VITAMIN D: A NOVEL

THERAPEUTIC APPROACH FOR

KELOID, ANIN VITROANALYSIS
G.Y. Zhang; T. Cheng; Q. Luan; T. Liao; C.L. Nie; X.
Zheng; X.G. Xie; W.Y. Gao

Posted: 05/12/2011; The British Journal of

Dermatology.2011;164(4):729-737.2011Blackwell
Publishing

BACKGROUND

Vitamin D and its metabolites play an important

role in calcium homeostasis, bone remodelling,
hormone secretion, cell proliferation and
differentiation. Recent studies also suggest a
beneficial role of vitamin D in slowing the
progression of tissue fibrosis. However, their effects
on dermal fibrosis and keloids are unknown

OBJECTIVES

To investigate the effect of 1,25-dihydroxyvitamin

D3 (1,25D) in the pathogenesis of tissue fibrosis by
keloid fibroblasts (KFs)

METHODS

KFs were cultured and exposed to different

concentrations of 1,25D in the presence or absence of
transforming growth factor (TGF)-1. KF phenotypes
and protein production were analysed by real-time
reverse transcriptase-polymerase chain reaction,
Western blot, immunofluorescence and multiplex
enzyme-linked immunosorbent assay techniques.
Collagen synthesis was evaluated by measuringHproline incorporation. The effect of 1,25D on cell
proliferation and viability was evaluated by Formazan
assay, proliferating cell nuclear antigen expression and
the colorimetric conversion of 3-[4, 5-dimethylthiazol2-yl]-2, 5-diphenyltetrazolium bromide.

RESULTS

We confirmed the presence of vitamin D receptors

(VDRs) in cultured keloid fibroblasts. Fibroblasts
transfected with a vitamin D response element
reporter construct and exposed to the active vitamin D
metabolite 1,25D showed increased promoter activity
indicating VDR functionality in these cells. Incubation
of KFs with 1,25D suppressed TGF-1-induced collagen
type I, fibronectin and -smooth muscle actin
expression. 1,25D also modulated plasminogen
activator inhibitor-1 and matrix metalloproteinase-9
expression induced by TGF-1. Interestingly, 1,25D
induced hepatocyte growth factor mRNA expression
and protein secretion in keloid fibroblasts.

CONCLUSIONS

This study highlights key mechanistic pathways

through which vitamin D decreases fibrosis, and
provides a rationale for studies to test vitamin
D supplementation as a preventive and/or early
treatment strategy for keloid and related
fibrotic disorders.

DISCUSSION

Although clinical research has shown that vitamin D

can inhibit the formation of skin fibrosis, e.g.
systemic sclerosis,the real role of vitamin D in skin
remains largely unexplored
We hypothesized that vitamin D influences
profibrogenic processes by targeting normal
fibroblasts and KFs. In the present study, we
demonstrated VDR expression in normal skin and
keloid-derived fibroblasts. Furthermore, we found
that VDR localizes to the nucleus, consistent with its
role as a nuclear receptor, and that it is functional
as KFs transfected with a VDRE fused to a luciferase
reporter showed increased expression of the
transfected gene when exposed to 1,25D

As expected, TGF-1 increased secretion of matrix

proteins collagen type I, FN and -SMA and these
effects were significantly inhibited by 1,25D. These
observations are important for the following
reasons. Firstly, they confirm that KFs are indeed
capable of recognizing 1,25D through functional
VDRs. Secondly, another important observation
relates to the fact that 1,25D was capable of
inhibiting the TGF-1-mediated tissue remodelling
responses in KFs. Although quite limited, much of
the work on the role of vitamin D in TGF-1-related
fibrosis has been studied, not unexpectedly, in cells
or organs known to be vitamin D targets.

Activation of vitamin D pathways has been observed

to decrease matrix expression and/or affect TGF-1
signalling in experimental renal fibrosis,ultraviolet
radiation-induced skin fibrosis,and viral myocarditis
Incubation with 1,25D increased the expression of
MMP-9, a potent collagen breakdown inducer, which
cleaves collagen at a single site and renders it
susceptible to degradation by other MMPs and
proteases leading to a reversion of early fibrosis,and
decreased the PAI-1 expression. This demonstrates
that modulating PAI-1 may contribute to the process
of keloid formation and tissue fibrosis via enhancing
the antiproteolytic activity of the ECM metabolism
equilibrium towards a state of impaired degration

HGF has recently been confirmed as an antifibrotic

cytokine that prevents the genesis and progression
of fibrotic lesions in keloids
1,25D also regulated HGF production by TGF-1 in
KFs, which was confirmed by previous research.This
establishes that HGF may act as a downstream
effect that mediates the action of 1,25D in the
keloid. These observations provide significant
insights into understanding the mechanism by which
1,25D ameliorates skin tissue fibrosis

One intriguing observation is that VDR interacts

directly with Smad3, thereby regulating TGF1/Smad signalling. However, such interaction
between VDR and Smad3 results in an enhancement
of TGF-1 signalling, and therefore cannot account
for an inhibitory effect of 1,25D on TGF-1 action
Thus, the observation in the current study of the
antifibrotic effect in keloid by the active form of
vitamin D suggests a potential therapeutic effect
for keloid

In conclusion, on the basis of ourin vitroevidence

we can state that 1,25D-targeted inhibition of ECM
and induced HGF secretion may be an appropriate
therapeutic strategy for the management of keloid.
However, the precise molecular mechanisms of the
effects of 1,25D still need to be explored in KFs