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Vitamin K was discovered early last century at the same time as the vitamin K-antagonists. For many years the role of vitamin K was solely ascribed to coagulation and coagulation was thought to be involved only at the venous blood side. This view has dramatically changed with the discovery of vitamin K-dependent proteins outside the coagulation cascade and the role of coagulation factors at the arterial side. Vitamin K-dependent proteins are involved in the regulation of vascular smooth muscle cell migration, apoptosis, and calcification. Vascular calcification has become an important independent predictor of cardiovascular disease. Vitamin K-antagonists induce inactivity of inhibitors of vascular calcification, leading to accelerated calcification. The involvement of vitamin K-dependent proteins such as MGP in vascular calcification make that calcification is amendable for intervention with high intake of vitamin K. This review focuses on the effect of vitamin K-dependent proteins in vascular disease.
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Hämostaseologie 4/2011
1 © Schattauer 2011Review
Role of vitamin K-dependent
proteins in the arterial vessel wall
M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers
Department of Biochemistry, CARIM, Maastricht University, the Netherlands
Vitamin K, vitamin K-antagonists, MGP,
vascular calcification
Vitamin K was discovered early last century at
the same time as the vitamin K-antagonists.
For many years the role of vitamin K was
solely ascribed to coagulation and coagu-
lation was thought to be involved only at the
venous blood side. This view has dramatically
changed with the discovery of vitamin K-de-
pendent proteins outside the coagulation cas-
cade and the role of coagulation factors at the
arterial side. Vitamin K-dependent proteins
are involved in the regulation of vascular
smooth muscle cell migration, apoptosis, and
calcification. Vascular calcification has be-
come an important independent predictor of
cardiovascular disease. Vitamin K-antagonists
induce inactivity of inhibitors of vascular cal-
cification, leading to accelerated calcification.
The involvement of vitamin K-dependent pro-
teins such as MGP in vascular calcification
make that calcification is amendable for inter-
vention with high intake of vitamin K. This re-
view focuses on the effect of vitamin K-de-
pendent proteins in vascular disease.
Correspondence to:
Leon J. Schurgers, PhD
Dept. of Biochemistry, Maastricht University
PO Box 616, 6200 MD Maastricht
The Netherlands
Tel. +31/43/388 16 80, Fax +31/43/388 41 59
Vitamin K, Vitamin-K-Antagonisten, MGP,
vaskuläre Kalzifikation
Vitamin K wurde im frühen vergangenen Jahr-
hundert zur gleichen Zeit wie die Vitamin-
K-Antagonisten entdeckt. Über Jahre wurde
die Rolle von Vitamin K ausschließlich der
Blutgerinnung zugeschrieben und man glaub-
te, dass sich die Koagulation nur auf der venö-
sen Blutseite abspiele. Mit der Entdeckung Vi-
tamin-K-abhängiger Proteine außerhalb der
Gerinnungskaskade und der Bedeutung von
Gerinnungsfaktoren auf der arteriellen Seite
hat sich diese Sicht grundlegend geändert. Vi-
tamin-K-abhängige Proteine sind an der Re-
gulation der Migration glatter Gefäßmuskel-
zellen sowie an der Apoptose und Kalzifikati-
on beteiligt. Vaskuläre Kalzifikation ist ein
wichtiger unabhängiger Prognosefaktor für
kardiovaskuläre Erkrankungen. Vitamin-K-
Antagonisten induzieren eine Inaktivität von
Inhibitoren der vaskulären Kalzifikation und
führen so zu beschleunigter Verkalkung. Die
Beteiligung Vitamin-K-abhängiger Proteine
wie MGP an der vaskulären Kalzifikation er-
öffnet eine Möglichkeit zur therapeutischen
Intervention durch die Einnahme hoher Dosen
Vitamin K. Das Thema dieses Reviews ist die
Wirkung Vitamin-K-abhängiger Proteine auf
vaskuläre Erkrankungen.
Die Rolle Vitamin-K-abhängiger Proteine in der
arteriellen Gefäßwand
Hämostaseologie 2011; 31: ■■
received: May 17, 2011
accepted: June 6, 2011
prepublished online: June 29, 2011
Vitamin K
Vitamin K is a fat-soluble vitamin and be-
longs to the family of vitamins A, D and E.
For most people, vitamin K is the least
known vitamin and even regarded as the
Cinderella (1). After the discovery of vit-
amin K in the early days of last century
clinicians and scientists believed that blood
coagulation was the only physiological pro-
cess in which vitamin K played a role. It was
the Danish researcher Dam who discovered
that chickens fed a fat-free diet suffered
from serious bleedings. It took some time
to isolate the micronutrient responsible for
this action, and it is now known as vitamin
K after the German word “Koagulation.
In the 1970s it was discovered that vit-
amin K is a cofactor in the carboxylation
reaction. Simultaneously Stenflo et al. (2)
and Nelsestuen et al. (3) reported the dis-
covery of the unusual amino acid γ-carbo-
xyglutamate acid (Gla) in prothrombin as
the product of vitamin K action. These
groups independently identified the un-
equivocal role of vitamin K as a cofactor for
the post-translational carboxylation of glu-
tamate (Glu) residues. This carboxylation-
step is accomplished by an enzyme called
gamma glutamyl-carboxylase (GGCX) (4),
and requires a pro-peptide containing pro-
This process is driven by the oxidation
of reduced vitamin K into vitamin K-epox-
ide. The vitamin K-epoxide must be re-
cycled to vitamin K before it can be reused.
This reaction is catalyzed by the enzyme
vitamin K epoxide reductase (VKOR) (5,
6). In this way the efficiency of vitamin K is
very high:
One molecule vitamin K can assure some
500 carboxylation reactions.
Recently, the group of Oldenburg added a
new role for the VKOR enzyme in that the
subunit VKORC1L1 is responsible for
driving vitamin K-mediated intracellular
antioxidation pathways critical to cell sur-
vival (7).
