外周血单个核细胞中整合素α4对类风湿关节炎中的诊断价值及其生物学功能

doi:10.3969/j.issn.1673-8640.2026.10.011

摘要/Abstract

摘要：

目的探讨整合素α4（ITGA4）诊断类风湿关节炎（RA）的价值，并采用RA患者滑膜成纤维细胞（RA-FLS）探讨其生物学功能。方法选取2023年3月—2024年12月无锡市第九人民医院RA患者85例（RA组）和健康体检者100名（正常对照组）。收集所有研究对象的临床资料，并检测外周血单个核细胞（PBMC）中ITGA4的表达情况。分离并培养来源于RA患者的RA-FLS细胞，以甲氨蝶呤作为对照，使用ITGA4抑制剂（Carotegrast）处理细胞，检测Carotegrast干预后RA-FLS细胞的增殖、凋亡、周期、转移侵袭情况和白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）的表达情况。采用受试者工作特征（ROC）曲线评价关键基因对RA的诊断效能。采用Spearman分析评估ITGA4相对表达量与疾病活动指数28（DAS28）-类风湿因子（ESR）评分的相关性。结果 RA组PBMC中ITGA4相对表达量显著高于正常对照组（P<0.01）。ITGA4相对表达量诊断RA的曲线下面积（AUC）为0.672。ITGA4相对表达量与DAS28-ESR评分呈正相关（r=0.266 4，P=0.002 3）。Carotegrast干预后，RA-FLS的细胞增殖、细胞周期和迁移侵袭能力均受到抑制，细胞凋亡被促进。Carotegrast可有效抑制RA-FLS细胞中IL-1β、IL-6和TNF-α的表达。结论 RA患者PBMC中ITGA4表达显著升高，且与RA严重程度相关。ITGA4或可作为潜在的RA诊疗靶点。

关键词: 整合素α4, 外周血单个核细胞, 类风湿关节炎

Abstract:

Objective To determine the expression of integrin alpha 4 in rheumatoid arthritis（RA） patients and investigate its biological functions using rheumatoid arthritis fibroblast-like synoviocyte（RA-FLS）. Methods From March 2023 to December 2024，85 RA patients（RA group） and 100 healthy subjects（healthy control group） from Wuxi Ninth People's Hospital were enrolled. The clinical data were collected，and the expression of ITGA4 in peripheral blood mononuclear cells（PBMC） of RA patients was determined. RA-FLS cells of RA patients were isolated and cultured. Methotrexate was used as a control，and ITGA4 inhibitor（Carotegrast） was used to treat the cells. Changes in cell proliferation，apoptosis，cell cycle，metastasis and invasion，and the expression levels of interleukin（IL）-1β，IL-6 and tumor necrosis factor-alpha（TNF-α） in RA-FLS cells after Carotegrast intervention were determined. Receiver operating characteristic（ROC） curve was used to evaluate the diagnostic efficacy of key genes for RA. Spearman analysis was used to evaluate the correlation between the relative expression level of ITGA4 and disease activity score in 28 joints（DAS28）-erythrocyte sedimentation rate（ESR） score. Results The relative expression level of ITGA4 in PBMC of RA group was higher than that of healthy control group（P<0.01）. The area under curve（AUC） of ITGA4 relative expression level for diagnosing RA was 0.672. The relative expression level of ITGA4 was positively correlated with DAS28-ESR score（r=0.266 4， P=0.002 3）. After Carotegrast intervention，the cell proliferation，cell cycle and migration invasion ability of RA-FLS were inhibited，and cell apoptosis was promoted. Carotegrast could effectively inhibit the expression of IL-1β，IL-6 and TNF-α in RA-FLS cells. Conclusions The expression of ITGA4 in PBMC of RA patients is increased and is related to the severity of RA. ITGA4 may become a potential target for the diagnosis and treatment of RA.

Key words: Integrin alpha 4, Peripheral blood mononuclear cell, Rheumatoid arthritis

中图分类号:

R446.1

程国伟, 刘庆阳, 王秋波. 外周血单个核细胞中整合素α4对类风湿关节炎中的诊断价值及其生物学功能[J]. 检验医学, 2026, 41(10): 1004-1011.

CHENG Guowei, LIU Qingyang, WANG Qiubo. Diagnostic value and biological function of integrin alpha 4 in peripheral blood mononuclear cells in rheumatoid arthritis[J]. Laboratory Medicine, 2026, 41(10): 1004-1011.

