类风湿关节炎患者CD4+ T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

doi:10.3969/j.issn.1674-8115.2024.10.006

上海交通大学学报（医学版） ›› 2024, Vol. 44 ›› Issue (10): 1249-1254.doi: 10.3969/j.issn.1674-8115.2024.10.006

• 论著 · 基础研究 • 上一篇

类风湿关节炎患者CD4⁺T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

李博¹(), 胡秋侠², 吴系美¹, 佘若男¹, 谭锦辉¹, 罗俊佳¹, 杨海涛¹, 张皓茹¹

^1.广东省深圳市龙华区人民医院风湿免疫科，深圳 518109
^2.广东省深圳市龙华区人民医院服务部，深圳 518109

收稿日期:2024-02-29 接受日期:2024-04-30 出版日期:2024-10-28 发布日期:2024-10-28
通讯作者: 李博 E-mail:friendlybo@163.com
基金资助:
广东省医学科研基金(A2013611);深圳市龙华区医疗卫生机构区级科研项目(2020023)

Expression of miR-146a in CD4⁺T lymphocytes of patients with rheumatoid arthritis and its correlation with inflammatory cytokines

LI Bo¹(), HU Qiuxia², WU Ximei¹, SHE Ruonan¹, TAN Jinhui¹, LUO Junjia¹, YANG Haitao¹, ZHANG Haoru¹

^1.Department of Rheumatology, The People's Hospital of Longhua Shenzhen, Guangdong Province, Shenzhen 518109, China
^2.Department of Outpatient Service, The People's Hospital of Longhua Shenzhen, Guangdong Province, Shenzhen 518109, China

Received:2024-02-29 Accepted:2024-04-30 Online:2024-10-28 Published:2024-10-28
Contact: LI Bo E-mail:friendlybo@163.com
Supported by:
Guangdong Medical Research Fund(A2013611);Scientific Research Projects of Medical and Health Institutions of Longhua District, Shenzhen(2020023)

摘要/Abstract

摘要：

目的·探讨类风湿关节炎（rheumatoid arthritis，RA）患者外周血CD4⁺T淋巴细胞中miR-146a的表达，及其与炎性细胞因子［肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）］的相关性。方法·选择2019年8月—2021年7月期间在深圳市龙华区人民医院就诊的活动期RA患者30例，并选择同期在该院体检的健康人群30例为健康对照者。分别抽取2组被试者的静脉血，先后分离获取外周血单个核细胞及CD4⁺ T淋巴细胞。采用实时荧光定量PCR（quantitative real-time PCR，qPCR）检测2组被试者外周血CD4⁺T淋巴细胞中miR-146a的表达，并采用酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）对其TNF-α、IL-6水平进行检测。使用miR-146a mimic转染RA患者的外周血CD4⁺T淋巴细胞，并于转染后再次对miR-146a、TNF-α及IL-6的表达进行检测。采用Pearson相关系数对转染前、后RA患者的miR-146a表达与TNF-α、IL-6表达的相关性进行分析。结果·转染miR-146a mimic前，与健康对照者相比，RA患者外周血CD4⁺T淋巴细胞中miR-146a、TNF-α及IL-6的表达较高（均P<0.001）。转染后RA患者外周血CD4⁺T淋巴细胞中miR-146a的表达水平较转染前有所升高、TNF-α及IL-6的表达则有所下降（均P<0.001）。Pearson相关分析的结果显示，转染前RA患者外周血CD4⁺ T淋巴细胞中miR-146a的表达水平与TNF-α、IL-6的表达水平呈正相关（r=0.959，P<0.001；r=0.916，P<0.001），转染后其miR-146a的表达水平与TNF-α、IL-6的表达水平亦呈正相关（r=0.971，P<0.001；r=0.861，P<0.001）。结论·miR-146a可能通过影响RA患者外周血CD4⁺T淋巴细胞分泌的TNF-α、IL-6等相关炎性细胞因子，在一定程度上参与RA的发病过程。

关键词: 类风湿关节炎, miR-146a, T淋巴细胞, 炎性细胞因子

Abstract:

Objective ·To investigate the expression of miR-146a in peripheral blood CD4⁺T lymphocytes of patients with rheumatoid arthritis (RA) and its correlation with inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Methods ·A total of 30 active RA patients who received medical treatment and 30 healthy controls who underwent physical examinations at the People's Hospital of Longhua, Shenzhen from August 2019 to July 2021 were selected. Peripheral blood mononuclear cells (PBMCs) and CD4⁺T lymphocytes were isolated from venous blood extracted from RA patients and healthy controls, respectively. Quantitative real-time PCR (qPCR) was used to detect the expression of miR-146a in peripheral blood CD4⁺ T lymphocytes, and enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of TNF-α and IL-6. After transfection of the peripheral blood CD4⁺T lymphocytes of RA patients with miR-146a mimic, the expression of miR-146a, TNF-α and IL-6 was detected again. The correlations between miR-146a expression and TNF-α and IL-6 expression in RA patients, both before and after transfection, were analyzed by using Pearson correlation coefficient. Results ·Before transfection with miR-146a mimic, the expression levels of miR-146a, TNF-α and IL-6 in peripheral blood CD4⁺T lymphocytes of RA patients were significantly higher than those of healthy controls (all P<0.001). After transfection, the expression of miR-146a in peripheral blood CD4⁺ T lymphocytes of RA patients was significantly higher, and the expression of TNF-α and IL-6 was significantly lower (all P<0.001). The results of Pearson correlation analysis showed that the expression of miR-146a in peripheral blood CD4⁺ T lymphocytes of RA patients, both before and after transfection, was positively correlated with the expression of TNF-α and IL-6, respectively (r=0.959, P<0.001; r=0.916, P<0.001; r=0.971, P<0.001; r=0.861, P<0.001). Conclusion ·miR-146a can regulate the levels of TNF-α and IL-6 in peripheral blood CD4⁺T lymphocytes of RA patients, indicating that miR-146a may play a role in the pathogenesis of RA.

Key words: rheumatoid arthritis (RA), miR-146a, T lymphocyte, inflammatory cytokine

中图分类号:

R593.22

李博, 胡秋侠, 吴系美, 佘若男, 谭锦辉, 罗俊佳, 杨海涛, 张皓茹. 类风湿关节炎患者CD4⁺T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性[J]. 上海交通大学学报（医学版）, 2024, 44(10): 1249-1254.

LI Bo, HU Qiuxia, WU Ximei, SHE Ruonan, TAN Jinhui, LUO Junjia, YANG Haitao, ZHANG Haoru. Expression of miR-146a in CD4⁺T lymphocytes of patients with rheumatoid arthritis and its correlation with inflammatory cytokines[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024, 44(10): 1249-1254.

图/表 4

表 1 qPCR引物序列

Tab 1 Primer sequences for qPCR

Gene	Forward primer (5' →3')	Reverse primer (5' →3')
miR-146a	GCAGGGTCCGAGGTATTCG	CGCGTGAGAACTGAATTCCAT
GAPDH	AAGGAAATGAATGGGCAGCC	TAGGAAAAGCATCACCCGGA

表 2 RA患者及健康对照者的人口信息学资料比较

Tab 2 Comparison of demographic data between RA patients and healthy controls

Item	RA patient (n=30)	Healthy control (n=30)	Z/χ² value	P value
Age/year	40.90 (26.00, 56.00)	43.30 (26.00, 56.25)	-0.564	0.573
Gender (male/female)/n	3/27	3/27	0.000	>0.999

表3 转染前，2组被试者外周血CD4+ T淋巴细胞中miR-146a、TNF-α及IL-6表达水平比较

Tab 3 Comparison of the expression levels of miR-146a， TNF-α and IL-6 in peripheral blood CD4+ T lymphocytes in RA patients and healthy controls before transfection

Index	RA patient (n=30)	Healthy control (n=30)	t value	P value
miR-146a	0.43±0.07	0.21±0.03	16.602	<0.001
TNF-α	0.60±0.08	0.25±0.03	23.567	<0.001
IL-6	1.76±0.09	0.89±0.03	52.808	<0.001

表4 miR-146a mimic转染前、后，RA患者外周血CD4+ T淋巴细胞中miR-146a、TNF-α及IL-6的表达水平比较

Tab 4 Comparison of the expression levels of miR-146a， TNF-α and IL-6 in peripheral blood CD4+ T lymphocytes before and after transfection in RA patients

Index	RA patient (n=30)		t value	P value
Index	Before transfection	After transfection	t value	P value
miR-146a	0.43±0.07	0.66±0.11	-9.840	<0.001
TNF-α	0.60±0.08	0.43±0.05	10.016	<0.001
IL-6	1.76±0.09	1.16±0.06	32.045	<0.001

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类风湿关节炎患者CD4⁺T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

Expression of miR-146a in CD4⁺T lymphocytes of patients with rheumatoid arthritis and its correlation with inflammatory cytokines

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类风湿关节炎患者CD4+ T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

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类风湿关节炎患者CD4⁺T淋巴细胞中miR-146a的表达及其与炎性细胞因子的相关性

Expression of miR-146a in CD4⁺T lymphocytes of patients with rheumatoid arthritis and its correlation with inflammatory cytokines