Dietary vitamin K
Vitamin K is an essential dietary micro-
nutrient since man cannot synthesize it. Al-
though in our gut flora some bacteria pro-
duce large amounts of vitamin K2 (8) their
contribution to the vitamin K-status is
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2M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers: Vitamin K-dependent proteins
questionable (9). The recommended daily
intake for vitamin K1 is 1 μg/kg body
weight and this is solely based on blood co-
agulation (10).
With the discovery of vitamin K-de-
pendent proteins in extra-hepatic tissues –
such as bone and vessel wall this view
needs to be revised. Nutritional vitamin K
consists of two forms:
vitamin K1 and
vitamin K2 .
Vitamin K1 (also called phylloquinone) is
found in leafy green vegetables where it is
tightly bound to the chloroplast mem-
brane. This results in a poor absorption of
vitamin K1 from vegetables (11, 12) and
thus its contribution to the vitamin
K-status is overestimated.
Vitamin K2 (group name for the mena-
quinones) is found in fermented foods
such as cheese, sauerkraut and the Japanese
natto (12) which is derived from the bacte-
ria that are used for the fermentation pro-
cess. The absorption of vitamin K2 is much
better as compared to vi tamin K1 (13). The
difference between vitamin K1 and vitamin
K2 is related to the aliphatic side chain.
After being absorbed in the intestine vit-
amin K is transported by lipoproteins, as it
has no specific carrier protein. The differ-
ent lipophilicity of K1 and K2 may result in
substantial differences in plasma transport,
half life and delivery to target tissues (14)
Tab. 1 ).
Vitamin K-antagonists (VKA)
Vitamin K-antagonists are 4-hydroxy -
coumarin derivatives. VKA were discover-
ed in the early 1920s as a malady of cattle
involving fatal bleeding showing up almost
simultaneously in the area of Wisconsin
(15). It turned out that if cattle ate spoiled
hay a loss in the clotting power of the blood
and as a resultant internal haemorrhage oc-
curred, which usually became fatal. It was
Campbell who isolated the crystalline di-
coumarol in 1939 (16).
Dicoumarol was launched as rat poison
in the early 1940s. After an unsuccessful
suicide attempt of an US soldier it became a
drug to lower the coagulation tendency of
blood. VKA have been given to patients for
more than six decades and besides an in-
creased bleeding tendency they are
relatively safe. By the year 2000 VKA were
the 11
most prescribed drug in the United
States (17).
Worldwide VKA form the mostly used drug
for the treatment and prevention of throm-
boembolic events, including atrial fibril-
lation, deep venous thrombosis and artifi-
cial heart valves.
In the USA, warfarin (named after the Wis-
consin Alumni Research Federation) is the
most used VKA whereas in Europe also
phenprocoumon and acenocoumarol are
used (
Tab. 1 ).
The action of VKA is to block the VKOR
enzyme thereby rapidly exhausting vitamin
K tissue stores (18). VKA are used to pre-
vent thrombosis in patients at increased
risk for thrombosis. When patients are
over-anticoagulated with VKA this can be
reversed by a high amount of supplemental
vitamin K.
The liver has a very active antidotal
pathway for VKA, called the DT-diapho-
rase (19, 20). This enzyme is some 100-fold
less active in bone and vessel wall. There-
fore, VKA cause a pronounced vitamin
K-deficiency in extra-hepatic tissues (21,
22). With the knowledge of today – 16 vit-
amin K-dependent proteins are now
known, half of them synthesized by tissues
other than the liver – the use of VKA may
also induce unwanted side effects (
Ta b.
Vitamin K-dependent
It is now known that vitamin K-dependent
proteins constitute a family of 16 known
proteins with diverse functions, not only
involved in the haemostatic pathway
Tab. 2). We will summarize the vitamin
K-dependent proteins, in particular those
involved in vascular disease.
Coagulation proteins
The clotting factors II, VII, IX and X are es-
sential for the coagulation cascade and are
γ-carboxylated in the liver to be function-
ally active. They are well balanced by the
anticoagulant factors protein C, protein S
and protein Z. These vitamin K-dependent
proteins are mainly synthesized and γ-glu-
tamylcarboxylated in the liver, with the ex-
ception of proteins S which is synthesized
some 45% by endothelial cells (23). Only
recently it has been realized that coagu-
lation factors also play an important role in
inflammation (24, 25). Minute amounts of
the coagulation proteins prothrombin and
FVII are synthesized de novo in the vessel
wall (24). The inhibition of thrombin by
melagatran (direct FIIa inhibitor) reduced
atherosclerotic plaque size and features of
plaque vulnerability (26).
Protein S acts in the coagulation cascade
as a cofactor of activated protein C (APC)
in the degradation of FVa and FVIIIa.
Tab. 1 Vitamin K and vitamin K-antagonists
name (trivial name) primary source half-life (t1/2) in hours
vitamin K phylloquinone leafy green vegetables 3
menaquinone meat, eggs 1.5
natto, cheese > 70
cheese, curd, sauerkraut > 70
vitamin K
acenocoumarol (Sintrom) 10
dicoumarol (Warfarin) 50
phenprocoumon (Marcumar) 100
brodifacum (super coumarin) > 1500
-8, -9
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3 M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers: Vitamin K-dependent proteins
Besides its coagulation inhibiting proper-
ties, protein S mediates a variety of regula-
tory phenomena including apoptosis and
phagocytosis (27).
Phagocytosis of apoptotic cells is thought
to limit the inflammatory response (28).
Protein S has been identified as a factor re-
sponsible for stimulation phagocytosis of
apoptotic cells by macrophages (27). Addi-
tionally, protein S regulates the expression
and function of scavenger receptor A
(SR-A) on macrophages resulting in dim-
inished uptake of acetylated low density li-
poprotein (AcLDL) (29, 30).
