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J Zhejiang Univ (Med Sci)  2018, Vol. 47 Issue (4): 389-394    DOI: 10.3785/j.issn.1008-9292.2018.08.10
    
Vitamin D3 regulates mononuclear phagocyte polarization induced by serum from patients with ankylosing spondylitis
WANG Shengnan1(),LYU Wang1,LIN Suxian1,LU Yang1,JIANG Jianchang1,ZHU Xiaochun2,*()
1. Department of Rheumatism and Immunology, Wenzhou Third Clinical Institute of Wenzhou Medical University, Wenzhou People's Hospital, Wenzhou 325000, Zhejiang Province, China
2. Department of Rheumatism and Immunology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang Province, China
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Abstract  

Objective: To investigate the effect of vitamin D3 on polarization of monocyte macrophages induced by serum from patients with ankylosing spondylitis (AS). Methods: Twenty AS naïve patients and 20 healthy controls from Wenzhou People's Hospital during January 2016 and December 2017 were enrolled. The macrophages were differentiated from THP1 cells induced by phorbol 12-myristate 13-acetate (PMA), and then co-cultured with the serum from healthy subjects (control group) or AS patients. Vitamin D3 was added in the medium mixed with serum from AS patients. Flow cytometry was used to analyze the ratio of CD68 and CD206 positive cells, and RT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). Results: THP1 cells could be polarized into mononuclear-macrophages with the induction of PMA. The proportion of CD206 positive cells in AS-serum group was lower than that in the control group (t=9.434, P < 0.05), while the proportion of CD68 positive cells was higher than that in the control group (t=43.920, P < 0.05). The proportion of CD206 positive cells in vitamin D3 group was higher than that in AS-serum group (t=8.895, P < 0.05), while the proportion of CD68 positive cells was lower than that in AS-serum group (t=9.089, P < 0.05). mRNA expression of Arg-1 in AS-serum group was lower than that in the control group (t=8.899, P < 0.05), while mRNA expression of iNOS was higher than that in the control group (t=3.656, P < 0.05). mRNA expression of Arg-1 in vitamin D3 group was higher than that in AS-serum group (t=6.219, P < 0.05), while mRNA expression of iNOS was lower than that in AS-serum group (t=5.876, P < 0.05). Conclusion: Vitamin D3 can regulate the polarization of mononuclear macrophages for immunoregulation in patients with AS.



Key wordsSpondylitis, ankylosing/etiology      Cholecalciferol/pharmacology      Macrophages/drug effects      T-Lymphocytes, helper-inducer/drug effects     
Received: 23 March 2018      Published: 04 December 2018
CLC:  R392  
Corresponding Authors: ZHU Xiaochun     E-mail: 35571795@qq.com;ygmm16@163.com
Cite this article:

WANG Shengnan,LYU Wang,LIN Suxian,LU Yang,JIANG Jianchang,ZHU Xiaochun. Vitamin D3 regulates mononuclear phagocyte polarization induced by serum from patients with ankylosing spondylitis. J Zhejiang Univ (Med Sci), 2018, 47(4): 389-394.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2018.08.10     OR     http://www.zjujournals.com/med/Y2018/V47/I4/389


维生素D3对强直性脊柱炎患者血清诱导单核巨噬细胞极化的影响

目的: 研究维生素D3干预对强直性脊柱炎(AS)患者血清单核巨噬细胞极化的影响。方法: 选择2016年1月至2017年12月在温州市人民医院尚未治疗的AS初诊患者20例和健康志愿者20名。通过佛波酯诱导人单核-白血病细胞THP1分化为单核巨噬细胞,并与AS患者(AS对照组)或健康志愿者(健康对照组)的外周血血清联合培养;在含有AS外周血血清的混合培养基中加入维生素D3进行干预(维生素D3组)。流式细胞仪测定CD68和CD206+单核巨噬细胞的比例,RT-PCR测定精氨酸酶1(Arg-1)和诱导型一氧化氮合酶(iNOS)mRNA的表达水平。结果: 佛波酯可将THP1细胞诱导分化为单核巨噬细胞并表现出明显的细胞形态变化。AS对照组中CD206+细胞的比例低于健康对照组(t=9.434,P < 0.05),而CD68+细胞的比例高于健康对照组(t=43.920,P < 0.05);维生素D3组中CD206+细胞的比例高于AS对照组(t=8.895,P < 0.05),而CD68+细胞的比例低于AS对照组(t=9.089,P < 0.05)。AS对照组中Arg1 mRNA表达量低于健康对照组(t=8.899,P < 0.05),而iNOS mRNA表达量高于健康对照组(t=3.656,P < 0.05);维生素D3组Arg-1 mRNA表达量高于AS对照组(t=6.219,P < 0.05),而iNOS mRNA表达量低于AS对照组(t=5.876,P < 0.05)。结论: 维生素D3可能通过影响单核巨噬细胞的极化参与AS患者的免疫调节。


