PCR and flow cytometry were applied to detect the expression differences of miR-155， Th17， and Treg cells in peripheral blood of RA patients （RA group） and control group （HC group）. Bioinformatics analysis and dual-luciferase reporter assay were conducted to investigate the regulatory relationship between miR-155 and SOCS1. CD4⁺T cells were isolated from peripheral blood of RA patients and transfected with miR-155 inhibitor， si-SOCS1， and their respective negative control sequences， and divided into four groups： miR-NC group， miR-155 inhibitor group， miR-155 inhibitor+si-NC group， and miR-155 inhibitor+si-SOCS1 group. The cells were treated with Th17-inducing differentiation medium， and flow cytometry was used to determine the ratio of Th17 cells in each group of CD4⁺T cells. Western blot was used to determine the ratio of p-STAT3/STAT3 in the cells. Results Compared to the HC group， RA patients showed increased expression of miR-155 and Th17 ratio （P < 0.01）， and decreased Treg cell ratio （P < 0.01）. MiR-155 could target and inhibit the expression of SOCS1. Compared to the miR-NC group， the miR-155 inhibitor group， miR-155 inhibitor+si-NC group， and miR-155 inhibitor+si-SOCS1 group showed decreased Th17 ratio and p-STAT3/STAT3 ratio （P < 0.01）. Compared to the miR-155 inhibitor group， the miR-155 inhibitor+si-SOCS1 group exhibited increased Th17 ratio and p-STAT3/STAT3 ratio in CD4⁺T cells （P < 0.05）. Conclusion Elevated expression of miR-155 in RA patients may mediate the differentiation of CD4⁺ T cells into Th17 cells through the SOCS1/STAT3 pathway， contributing to the imbalance of Th17/Treg cells in peripheral blood of RA patients.

Key words: rheumatoid arthritis, miR-155, suppress of cytokine signaling 1 （SOCS1）, Th17/Treg imbalance

中图分类号:

R539.22

张玉红,单新洁,周俊. miR-155通过SOCS1/STAT3途径调控类风湿性关节炎中炎症反应和Th17/Treg失衡[J]. 实用医学杂志, 2024, 40(13): 1791-1796.

Yuhong ZHAN,Xinjie SHAN,Jun. ZHOU. MiR⁃155 regulates inflammatory responses and Th17/Treg imbalances in rheumatoid arthritis through the SOCS1/STAT3 pathway[J]. The Journal of Practical Medicine, 2024, 40(13): 1791-1796.

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临床特征	RA组（n = 20）	HC组（n = 15）	t/χ²值	P值
性别（男/女，例）	11/9	8/7	3.18	0.26
年龄（岁）	45.81 ± 11.54	44.62 ± 13.17	1.49	0.17
患病时间（年）	10.05 ± 4.28	-	9.06	< 0.01
DAS28（分）	3.42 ± 1.03	-	12.81	< 0.01
ESR（mm/h）	20.40 ± 14.51	-	5.42	< 0.01
CRP（mg/L）	5.29 ± 2.37	-	8.61	< 0.01
RF（IU/mL）	31.55 ± 8.49	-	14.34	< 0.01
Anti-CCP（IU/mL）	36.13 ± 11.76	-	11.85	< 0.01
TJC（个）	3.02 ± 2.12	-	5.49	< 0.01
SJC（个）	2.73 ± 2.04	-	5.16	< 0.01

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