Recently, a new function of protein S
was discovered in vessel wall development.
Mice with protein S deficiency die in utero
(31). Mutants in which protein S was de-
leted specifically in hepatocytes – thought
to be the major source of circulating pro-
tein S – were viable as adults due to the en-
dothelial synthesized protein S. Protein S
deleted in endothelial cells revealed that en-
dothelial cells synthesize some 45% of the
blood-borne protein S (23). Interestingly,
the deficiency in the vessel wall resulted in
impaired angiogenesis, independent of
protein C (23). Mice with heterozygous
protein C deficiency not only exhibit severe
coagulation response to endotoxin but also
have significant differences in their inflam-
matory response (32). Additionally, APC
has been found to inhibit endotoxin-in-
duced production of TNF-α, IL-1β, IL-6,
and IL-8 by cultured monocytes/macro-
phages (33).
Extrahepatic vitamin K-dependent
Within the arterial vessel wall vitamin
K-dependent proteins are synthesized with
functions not related to blood coagulation.
Growth arrest specific gene 6 protein
(Gas-6) is a vitamin K-dependent protein
produced by vascular smooth muscle cells
(VSMCs) and involved in a pleiotropic of
physiologically processes (34). Gas-6 is as-
sociated with binding to its receptor Axl,
stimulating the anti-apoptotic protein
bcl-2 and inhibiting the pro-apoptotic pro-
tein caspase-3. Son et al. (35) showed that
Gas-6-Axl signaling protects VSMCs from
calcification by inhibiting apoptosis. It has
been shown that apoptotic bodies may
form a nidus for calcification (36).
The vitamin K-dependent matrix Gla-pro-
tein (MGP) is regarded as the strongest in-
hibitor of vascular calcification (VC) and
produced by many cells, including VSMCs.
promotes VSMC differentiation,
antagonizes BMP (BMP2 and BMP4)
signaling and
prevents osteochondrogenic lineage
reprogramming of VSMCs.
Both a high local and circulating inactive
MGP was associated with significantly
more VC and cardiovascular death (37, 38).
The role of MGP was elucidated in MGP
null mice (39). These mice were born
normally, but all died within eight weeks
after birth from ruptures of the large vessels
due to their massive calcification and loss of
elasticity. Rescue experiments in MGP null
mice demonstrated that MGP acts locally in
the vascular tissue as restoration of MGP
expression in arteries completely rescued
the arterial mineralization phenotype,
whereas hepatic MGP expression, resulting
in high systemic MGP levels, did not (40).
The crucial role of vitamin K in the in-
hibition of VC became clear from experi-
ments in which VKA was administered to
experimental animals (41). In this model –
in which VKA was given in the presence of
vitamin K to prevent bleedings – all extra-
hepatic vitamin K-dependent proteins, in-
cluding MGP, were synthesized in their in-
active uncarboxylated form, resulting in
vascular calcifications within 2–4 weeks.
Recently, we found that the VKA treat-
Tab. 2 Vitamin K-dependent proteins
VKD protein in tissue of γ-carboxylation (ref.) function (ref.)
factor II
liver. limited extra-hepatic (24) coagulation, regulatory functions in
inflammation (24, 25, 26)
factor VII coagulation
factor IX liver
factor X
protein C liver (80) coagulation (25), anti-in flammatory
(32, 33)
protein S liver and endothelial cells (23) coagulation (25), anti-in flammatory,
phago cytosis and apoptosis (28–30)
Gla-rich protein regulator of mineralisation (83)
Gla proteins
unknown (84)
Gla proteins
most soft tissues
bone marrow mesenchymal
stromal cells, cadiomyocytes
unknown (85)
bone (86), myocardial (87)
protein Z liver (81) degradation of factor Xa (79)
other VKD proteins
MGP bone, cartilage, vascular tissue
and macrophages (38, 39, 57)
negative regulator of vascular
calcification (37–39)
Gas-6 VSMCs and endothelial cells TAM activating ligand (34)
osteocalcin EH: primarily osteoblasts (82) extra cellular matrix protein in bone
(78, 82)
TGF-α inducible
most soft tissues ECM protein
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4 M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers: Vitamin K-dependent proteins
ment also caused increased apoptosis in the
vascular media, which further supports the
relation between apoptosis and calcifi-
cation (36).
The role of vitamin K-dependent proteins
has also been studied in patient popu-
lations and it confirmed that treatment with
VKA induces excessive calcification of the
vascular arteries and aortic heart valves
Additionally, the duration of VKA treat-
ment seems to correlate with an increase in
vascular calcification.
Vascular calcification
VC is associated with increased cardiovas-
cular mortality and morbidity, and is re-
cognised as a strong and independent risk
factor for cardiovascular death (45–47).
The amount of VC, as measured and
quantified by multidetector computed to-
mography is an important predictor of
all-cause mortality,
vascular complications and
myocardial infarctions (45, 48, 49).
Patients with higher coronary artery cal-
cification scores were approximately ten
times more likely to have a cardiac event in
the next 3–5 years (50). Not only the pres-
ence of coronary artery calcification is pre-
dictive for cardiovascular outcome, also its
annual increase. It was shown that patients
with a calcification-progression over 15%
per year had a 17.2 fold increased risk of
myocardial infarction compared to pa-
tients without significant progression (51).
The amount of VC is even a stronger pre-
dictor than the Framingham risk score
(FRS) (52), a well accepted 10-year risk pre-
dictor for coronary vascular disease. Clini-
cally, VC causes stiffening of the vascular
arteries via elastic fiber and VSMC calcifi-
cation. The calcification may result in
decreased arterial compliance,
development of left ventricular hyper-
trophy and
decreased coronary perfusion leading to
an increased risk of fatal complications.
In spite of this, calcification of arteries has
been neglected and considered to be clini-
cally irrelevant. VC was regarded as an end-
stage passive process not amenable to
therapeutic intervention (53). However, re-
cent reports demonstrate that punctated
and spotty calcification in the athero-
sclerotic plaque influence stability
negatively and render the plaque vulner-
able to rupture (54, 55) (
Fig. 1).