图/表 11

参考文献 29

[1]	Jang S, Kwon E J, Lee J J. Rheumatoid arthritis:pathogenic roles of diverse immune cells[J]. Int J Mol Sci, 2022, 23(2).
[2]	Chastney M R, Conway J R W, Ivaska J. Integrin adhesion complexes[J]. Curr Biol, 2021, 31(10):R536-R542.
[3]	Aman J, Margadant C. Integrin-dependent cell-matrix adhesion in endothelial health and disease[J]. Circ Res, 2023, 132(3):355-378. DOI PMID
[4]	Rossi B, Dusi S, Angelini G, et al. Alpha4 beta7 integrin controls Th17 cell trafficking in the spinal cord leptomeninges during experimental autoimmune encephalomyelitis[J]. Front Immunol, 2023, 14:1071553. DOI URL
[5]	Calma A D, Young S, Sandbach J, et al. Targeting alpha-4 integrin with natalizumab for intermediate uveitis associated with multiple sclerosis[J]. Mult Scler J Exp Transl Clin, 2024, 10(4):20552173241301034.
[6]	Matsouka K, Watanabe M, Ohmori T, et al. AJM300(carotegrast methyl),an oral antagonist of alpha4-integrin,as induction therapy for patients with moderately active ulcerative colitis:a multicentre,randomised,double-blind,placebo-controlled,phase 3 study[J]. Lancet Gastroenterol Hepatol, 2022, 7(7):648-657. DOI URL
[7]	Alsadhan A, Chen J, Gaglione E M, et al. CD49d expression identifies a biologically distinct subtype of chronic lymphocytic leukemia with inferior progression-free survival on BTK inhibitor therapy[J]. Clin Cancer Res, 2023, 29(18):3612-3621. DOI URL
[8]	中华医学会风湿病学分会. 2018中国类风湿关节炎诊疗指南[J]. 中华内科杂志, 2018, 57(4):42-51.
[9]	Prevoo M L, Van'T Hof M A, Kuper H H, et al. Modified disease activity scores that include twenty-eight-joint counts. Development and validation in a prospective longitudinal study of patients with rheumatoid arthritis[J]. Arthritis Rheum, 1995, 38(1):44-48. DOI URL
[10]	Van Riel P L, Renskers L. The disease activity score(DAS) and the disease activity score using 28 joint counts(DAS28) in the management of rheumatoid arthritis[J]. Clin Exp Rheumatol, 2016, 34(5Suppl 101): S40-S44.
[11]	Wu Z, Ma D, Yang H, et al. Fibroblast-like synoviocytes in rheumatoid arthritis:surface markers and phenotypes[J]. Int Immunopharmacol, 2021, 93:107392. DOI URL
[12]	Alivernini S, Firesteun G S, Mcinnes I B. The pathogenesis of rheumatoid arthritis[J]. Immunity, 2022, 55(12):2255-2270. DOI PMID
[13]	Venetsanopoulou A I, Alamanos Y, Voulgari P V, et al. Epidemiology of rheumatoid arthritis:genetic and environmental influences[J]. Expert Rev Clin Immunol, 2022, 18(9):923-931. DOI URL
[14]	Cush J J. Rheumatoid arthritis:early diagnosis and treatment[J]. Rheum Dis Clin North Am, 2022, 48(2):537-547. DOI URL
[15]	Sepriano A, Kerschbaumer A, Bergstra S A, et al. Safety of synthetic and biological DMARDs:a systematic literature review informing the 2022 update of the EULAR recommendations for the management of rheumatoid arthritis[J]. Ann Rheum Dis, 2023, 82(1):107-118. DOI URL
[16]	Polman C H, O'Connor P W, Havrdova E, et al. A randomized,placebo-controlled trial of natalizumab for relapsing multiple sclerosis[J]. N Engl J Med, 2006, 354(9):899-910. DOI URL
[17]	Feagan B G, Greenberg G R, Wild G, et al. Treatment of ulcerative colitis with a humanized antibody to the alpha4beta7 integrin[J]. N Engl J Med, 2005, 352(24):2499-2507. DOI URL
[18]	Khoy K, Mariotte D, Defer G, et al. Natalizumab in multiple sclerosis treatment:from biological effects to immune monitoring[J]. Front Immunol, 2020, 11:549842. DOI URL
[19]	Ohmori T. Real-world effectiveness and safety of carotegrast methyl in Japanese patients with moderately active ulcerative colitis[J]. Inflamm Intest Dis, 2024, 9(1):271-282. DOI PMID
[20]	Chu H W, Schaunaman N. Very late antigen-4:a novel receptor for club cell secretory protein 16 to control inflammation[J]. Am J Respir Crit Care Med, 2021, 203(11):1340-1341. DOI URL
[21]	Savino W, Chaves B, Bonomo A C, et al. Integrin-directed antibody-based immunotherapy:focus on VLA-4[J]. Immunother Adv, 2021, 1(1):ltab002.
[22]	Barbadillo C, Ag A, Salas C, et al. Anti-integrin immunotherapy in rheumatoid arthritis:protective effect of anti-alpha 4 antibody in adjuvant arthritis[J]. Springer Semin Immunopathol, 1995, 16(4):427-436. PMID
[23]	Birner U, Issekutz T B, Walter U, et al. The role of alpha(4) and LFA-1 integrins in selectin-independent monocyte and neutrophil migration to joints of rats with adjuvant arthritis[J]. Int Immunol, 2000, 12(2):141-150. PMID
[24]	Dhillon S. Carotegrast methyl:first approval[J]. Drugs, 2022, 82(9):1011-1016. DOI
[25]	Takeuchi E, Tanaka T, Umemoto E, et al. VLA-4-dependent and -independent pathways in cell contact-induced proinflammatory cytokine production by synovial nurse-like cells from rheumatoid arthritis patients[J]. Arthritis Res, 2002, 4(6):R10.
[26]	Kondo N, Kuroda T, Kobayashi D. Cytokine networks in the pathogenesis of rheumatoid arthritis[J]. Int J Mol Sci, 2021, 22(20).
[27]	Ran Y, Dong Y, LI Y, et al. Mesenchymal stem cell aggregation mediated by integrin alpha4/VCAM-1 after intrathecal transplantation in MCAO rats[J]. Stem Cell Res Ther, 2022, 13(1):507. DOI PMID
[28]	Dong M, Wang W, Wu W, et al. A novel surface marker CD49d promotes TNF expression in oyster agranulocytes by mediating the MAPK pathway[J]. Fish Shellfish Immunol, 2024, 151:109702. DOI URL
[29]	Takeda T, Tsubak M, Genno S, et al. CD49d and CD49e induce cell adhesion-mediated drug resistance through the nuclear factor-kappaB pathway in Burkitt lymphoma[J]. J Physiol Pharmacol, 2020, 71(4).

