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30 January 2017 : Laboratory Research

1,25-Dihydroxyvitamin D3 Does Not Affect MicroRNA Expression When Suppressing Human Th17 Differentiation

Jian Huang12ABCDEFG, Zibin Liang12BF, Ying Kuang12BF, Fujie Jia12BF, Yaqi Yang12BF, Miaomiao Kang12BF, Muke Xie12BF, Feng Li12ABCDEFG*

DOI: 10.12659/MSM.898824

Med Sci Monit 2017; 23:535-541

Abstract

BACKGROUND: Vitamin D is an import regulator of T helper 17 (Th17) differentiation, but our understanding of the underlying mechanisms remains limited. In the present study, we aimed to detect the expression levels of microRNAs (miRNAs) during human Th17 differentiation and evaluate the effects of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), the bioactive form of vitamin D, on Th17 differentiation and miRNA expression.

MATERIAL AND METHODS: We cultured human peripheral blood mononuclear cells (PBMC) in vitro and activated them with anti-CD3 and anti-CD28 antibodies in the presence of Th17-promoting cytokines interleukin (IL)-23, IL-1β, TGF-β1, and IL-6 for 72 hours. 1,25(OH)2D3 was added to the medium at a final concentration of 100 nM on day 0. The production of IL-17A in culture medium was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of miRNAs during Th17 differentiation were determined by quantitative polymerase chain reaction (qPCR).

RESULTS: Six miRNAs were found to be dysregulated during human Th17 differentiation. Of these miRNAs, hsa-miR-155 was significantly up-regulated (median fold change: 3.61, P<0.05), whereas hsa-miR-20b, hsa-miR-21, hsa-miR-181a, hsa-miR-210, and hsa-miR-301a were significantly down-regulated (median fold change: 0.44, 0.37, 0.18, 0.15, and 0.26, respectively, P<0.05). 1,25(OH)2D3 treatment significantly decreased IL-17A production (median [interquartile range], 745.7 [473.5] pg/mL vs. 2535.4 [2153.3] pg/mL, P<0.05). However, expression of these miRNAs was not changed after 1,25(OH)2D3 treatment.

CONCLUSIONS: 1,25(OH)2D3 suppressed human Th17 differentiation without affecting miRNA expression.

Keywords: Cell Differentiation - drug effects, Calcitriol - pharmacology, Cells, Cultured, Cytokines - pharmacology, Enzyme-Linked Immunosorbent Assay, Interleukin-17 - biosynthesis, Leukocytes, Mononuclear - metabolism, MicroRNAs - metabolism, Th17 Cells - metabolism, Vitamins - pharmacology

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750