VC is now appreciated as a complex and
actively regulated process involving cells
and proteins acting as catalysts and
inhibitors (56, 57).
Recruitment of macrophages in the athero-
sclerotic plaque and consequently their se-
cretion of inflammatory cytokines may
serve as a signal for intimal calcification.
Indeed Nadra et al. showed that basic cal-
cium phosphate crystals are taken up by
macrophages in vitro (58). This was associ-
ated with the secretion of the pro-inflam-
matory cytokines TNF-α, IL-β and IL-8.
Furthermore, Pazár et al. showed that basic
calcium phosphate induced macrophage
IL-1β secretion through activation of the
NLRP3 inflammasome (59, 60). Also
VSMCs can execute phagocytosis of cal-
cium crystals. Ewence et al. showed that
when VSMCs phagocytose calcium crystals
it might destabilize atherosclerotic plaques
by initiating inflammation and by causing
VSMC death (61) (
Fig. 2).
Detection of vascular calcification
VC ca n b e v isua li zed b y v ario us tech ni ques .
In the clinical setting, multidetector com-
puted tomography is often used and gener-
ates a quantitative calcium score, which is
used as a measure of atherosclerotic burden
(62, 63). VC is therefore a potent predictor
for cardiovascular events (64). Although
some research has linked the amount of
vascular calcium to a more stable plaque
phenotype (65) most studies identified
intimal calcification as predictor of a vul-
nerable plaque phenotype, in particular the
punctated “spotty calcification” (63, 66).
Indeed, finite element analysis implied that
macrocalcification in the plaque did not in-
crease plaque stress or rupture (55, 67)
Fig. 1 Detection of vascular calcification
a) in vivo micro-CT: calcified atherosclerotic plaque of an ApoE-/- mouse on western type diet for 6 months
b) histochemical staining (von Kossa) of intimal microcalcification of an ApoE-/- mouse on western type diet containing warfarin and vitamin K for 3 months
a) b)
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5 M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers: Vitamin K-dependent proteins
whereas small calcified spots in the athero-
sclerotic cap increased stress, sufficient for
causing plaque rupture.
This resulted in studies screening for regu-
lating mechanisms of VC by multimodality
imaging. Derlin et al. (68), used a combination
of positron emitted tomography (PET) and
CT to asses the cardiovascular risk in patients.
The use of
F sodium fluoride for imaging
calcified atherosclerotic plaques showed a
more frequent uptake of Na
F in patients
with a high-risk profile. However, there was a
weaker correlation with risk factors compared
to the calcified plaque burden (68).
Imaging of VC in mice was done using
bisphosphonate near-infrared conjugated
probes. Bisphosphonates strongly bind to
calcified structures in coronary arteries
(69, 70). However, bisphosphonate also
strongly accumulates in bone, which results
in a high background signal. Therefore,
these studies are only applied ex vivo. New
specific probes imaging microcalcification
can provide a platform to study the earliest
events associated with VC at the molecular
and cellular level.
The use of circulating biomarkers such
as MGP for detecting or screening VC is an
attractive possibility. Vitamin K-dependent
proteins have been associated with the ear-
liest calcification areas in the plaque (71). It
was the uncarboxylated form of MGP that
strongly correlated with both medial and
intimal calcification (71, 72). By measuring
circulating MGP isoforms it was shown
that the majority of the healthy population
have sub-optimal levels of vascular vitamin
K (73, 74). Preliminary data suggest that
some MGP conformations are associated
with aspects of cardiovascular disease (37,
38, 75, 76). Patients with high VC scores
display high levels of inactive MGP, es-
pecially dialysis patients.
This creates possibilities for targeting VC
with vitamin K. Indeed high intake of vit-
amin K has been shown to regress preformed
medial calcifications in a rat model (77).
Recently, we conducted a first pilot
study in dialysis patients showing that vit-
amin K supplementation markedly re-
duced the level in plasma (78) of
uncarboxylated prothrombin (pivka-II),
uncarboxylated osteocalcin (ucOC),
inactive MGP (dp-ucMGP).
Fig. 2 Mechanism of vascular calcification
1) Adaptation of VSMCs from a contractile to a synthetic phenotype as a
result of multiple stress factors. VSMCs start loading calcium resulting in
calcification of the media.
2, 3) Additionally, modified lipoprotein binding to macrophage scavenger
receptors (2) are phagocytosed and accumulation results in foam cell
formation (3).
4) Foam cells secrete pro-inflammatory cytokines that amplify the local in-
flammatory response (4) resulting in an accelerated calcium loading and
vesicle release by VSMCs. These vesicles form a nidus for calcification. The in-
creased inflammatory profile results in osteogenic differentiation of VSMCs.
Additionally, VSMCs lose their calcifying inhibitors (such as MGP) resulting in
an acceleration of calcification.
5) As a result, VSMCs undergo apoptosis releasing more apoptotic bodies,
accelerating the calcification process.
6, 7) Macrophages phagocytose calcium crystals (7) which induces
activation of the NLRP-3 inflammasome.
8) Subsequently cytokines (such as TNF-α, IL-β and IL-8) are released. Also
VSMCs phagocytose calcium crystals, which leads to enhanced apoptosis of
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6 M. L. L. Chatrou; C. P. Reutelingsperger; L. J. Schurgers: Vitamin K-dependent proteins
Effort must be directed towards retarding
or reversing the development of calcifi-
cation in the vasculature, especially in those
patients prone for vascular calcification
(i. e. chronic kidney disease, diabetes, car-
diovascular disease). In these patients the
treatment with vitamin K antagonists
should be reconsidered. Data suggest that
high vitamin K can induce regression of
VKA-induced vascular calcification (77).
