Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells

Abstract

BackgroundLow levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells.MethodsHFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-1β (IL-1β) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D (1,25(OH)2D) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction.ResultsIL-1β significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and 1,25(OH)2D (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and 1,25(OH)2D.ConclusionVitamin D, 25(OH)D, and 1,25(OH)2D play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-1β stimulated MMP-9 production and conversion to its active form but also inhibiting IL-1β inhibition on TIMP-1 and TIMP-2 production.

Molecular actions of vitamin D in reproductive cell biology

“…However, although increased expression of MMP-2 and -9 was observed, the cellular signalling mechanism driving this phenotype was also not explored, again demonstrating the limited knowledge in this area. Particularly, considering that other studies in human lung fibroblasts and uterine fibroid cells showed that VitD metabolites actually inhibited MMP-2 and -9 expression by modulating cytokine expression (Halder et al 2013, Kim et al 2014, which are key regulators of immune cell infiltration. Nonetheless, these conflicting effects may be explained by the complex mechanism of VDR and VDRE regulation as outlined earlier.…”

SPLICELECT™: an adaptable cell surface display technology based on alternative splicing allowing the qualitative and quantitative prediction of secreted product at a single-cell level

“…46,[73][74][75][76] In addition to polyadenylation signals, so-called "terminators", such as the human gastrin terminator, have been described to play an important role in transcription terminator. 77 In our AS construct, neither addition of polyadenylation signals or gastrin terminator in the intron, nor addition of the gastrin terminator at the end of the expression cassette had an effect on secretion or cell surface display.…”

“…The gastrin terminator was described to detach the transcription machinery from the DNA. 77 Added at the end of the expression cassette after the polyadenylation signal, it may affect splicing, but the addition of the gastrin terminator at the end of the expression cassette had no significant impact on either surface staining or titer (data not shown).…”

Environmental Influencers, MicroRNA, and Multiple Sclerosis

“…48,49 Moreover, it can inhibit chemokine and cytokine expression, monocyte accumulation, and inducible nitric oxide synthaseaccumulation in the CNS. 50,51 It can also suppress the expression and activity of Matrix metalloproteinases-2 and-9 (MMP-2 and MMP-9), 52 which are associated with a risk of MS. 53 Although many investigators have demonstrated the effect of vitamin D supplementation on MS or experimental autoimmune encephalomyelitis (EAE) models, the exact modulation pathways remain elusive. Several theories have been proposed to explain the modulation effects of vitamin D on MS pathophysiology: (1) Vitamin D supplementation could increase the vitamin D receptor (VDR)-binding sites across the genome, 25 which were found to be enriched in genes previously associated Figure 1.…”