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

doi:10.3969/j.issn.1674-8115.2024.10.006

Abstract

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

CLC Number:

R593.22

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.

Figures/Tables 4

TrendMD

References 20

1	LIU M L, REN T L, LIN Z, et al. Upregulated miR-146a expression in peripheral blood relates to Th17 and Treg imbalance in elder rheumatoid arthritis patients[J]. Lifestyle Genom, 2022, 15(3): 98-106.
2	雷海桃, 田雪梅, 金芳全. 细胞因子信号转导抑制因子与类风湿关节炎的相关性研究进展[J]. 上海交通大学学报(医学版), 2022, 42(7): 945-951.
	LEI H T, TIAN X M, JIN F Q. Advances in the correlation between cytokine signal transduction inhibitors and rheumatoid arthritis[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(7): 945-951.
3	ISLAM Z U, BANEEN U, KHALIQ T, et al. Association analysis of miRNA-146a and miRNA-499 polymorphisms with rheumatoid arthritis: a case-control and trio-family study[J]. Clin Exp Med, 2023, 23(5): 1667-1675.
4	ALI Y, KHAN A, AKHTAR M, et al. Exploring the associations and molecular impacts of miR-146a/rs2910164 and miR-196a2/rs185070757 with rheumatoid arthritis in a Pakistani population[J]. Lifestyle Genom, 2023, 16(1): 139-150.
5	ZHANG L L, WU X X, WANG X F, et al. Genetic variant in microRNA-146a gene is associated with risk of rheumatoid arthritis[J]. Ann Med, 2021, 53(1): 824-829.
6	李博. miR-146a在类风湿关节炎发病机制中的作用及其与中医体质的相关性研究[D]. 广州: 南方医科大学, 2020.
	LI B. miR-146a in the pathogenesis of rheumatoid arthritis and its correlation with traditional Chinese medical constitutions[D]. Guangzhou: Southern Medical University, 2020.
7	李博, 胡秋侠, 谭锦辉, 等. 类风湿关节炎患者中医体质分布及miR-146a表达[J]. 中国中西医结合杂志, 2019, 39(9): 1045-1049.
	LI B, HU Q X, TAN J H, et al. Chinese medical constitution and expression of miR-146a in rheumatoid arthritis patients[J]. Chinese Journal of Integrated Traditional and Western Medicine, 2019, 39(9): 1045-1049.
8	胡秋侠, 李博, 佘若男, 等. miR-146a rs2910164位点多态性与类风湿关节炎病情活动的相关性[J]. 中华医学遗传学杂志, 2019, 36(5): 505-507.
	HU Q X, LI B, SHE R N, et al. Association of polymorphisms of miR-146a rs2910164 locus with clinical features of rheumatoid arthritis[J]. Chinese Journal of Medical Genetics, 2019, 36(5): 505-507.
9	李博, 胡秋侠, 谭锦辉, 等. 类风湿关节炎中miR-146a多态性与临床表现及血清中肿瘤坏死因子α水平之间的相关性[J]. 中国现代应用药学, 2019, 36(4): 471-474.
	LI B, HU Q X, TAN J H, et al. miR-146a rs2910164 polymorphism correlates with extraarticular manifestations and the serum TNF-α level in patients with rheumatoid arthritis[J]. Chinese Journal of Modern Applied Pharmacy, 2019, 36(4): 471-474.
10	HAN R F, GAO J, WANG L M, et al. microRNA-146a negatively regulates inflammation via the IRAK1/TRAF6/NF-κB signaling pathway in dry eye[J]. Sci Rep, 2023, 13(1): 11192.
11	ZHANG Y S, LIANG X J, BAO X F, et al. Toll-like receptor 4 (TLR4) inhibitors: current research and prospective[J]. Eur J Med Chem, 2022, 235: 114291.
12	PENG Y Q, CHEN B Q, SHENG X W, et al. The genetic association between TLR-1, -2, -4, and -6 gene polymorphisms and rheumatoid arthritis susceptibility in a Chinese Han population[J]. Genet Test Mol Biomarkers, 2022, 26(3): 140-145.
13	KONG L L, WANG L F, ZHAO Q, et al. Rhodojaponin Ⅱ inhibits TNF-α-induced inflammatory cytokine secretion in MH7A human rheumatoid arthritis fibroblast-like synoviocytes[J]. J Biochem Mol Toxicol, 2020, 34(10): e22551.
14	GUO F, WU S. Antisense IRAK-1 oligonucleotide blocks activation of NF-kappa B and AP-1 induced by IL-18[J]. Immunopharmacology, 2000, 49(3): 241-246.
15	GILYAZOVA I, ASADULLINA D, KAGIROVA E, et al. miRNA-146a: a key player in immunity and diseases[J]. Int J Mol Sci, 2023, 24(16): 12767.
16	TAVASOLIAN F, HOSSEINI A Z, SOUDI S, et al. miRNA-146a improves immunomodulatory effects of MSC-derived exosomes in rheumatoid arthritis[J]. Curr Gene Ther, 2020, 20(4): 297-312.
17	KONG R N, GAO J, JI L M, et al. Iguratimod ameliorates rheumatoid arthritis progression through regulating miR-146a mediated IRAK1 expression and TRAF6/JNK1 pathway: an in vivo and in vitro study[J]. Clin Exp Rheumatol, 2021, 39(2): 289-303.
18	MORTAZAVI-JAHROMI S S, AHMADZADEH A, REZAIEYAZDI Z, et al. The role of β-d-mannuronic acid, as a new non-steroidal anti-inflammatory drug on expression of miR-146a, IRAK1, TRAF6, NF-κB and pro-inflammatory cytokines following a clinical trial in rheumatoid arthritis patients[J]. Immunopharmacol Immunotoxicol, 2020, 42(3): 228-236.
19	张晓苹, 李茹, 王世瑶, 等. 微小RNA-146a对类风湿关节炎患者滑膜成纤维细胞抑制作用的研究[J]. 中华风湿病学杂志, 2010, 14(4): 220-222.
	ZHANG X P, LI R, WANG S Y, et al. The inhibitory effect of miR-146a on rheumatoid arthritis synovial fibroblast[J]. Chinese Journal of Rheumatology, 2010, 14(4): 220-222.
20	胡秋侠, 李博, 吴系美, 等. 注射用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白治疗类风湿关节炎疗效及与尿沉渣中miR146a表达的相关性[J]. 中国药师, 2018, 21(8): 1413-1415.
	HU Q X, LI B, WU X M, et al. Efficacy of recombinant human tumor necrosis factor-α receptor Ⅱ: Fc in the treatment of rheumatoid arthritis and its correlation with miR-146a expression in urinary sediment[J]. China Pharmacist, 2018, 21(8): 1413-1415.

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

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

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

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

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