Therefore, it is of importance to identify
patients with vascular disease and to evalu-
ate different strategies that are more effec-
tive in the prevention of
hypercoagulability as well as
vascular calcification.
New oral anticoagulants such as FIIa- and
FXa-inhibitors that specifically target one
protein in the coagulation cascade without
affecting vascular vitamin K-dependent
proteins may become the preferred choice.
Conflict of interest
The authors report no conflict of interests.
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... Néanmoins, une autre enzyme à activité Vitamine K Réductase VKR non inhibée par les AVK intervient à cette étape mais elle est non identifiée encore. La KH2 pourra redémarrer de nouveaux le cycle, ainsi, une seule molécule de vitamine K est capable de faire 500 carboxylations différentes (Chatrou et al., 2011) . Les enzymes citées ci-dessus sont détaillées dans la partie suivante. ...
Les pullulations de campagnols terrestres sont à l’origine de nombreux problèmes. Leur gestion implique différentes mesures mais surtout une lutte chimique par les antivitamines K (AVK). Ces molécules ciblent l’enzyme VKOR codée par le gène VKORC1. L’inhibition de cette enzyme provoque des hémorragies mortelles. L’utilisation intensive des AVK a mené à l’émergence d’une résistance de cible ainsi que des ecotoxicités documentées par plusieurs études. Au cours de ce travail, nous avons étudié la pharmacocinétique de la chlorophacinone et de la bromadiolone chez le campagnol. Nous tenons à préciser que le mélange actuel de bromadiolone utilisé dans la lutte est composé de 85% de l’isomères trans et de 15% de l’isomères cis. Notre étude nous a permis d’identifier une longue demi-vie de l’isomère trans de la bromadiolone contre une demi-vie beaucoup plus réduite pour la cis-bromadiolone et la chlorophacinone. Par conséquence, l’isomère trans est responsable de la persistance de la molécule chez le campagnol et ensuite de l’intoxication des espèces non cible. Nous nous sommes ensuite intéressés au phénomène de la résistance de cible aux AVK.Nous avons tout d’abord, participé à une étude de la résistance sur différents rongeurs piégés en Martinique. Cette étude a permis de mettre en évidence plusieurs mutations dans le gène vkorc1 des rats et des souris. Parmi les souris séquencées, 5 mutations ont été trouvées seules ou en combinaison. Toutes ces mutations étaient des mutations faux-sens A26T, A48T, R61L, Y139C et S149N. La résistance de cible des souris domestiques aux AVK de première génération et même aux AVK de deuxième génération a été clairement démontrée en Martinique surtout avec la détection de la mutation Y139C à une fréquence allélique de 40% et de la double mutation A26T / Y139C à une fréquence allélique de 0,9%. Chez le rat noir, le rongeur le plus répandu en Martinique, 3 nouvelles mutations codantes ont été détectées, les mutations H68N, A115T et S149N associées à une résistance modérée aux AVK de première génération. Parmi les Rattus norvegicus séquencés, une seule mutation silencieuse H68H a été détectée avec une fréquence allélique de 93,3%. Dans un second temps, dans le cadre d’une étude sur le campagnol, nous avons détecté 2 mutations faux-sens (G71R et S149I mutation) avec une très faible prévalence aux seins des populations. Nous avons caractérisé les conséquences catalytiques de ces deux mutations trouvées. Les mutations semblent n’avoir aucun effet sur la résistance aux AVK. Ensuite, nous nous sommes intéressés à la vitamine K chez les campagnols. Le régime alimentaire de ces herbivores est très riche en vitamine K (l’antidote naturel des troubles de coagulation liés à l’utilisation des AVK) et pourrait être responsable de la résistance des campagnols. Ainsi, les campagnols ont montré des concentrations en vitamine K beaucoup plus grandes que celles trouvées chez le rat. En outre, les concentrations de vitamine K chez les campagnols varient avec les saisons, elles sont plus abondantes en été qu’en hiver. Cette variation est liée à la disponibilité de la nourriture qui dépend à son tour des saisons. Il s’agit donc d’une résistance alimentaire. Enfin, nous avons étudié un troisième problème lié aux pullulations des campagnols en évaluant la capacité des campagnols terrestres à participer à la transmission et au maintien du cycle du pathogène dans l’environnement. Ainsi, nous avons cherché à identifier le portage des leptospires par les campagnols dans deux régions de la France touchées par les pullulations (l’Auvergne et la Franche comté). Nous avons identifié la présence, avec une faible prévalence, de L. borgpetersenii et L. kirschneri dans le Cantal et L. borgpetersenii, L. interrogans et L. kirschneri dans le Puy De Dôme. Alors que seule L. kirschneri a été trouvée en Franche comté avec une prévalence beaucoup plus alarmante qu’en Auvergne.
... Vitamin K deficiency impacts neuromuscular and vascular function, thus affecting the physical functioning. Vitamin K has a function in promoting vascular smooth muscle differentiation [34]. As disabilities in patients are directly related to muscle strength and physical performance, therefore it is crucial to focus on muscle strength and performance rather than muscle mass [35]. ...
Full-text available
Menaquinone-7 plays a significant role in cardiovascular and bone health. In recent times there is a growing interest in understanding the role of Menaquinone-7 in health and diseases. Several population-based studies have reported specific health effects of the long-chain menaquinones, notably MK-7, MK-8, and MK-9. There are several epidemiological studies, clinical trials, along with in vivo and in vitro studies confirming the role of Menaquinone-7 in health and diseases. More recently, research group at Synergia Life Sciences has discovered a wider role for Menaquinone-7 in energy homeostasis (VO2max), peripheral neuropathy, muscle cramps and mitochondrial respiration not only through improvement of the electron transport but also the perfusion improving oxygen availability. In the current chapter, the authors have discussed the wider physiological role of Menaquinone-7 highlighting the recent research with Menaquinone-7 in the areas of Muscle and Nerve Health.