关键词: 脊柱炎, 强直性/病因学,  胆骨化醇/药理学,  巨噬细胞/药物作用,  T淋巴细胞, 辅助诱导/药物作用 
Fig 1 Phorbol ester induces THP1 cells differentiate into mononuclear macrophages
Fig 2 Expression of CD206 and CD68 in mononuclear macrophages in three groups
($\bar x \pm s$, %)
组别 n CD206+细胞 CD68+细胞
与健康对照组比较,*P<0.05;与AS对照组比较,#P<0.05.AS:强直性脊柱炎.
健康对照组 5 27.8±3.9 47.8±1.2
AS对照组 5 9.7±1.4* 63.1±1.7*
维生素D3组 5 26.1±3.2# 49.0±1.5#
Tab 1 Proportions of CD206+ and CD68+ cells in three groups
Fig 3 mRNA expression of Arg-1 and iNOS in three groups
[1]   KIM H W , LEE S H . Pathogenesis of ankylosing spondylitis[J]. Nat Rev Rheumatol, 2015, 22 (2): 399- 405
[2]   VANAKI N , ASLANI S , JAMSHIDI A et al. Role of innate immune system in the pathogenesis of ankylosing spondylitis[J]. Biomed Pharmacother, 2018, 105 130- 143
doi: 10.1016/j.biopha.2018.05.097
[3]   DI R M , MALAGUARNERA G , DE GREGORIO C et al. Immuno-modulatory effects of vitamin D3 in human monocyte and macrophages[J]. Cell Immunol, 2012, 280 (1): 36- 43
[4]   LANGE U , TEICHMANN J , STRUNK J et al. Association of 1.25 vitamin D3 deficiency, disease activity and low bone mass in ankylosing spondylitis[J]. Osteoporos Int, 2005, 16 (12): 1999- 2004
doi: 10.1007/s00198-005-1990-5
[5]   GOIE T H S , STEVEN M M , VAN DER LINDEN S M et al. Evaluation of diagnostic criteria for ankylosing spondylitis:a comparison of the Rome, New York and modified New York criteria in patients with a positive clinical history screening test for ankylosing spondylitis[J]. Br J Rheumatol, 1985, 24 (3): 242- 249
doi: 10.1093/rheumatology/24.3.242
[6]   DAIGNEAULT M, PRESTON J A, MARRIOTT H M, et al. The identification of markers of macrophage differentiation in PMA-stimulated THP-1 cells and monocyte-derived macrophages[J/OL]. PLoS One, 2010, 5(1): e8668.
[7]   BRAUN J , BOLLOW M , NEURE L et al. Use of immunohistologic and in situ hybridization techniques in the examination of sacroiliac joint biopsy specimens from patients with ankylosing spondylitis[J]. Arthritis Rheum, 1995, 38 (4): 499- 505
doi: 10.1002/(ISSN)1529-0131
[8]   ZHAO S , DUFFIELD S J , MOOTS R J et al. Systematic review of association between vitamin D levels and susceptibility and disease activity of ankylosing spondylitis[J]. Rheumatology (Oxford), 2014, 53 (9): 1595- 1603
doi: 10.1093/rheumatology/keu042
[9]   VANDOOREN B , NOORDENBOS T , AMBARUS C et al. Absence of a classically activated macrophage cytokine signature in peripheral spondylarthritis, including psoriatic arthritis[J]. Arthritis Rheum, 2009, 60 (4): 966- 975
doi: 10.1002/art.v60:4
[10]   BAETEN D , DEMETTER P , CUVELIERC A et al. Macrophages expressing the scavenger receptor CD163:a link between immune alterations of the gut and synovial inflammation in spondyloarthropathy[J]. J Pathol, 2002, 196 (3): 343- 350
[11]   BISWAS S K , ALLAVENA P , MANTOVANI A . Tumor-associated macrophages:functional diversity, clinical significance, and open questions[J]. Semin Immunopathol, 2013, 35 (5): 585- 600
doi: 10.1007/s00281-013-0367-7
[12]   MARTINEZ F O , HELMING L , GORDON S . Alternative activation of macrophages:an immunologic functional perspective[J]. Annu Rev Immunol, 2009, 27 451- 483
doi: 10.1146/annurev.immunol.021908.132532
[13]   CHEN Y Z , ZHANG J , GE X et al. Vitamin D receptor inhibits nuclear factor κB activation by interacting with IκB kinase β protein[J]. J Biol Chem, 2013, 288 (27): 19450- 19458
doi: 10.1074/jbc.M113.467670
[14]   OLSON K C , KULLING P M , OLSON T L et al. Vitamin D decreases STAT phosphorylation and inflammatory cytokine output in T-LGL leukemia[J]. Cancer Biol Ther, 2017, 18 (5): 290- 303
doi: 10.1080/15384047.2016.1235669
[15]   ZHANG Y , LEUNG D Y , RICHERS B N et al. Vitamin D inhibits monocyte/macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1[J]. J Immunol, 2012, 188 (5): 2127- 2135
doi: 10.4049/jimmunol.1102412
[16]   MANTOVANI A , SICA A , LOCATI M . Macrophage polarization comes of age[J]. Immunity, 2005, 23 (4): 344- 346
doi: 10.1016/j.immuni.2005.10.001
[17]   ZHANG X L , ZHOU M , GUO Y F et al. 1, 25-dihydroxyvitamin D3 promotes high glucose-induced M1 macrophage switching to M2 via the VDR-PPARγ signaling pathway[J]. Biomed Res Int, 2015, 2015 (9): 1- 14
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