基因名称	正向引物（5´~3´）	反向引物（5´~3´）
ITGA4	GAACCGATGGCTCCTAGTGG	GAAAGTGTGACCCCCAACCA
TNF-α	GTGCTTGTTCCTCAGCCTCT	ATGGGCTACAGGCTTGTCAC
IL-1β	CCAGCTACGAATCTCCGACC	TCGTGCACATAAGCCTCGTT
IL-6	AGAGGCACTGGCAGAAAACA	GCTCTGGCTTGTTCCTCACT
GAPDH	GTCAAGGCTGAGAACGGGAA	AAATGAGCCCCAGCCTTCTC

基因名称	正向引物（5´~3´）	反向引物（5´~3´）
ITGA4	GAACCGATGGCTCCTAGTGG	GAAAGTGTGACCCCCAACCA
TNF-α	GTGCTTGTTCCTCAGCCTCT	ATGGGCTACAGGCTTGTCAC
IL-1β	CCAGCTACGAATCTCCGACC	TCGTGCACATAAGCCTCGTT
IL-6	AGAGGCACTGGCAGAAAACA	GCTCTGGCTTGTTCCTCACT
GAPDH	GTCAAGGCTGAGAACGGGAA	AAATGAGCCCCAGCCTTCTC

项目	例数	年龄/岁	性别		BMI/（kg·m^-2）	RF/ （U·mL^-1）	抗CCP抗体/（U·mL^-1）	ESR/（mm·h^-1）	病程/年	DAS28评分/分	使用DMARD/[例（%）]
项目	例数	年龄/岁	男/例	女/例	BMI/（kg·m^-2）	RF/ （U·mL^-1）	抗CCP抗体/（U·mL^-1）	ESR/（mm·h^-1）	病程/年	DAS28评分/分	使用DMARD/[例（%）]
RA组	85	58.04±1.33	33	52	23.09±0.43	28.54±6.33	8.42±2.42	20.61±5.87	4.20±1.03	3.17±0.95	76
正常对照组	100	55.53±2.61	32	68	22.33±0.53	7.8±1.03	1.62±0.76
统计值		1.55	0.288		2.43	28.74	24.63
P值		0.54	0.34		0.76	<0.001	<0.001

项目	例数	年龄/岁	性别		BMI/（kg·m^-2）	RF/ （U·mL^-1）	抗CCP抗体/（U·mL^-1）	ESR/（mm·h^-1）	病程/年	DAS28评分/分	使用DMARD/[例（%）]
项目	例数	年龄/岁	男/例	女/例	BMI/（kg·m^-2）	RF/ （U·mL^-1）	抗CCP抗体/（U·mL^-1）	ESR/（mm·h^-1）	病程/年	DAS28评分/分	使用DMARD/[例（%）]
RA组	85	58.04±1.33	33	52	23.09±0.43	28.54±6.33	8.42±2.42	20.61±5.87	4.20±1.03	3.17±0.95	76
正常对照组	100	55.53±2.61	32	68	22.33±0.53	7.8±1.03	1.62±0.76
统计值		1.55	0.288		2.43	28.74	24.63
P值		0.54	0.34		0.76	<0.001	<0.001

项目	AUC（95%CI）	最佳临界值	敏感性/%	特异性/%	Youden指数
ITGA4 mRNA	0.672（0.596~0.748）	1.017	61.2	70.0	0.312
RF	0.840（0.782~0.898）	9.703	74.7	82.7	0.574
抗CCP抗体	0.720（0.649~0.791）	4.946	52.9	94.0	0.469