... (55) Alternatively, vitamin K promotes vascular smooth muscle differentiation, which may be associated with a better perfusion of muscle tissue. (40) In a cross-sectional study including 1,089 community-dwelling older adults, (33) higher plasma phylloquinone was significantly associated with better physical function, as indicated by short physical performance battery No data of PK and MK intake in all subjects. ...
Full-text available
The most fundamental function of vitamin K is to activate the blood coagulation factors in the liver. Despite the recent recognition of its extra-hepatic actions, the current Dietary Reference Intakes for vitamin K is based on the amount necessary for maintaining the normal blood coagulation in many countries. To define the Dietary Reference Intake for vitamin K, appropriate biomarkers well-reflecting the vitamin K status are essential. Unfortunately, however, no markers are currently available with properties enabling us to properly define the vitamin K status; i.g., no interference by other factors and the presence of widely approved cut-off values. Thus, Adequate Intake is determined, which is an index based on the representative dietary intake data from healthy individuals. Recently, epidemiological studies have been reported regarding the relationship between vitamin K and noncommunicable diseases including osteoporotic fracture. Furthermore, studies focusing on the relationship between vitamin K intake and metabolic syndrome, physical function, depression, cognition, and all-cause mortality have become available, although limited in number. This review summarizes the recent findings in favor of the novel functions of vitamin K. More epidemiological studies are needed to define the appropriate vitamin K intake value based on the prevention of various disorders.
... [3][4][5] Carboxylation is essential for the biological function of VKD proteins that control blood coagulation, vascular calcification, bone metabolism, signal transduction, and cancer cell proliferation. [6][7][8][9][10] VKD carboxylation was originally observed in clotting factors, and carboxylation of coagulation factors has been extensively studied. [11][12][13] Defects in VKD carboxylation are mainly linked to bleeding disorders, known as combined vitamin K-dependent coagulation factors deficiency (VKCFD). ...
Gamma-glutamyl carboxylase (GGCX) is an integral membrane protein that catalyzes posttranslational carboxylation of a number of vitamin K-dependent (VKD) proteins involved in a wide variety of physiological processes, including blood coagulation, vascular calcification, and bone metabolism. Naturally occurring GGCX mutations are associated with multiple distinct clinical phenotypes. However, the genotype-phenotype correlation of GGCX remains elusive. Here, we systematically examined the effect of all naturally occurring GGCX mutations on the carboxylation of three structure-function distinct VKD proteins in a cellular environment. GGCX mutations were transiently introduced into GGCX-deficient human embryonic kidney 293 cells stably expressing chimeric coagulation factor, matrix Gla protein (MGP), or osteocalcin as VKD reporter-proteins, then the carboxylation efficiency of these reporter-proteins were evaluated. Our results show that GGCX mutations differentially affect the carboxylation of these reporter-proteins and the efficiency of using vitamin K as a cofactor. Carboxylation of these reporter-proteins by a C-terminal truncation mutation (R704X) implies that GGCX's C-terminus plays a critical role in the binding of osteocalcin, but not in the binding of coagulation factors and MGP. This has been confirmed by probing the protein-protein interaction between GGCX and its protein substrates in live cells using bimolecular fluorescence complementation and chemical cross-linking assays. Additionally, using a minigene splicing assay, we demonstrated that several GGCX missense mutations affect GGCX's pre-mRNA splicing rather than altering the corresponding amino acid residues. Results from this study interpreted the correlation of GGCX's genotype and its clinical phenotypes, and clarified why vitamin K administration rectified bleeding disorders but not non-bleeding disorders.
... This is because the body is implementing a recycling of vitamin K epoxide, so that it can again interact with GGCX. Thus, a molecule of vitamin K can provide about 500 carboxylation reactions (Chatrou et al., 2011). ...
Full-text available
The present study aims to compare in vitro the pharmacodynamics characteristics between diastereoisomers of each of the three SGARs, bromadiolone, difenacoum, and difethialone toward VKORC1 of field voles (Microtus arvalis) and European water voles (Arvicola terrrestris) expressed in a yeast Pichia pastoris. Ki values were obtained from at least from two separate experiments
... L'apport alimentaire en vitamine K étant limité, l'organisme a mis en place un système de recyclage efficace permettant le maintien d'une quantité suffisante pour assurer les fonctions biologiques. L'efficacité de ce cycle est telle qu'on évalue à environ 500 fois la capacité de régénération d'une molécule de vitamine K avant son élimination (Chatrou et al., 2011). 5/ Sous sa forme KH2 la vitamine K va pouvoir être réutilisée à l'étape 1 pour l'activation d'une nouvelle PVKD par la GGCX. ...
La vitamine K permet l’activation biologique de 14 protéines identifiées à ce jour et qualifiées de vitamines K dépendantes (PVKD). Un recyclage très efficient de la vitamine K par l’enzyme VKORC1 permet d’en limiter grandement les besoins nutritionnels. Chez certaines personnes, une subcarence pourrait exister et contribuer au développement de calcifications de la média des parois vasculaires et augmenter ainsi le facteur de risque cardio-vasculaire. Ce type de calcification est retrouvé chez les personnes hémodialysées, présentant un diabète ou tout simplement au cours du vieillissement. Le lien entre cette subcarence et les calcifications vasculaires est difficilement étudiable chez l’Homme du fait d’une évolution lente et d’effets pouvant être masqués par d’autres facteurs environnementaux. Pour pouvoir étudier directement ce phénomène, un modèle murin a été développé et caractérisé. Les rats développés dans de ce modèle ont des besoins fortement augmentés en vitamine K du fait d’un recyclage fortement altéré. Après 12 semaines d’administration déficient en vitamine K, les rats mâles présentaient des calcifications vasculaires médiales importantes de l’aorte, des poumons, des testicules et du cœur. La coagulation est maintenue bien qu’une diminution de l’activité des facteurs de la coagulation vitamine K dépendants soit observée. La matrix-gla-protéine tissulaire ainsi que l’ostéocalcine plasmatique sont retrouvées accumulées sous leur forme inactive. Dans le même temps, la concentration en vitamine K tissulaire est très fortement diminuée. Ces résultats ne sont pas observés chez les rats non mutés subcarencés ni chez les rats mutés non-subcarencé. Ce travail permet pour la première fois de montrer l’existence d’un lien fort entre subcarence en vitamine K et calcifications vasculaires. Ce modèle de subcarence pourrait servir à une meilleure compréhension du rôle des différentes PVKD extra-hépatiques. Il constitue également un modèle de choix pour l’étude des calcifications médiales et de leurs évolutions
... The latter is then re-reduced first into vitamin K quinone and then into vitamin K hydroquinone by the vitamin K epoxide reductase (VKORC1) enzyme to be reusable for a new activation of VKDP. It is estimated that one molecule of vitamin K can be used up to 500 times by GGCX [13]. Inhibition of the VKORC1 by vitamin K antagonists such as warfarin leads to inhibition of recycling, which results in a hemorrhage due to a deficiency of activated clotting factors II, VII, IX, and X. ...
Full-text available
Vitamin K is crucial for many physiological processes such as coagulation, energy metabolism, and arterial calcification prevention due to its involvement in the activation of several vitamin K-dependent proteins. During this activation, vitamin K is converted into vitamin K epoxide, which must be re-reduced by the VKORC1 enzyme. Various VKORC1 mutations have been described in humans. While these mutations have been widely associated with anticoagulant resistance, their association with a modification of vitamin K status due to a modification of the enzyme efficiency has never been considered. Using animal models with different Vkorc1 mutations receiving a standard diet or a menadione-deficient diet, we investigated this association by measuring different markers of the vitamin K status. Each mutation dramatically affected vitamin K recycling efficiency. This decrease in recycling was associated with a significant alteration of the vitamin K status, even when animals were fed a menadione-enriched diet suggesting a loss of vitamin K from the cycle due to the presence of the Vkorc1 mutation. This change in vitamin K status resulted in clinical modifications in mutated rats only when animals receive a limited vitamin K intake totally consistent with the capacity of each strain to recycle vitamin K.
... Vitamin K plays an important role in the regulation of proteins associated with the inhibition of cardiovascular disease-related complications [45]. It acts as a cofactor for the enzyme γ-glutamyl carboxylase in the post-translational conversion of glutamic acid to γ-carboxyglutamic acid (Gla) residues. ...
... Vitamin K comprises a group of fat-soluble vitamins that act as co-factor for γ-glutamyl carboxylase, which activates several vitamin K-dependent proteins (VKDPs). VKDPs play a major role in hemostasis and bone health, but also in the progression of vascular calcification (VC), which is strongly associated with cardiovascular risk [3] and CKD mortality [4,5]. Several VKDPs also have important roles as ligands in apoptotic pathways with a significant implication in cancer therapy and disease progression [6][7][8]. ...
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Patients with chronic kidney disease (CKD) have an increased risk of developing vascular calcifications, as well as bone dynamics impairment, leading to a poor quality of life and increased mortality. Certain vitamin K dependent proteins (VKDPs) act mainly as calcification inhibitors, but their involvement in the onset and progression of CKD are not completely elucidated. This review is an update of the current state of knowledge about the relationship between CKD and four extrahepatic VKDPs: matrix Gla protein, osteocalcin, growth-arrest specific protein 6 and Gla-rich protein. Based on published literature in the last ten years, the purpose of this review is to address fundamental aspects about the link between CKD and circulating VKDPs levels as well as to raise new topics about how the interplay between molecular weight and charge could influence the modifications of circulating VKDPs at the glomerular level, or whether distinct renal etiologies have effect on VKDPs. This review is the output of a systematic literature search and may open future research avenues in this niche domain.
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Zusammenfassung Eine tägliche Vitamin-D-Supplementierung für Säuglinge bis zum zweiten erlebten Frühsommer zur Prävention der Rachitis und die Gabe von Vitamin K 1 bei Neugeborenen zur Prävention von Vitamin-K-Mangel-Blutungen sind empfohlen. Seit einiger Zeit sind in Österreich Kombinationsprodukte der beiden fettlöslichen Vitamine D 3 und K 2 auf dem Markt erhältlich, die mit gesundheitsfördernden Effekten wie verbesserter Knochenmineralisation und Schutz vor vaskulärer Kalkeinlagerung beworben werden. Die Wirkung einer kombinierten Supplementierung von Vitamin D und Vitamin K 2 bei Kindern ist aus physiologischer Sicht gesehen zwar potenziell sinnvoll, um Effekt, Risiken oder unerwünschte Nebenwirkungen zu evaluieren. Zuvor werden aber Dosisfindungs- und Sicherheitsstudien für die kombinierte Verabreichung benötigt. Insbesondere sind randomisierte kontrollierte Studien bei Risikokindern und Frühgeborenen notwendig. Solange diese Daten nicht vorliegen, erscheint die Gabe von Vitamin D in Kombination mit Vitamin K 2 im Kindesalter nicht gerechtfertigt und kann daher auch nicht empfohlen werden.
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An unusual tryptic peptide isolated from the protein, prothrombin, has recently been shown to contain at least a portion of the vitamin K-dependent region of this protein (Nelsestuen, G. L., and Suttie, J. W. (1973) Proc. Nat. Acad. Sci. U. S. A. 70, 3366–3370). A modified glutamylserine dipeptide has been isolated from proteolytic digests of this vitamin K-dependent peptide and has been characterized. It has been unambiguously demonstrated that the glutamic acid of the glutamylserine has been substituted by carboxylation of the γ carbon atom so that the dipeptide structure is: [see PDF for equation] Presumably, the incorporation of the second γ-carboxyl group is dependent on vitamin K.
A case of haemorrhagic disease of the newborn was reported in order to emphasize the importance of the administration of phytonadione at birth. Since the administration of vitamin K is not considered as a routine prophylaxis, several cases of haemorrhagic disease of the newborn were observed.
Acute myocardial infarction (AMI) is a common and lethal heart disease, and the recruitment of fibroblastic cells to the infarct region is essential for the cardiac healing process. Although stiffness of the extracellular matrix in the infarct myocardium is associated with cardiac healing, the molecular mechanism of cardiac healing is not fully understood. We show that periostin, which is a matricellular protein, is important for the cardiac healing process after AMI. The expression of periostin protein was abundant in the infarct border of human and mouse hearts with AMI. We generated periostin(-/-) mice and found no morphologically abnormal cardiomyocyte phenotypes; however, after AMI, cardiac healing was impaired in these mice, resulting in cardiac rupture as a consequence of reduced myocardial stiffness caused by a reduced number of alpha smooth muscle actin-positive cells, impaired collagen fibril formation, and decreased phosphorylation of FAK. These phenotypes were rescued by gene transfer of a spliced form of periostin. Moreover, the inhibition of FAK or alpha v-integrin, which blocked the periostin-promoted cell migration, revealed that alpha v-integrin, FAK, and Akt are involved in periostin signaling. Our novel findings show the effects of periostin on recruitment of activated fibroblasts through FAK-integrin signaling and on their collagen fibril formation specific to healing after AMI.
Background Electron-beam CT (EBCT) quantification of coronary artery calcification (CAC) allows noninvasive assessment of coronary atherosclerosis. We undertook a follow-up study to determine whether CAC extent, measured at the time of angiography by EBCT, predicted future hard cardiac events, comprising cardiac death and nonfatal myocardial infarction (MI). We also assessed the potential of selected coronary artery disease (CAD) risk factors, prior CAD event history (MI or revascularization), and angiographic findings (number of diseased vessels and overall disease burden) to predict subsequent hard events. Methods and Results Two hundred eighty-eight patients who underwent contemporaneous coronary angiography and EBCT scanning were contacted after a mean of 6.9 years. Vital status and history of MI during follow-up were determined. Cox proportional hazards models were used to compare the predictive ability of CAC extent with selected CAD risk factors, CAD event history, and angiographic findings. Median CAC score was 160 (range 0 to 7633). The 22 patients who experienced hard events during follow-up were older and had more extensive CAC and angiographic disease ( P <0.05). Only 1 of 87 patients with CAC score <20 experienced a subsequent hard event during follow-up. Event-free survival was significantly higher for patients with CAC scores <100 than for those with scores ≥100 (relative risk 3.20; 95% CI 1.17 to 8.71). When a stepwise multivariable model was used, only age and CAC extent predicted hard events (risk ratios 1.72 and 1.88, respectively; P <0.05). Conclusions In patients undergoing angiography, CAC extent on EBCT is highly predictive of future hard cardiac events and adds valuable prognostic information. Received January 4, 2001; revision received April 20, 2001; accepted April 20, 2001.
1. The hemorrahgic agent is spoiled sweet clover hay (Melilotus alaba) has been isolatd in a pure state, m. p. 288-289°, from experimentally produced spoiled hays as well as form hyas that killed cattle in argicultural practice. 2. This substance has the empirical formula C12H12O6. It is optically inactive. There are two acidic hydroxyls in the molecule. The acidity of the pure substance falls between that of the phenols and the carboxylic acids. A crystalline dimethyl other C12H12O4(OCH2)2 with a melting point fo 168-170° (physiologically inactive) has been prepared by methylation with diazomethane. 3. In the pure state the substance has a low soulubility in the ordinary organic solvents. It is insoluble in acid media. Basic solvenets. and dilute alkali effect solution radily (salt formation). 4. The substance could not be characterized or identified on the basis of its behavior towards the usual identification reagents. Its occurrence in nature has not previosly been reproted. 5. 1.5 mg. of hte crystalline hemorrhagic agent cause approximately the same reduciton in the prothrombin level or activity of the plasma of standardized susceptible reabbits in 40 hours as 50 gm. of the standard spoiled hay sample. 6. Spoiled sweet cover hays produced experimentally from Melilotus alba and those relized in agricultural parctice contain approximately 0.003 per cent of the hemorrhagic agent on the dry substance basis. The over-all yield of the subsance in a fractionation scheme involving sixteen steps approximates 86 per cent of the quantity present.
Proinflammatory cytokines such as tumor necrosis factor- (TNF-) are critically involved in activation of the coagulation system in sepsis, leading to disseminated intravascular coagulation (DIC). Natural anticoagulants such as antithrombin (AT) and activated protein C (APC) regulate the coagulation system by inhibiting thrombin generation. In addition to these anticoagulant effects, both AT and APC have been shown to attenuate inflammatory responses induced by various noxious stimuli in rats such as lipopolysaccharide (LPS) challenge. AT promotes the endothelial release of prostacyclin, a potent anti-inflammatory prostaglandin that inhibits the monocytic production of TNF-, by interacting with cell-surface heparin-like substances. APC directly inhibits the production of TNF- by inhibiting the activation of both nuclear factor κB (NFκB) and activator protein-1 in monocytes stimulated with LPS. Thrombomodulin, an endothelial membranous integral protein that binds thrombin, exerts anti-inflammatory effects by generating APC. Furthermore, tissue factor pathway inhibitor, a natural anticoagulant for the extrinsic pathway of the coagulation system, also attenuates LPS-induced inflammatory responses in rats by inhibiting TNF- production by monocytes. These findings strongly suggest that natural anticoagulants could regulate inflammatory responses as well as the coagulation system in rats by inhibiting the monocytic production of TNF-. Such anti-inflammatory properties of natural anticoagulants are potentially important for their replacement in patients with sepsis who frequently develop DIC and organ failure as inflammatory responses.