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Menaquinone-7 Supplementation Improves Arterial Stiffness in Healthy Postmenopausal Women: Double-Blind Randomised Clinical Trial

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Menaquinone-7 Supplementation Improves Arterial Stiffness in Healthy Postmenopausal Women: Double-Blind Randomised Clinical Trial Cardiovascular Biology and Cell Signalling 1 Menaquinone-7 supplementation improves arterial stiffness in healthy postmenopausal women: double-blind randomised clinical trial Marjo H. J. Knapen1; Lavienja A. J. L. M. Braam1; Nadja E. Drummen1; Otto Bekers2; Arnold P. G. Hoeks3; Cees Vermeer1 1VitaK & Cardiovascular Research Institute (CARIM), Maastricht University, The Netherlands; 2Central Diagnostic Laboratory, University Hospital Maastricht, The Netherlands; 3Biomedical Engineering, Maastricht University, The Netherlands Summary At baseline dp-ucMGP was associated with IMT, Diameter, cfPWV and Observational data suggest a link between menaquinone (MK, vit- with the mean z-scores of acute phase markers (APMscore) and of amin K2) intake and cardiovascular (CV) health. However, MK inter- markers for endothelial dysfunction (EDFscore). After three year MK-7 vention trials with vascular endpoints are lacking. We investigated supplementation cfPWV and the Stiffness Index β significantly de- long-term effects of MK-7 (180 µg MenaQ7/day) supplementation on creased in the total group, whereas distension, compliance, distensi- arterial stiffness in a double-blind, placebo-controlled trial. Healthy bility, Young’s Modulus, and the local carotid PWV (cPWV) improved in postmenopausal women (n=244) received either placebo (n=124) or women having a baseline Stiffness Index β above the median of 10.8. MK-7 (n=120) for three years. Indices of local carotid stiffness (intima- MK-7 decreased dp-ucMGP by 50 % compared to placebo, but did not media thickness IMT, Diameter end-diastole and Distension) were influence the markers for acute phase and endothelial dysfunction. In measured by echotracking. Regional aortic stiffness (carotid-femoral conclusion, long-term use of MK-7 supplements improves arterial and carotid-radial Pulse Wave Velocity, cfPWV and crPWV, respect- stiffness in healthy postmenopausal women, especially in women hav- ively) was measured using mechanotransducers. Circulating desp- ing a high arterial stiffness. hospho-uncarboxylated matrix Gla-protein (dp-ucMGP) as well as acute phase markers Interleukin-6 (IL-6), high-sensitive C-reactive Keywords protein (hsCRP), tumour necrosis factor-α (TNF-α) and markers for en- Arterial stiffness, carotid intima-media thickness, menaquinone-7, dothelial dysfunction Vascular Cell Adhesion Molecule (VCAM), E-se- pulse wave velocity, Stiffness Index β lectin, and Advanced Glycation Endproducts (AGEs) were measured. Correspondence to: Received: August 14, 2014 Cees Vermeer, PhD Accepted after major revision: December 31, 2014 VitaK, Maastricht University, Biopartner Center Maastricht Epub ahead of print: February 19, 2015 Oxfordlaan 70, 6229 EV Maastricht, The Netherlands http://dx.doi.org/10.1160/TH14-08-0675 Tel: +31 43 388 5865, fax: +31 43 388 5889 Thromb Haemost 2015; 113: E-mail: [email protected] Financial support: This work was supported by Nattopharma (Høvik, Norway). Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the au- thors, and do not necessarily reflect the view of Nattopharma. Introduction elastic properties of the arterial wall material (Young’s Modulus) and the Stiffness Index β (1, 8). Arterial stiffening accompanying In recent years, there has been a strong focus on the role of arterial age and other CV risk factors is caused by various phenomena, in- stiffness in the development of cardiovascular disease (CVD). Ar- cluding breaks in elastin fibres, accumulation of collagen, fibrosis, terial stiffness can be measured directly and non-invasively (1) and medial smooth muscle necrosis, diffusion of macromolecules in has emerged as independent predictor of CV risk (2, 3). Carotid- the arterial wall, inflammation, and calcification (1). Vascular cal- femoral pulse wave velocity (cfPWV) is considered the gold-stan- cification can modify both functional and structural arterial prop- dard measurement of regional arterial stiffness and has been dem- erties, known as arterial remodelling, resulting in increased arter- onstrated in epidemiological studies the predictive value of aortic ial stiffness. stiffness for CV events (1, 4–6). Measurement of local carotid stiff- Observational studies showed lower prevalence of arterial cal- ness may also provide important prognostic information since the cification and coronary heart disease mortality in subjects with the carotid artery is a frequent site of atheroma formation (1). Indices highest intake of menaquinones (MKs, vitamin K2) (9–11). Re- of local carotid stiffness include: carotid pulse wave velocity markably, no effect was seen for phylloquinone (vitamin K1) in- (cPWV) based on carotid diameter and pulse pressure (7), arterial take in these studies. This is in line with an earlier publication in wall thickness (carotid intima-media thickness, IMT), elastic which no relation was found between dietary phylloquinone in- properties of the artery as a whole (Distensibility, Compliance), take and premature coronary calcification (12). Erkkilä et al. found © Schattauer 2015 Thrombosis and Haemostasis 113.5/2015 Downloaded from www.thrombosis-online.com on 2015-02-22 | ID: 1001065452 | IP: 192.114.4.135 Note: Uncorrected proof, epub ahead of print online For personal or educational use only. No other uses without permission. All rights reserved. 2 Knapen et al. Menaquinone-7 supplementation and cardiovascular health an association between phylloquinone intake and coronary heart body mass index (BMI) > 30 kg/m2, osteoporotic at baseline disease, but it was lost after adjustment for dietary variables (13, (T-score ≤-2.5 SD), coagulation disorders, chronic diseases, meta- 14). While phylloquinone is primarily found in green leafy veg- bolic bone diseases, gastrointestinal diseases, medication that in- etables, MKs occur, but to a much lower extent, in meat (menaqui- terferes with vitamin K and/or blood coagulation, use of corticos- none-4, MK-4) and fermented foods, like cheeses (MK-8 to teroids, bisphosphonates, or hormone replacement therapy, use of MK-10) in Western diets and natto (MK-7) in Japan (15). Up to supplements containing vitamin K, participation in a clinical study now, only two randomised clinical trials have evaluated the effects three months prior to this study, and soy allergy. Participants were of vitamin K supplements on CV health (16, 17) and showed randomly assigned to receive placebo capsules (n=124) or capsules beneficial effects with high daily dosages (0.5–1 mg) of phylloqui- containing 180 µg MK-7 (MenaQ7, NattoPharma, Høvik, Nor- none. To date, MK intervention trials with CV endpoints are lack- way) (n=120). During the trial were no dietary restrictions. From ing, despite the increasing retail availability of MK-7 supplements. the 244 volunteers who entered the study, 21 discontinued their Its longer half-life and higher efficacy compared with phylloqui- participation and were not available for the follow-up measure- none (18) justify the use of much lower doses of MK-7 in trials stu- ments. Participants came to the research site (VitaK, Maastricht, dying its effects on CV health. The Netherlands) every year for blood sampling and measure- The circulating inactive form of matrix Gla-protein (MGP), i. e. ments of body weight, height, and vascular parameters. Methods desphospho-uncarboxylated MGP (dp-ucMGP), a marker for vas- for blood sampling and anthropometrics were described pre- cular vitamin K status (19), has been associated with arterial cal- viously (37). cification and CV mortality (20–25). Vitamin K mediates the car- This study was conducted according to the guidelines laid boxylation of protein-bound glutamate (Glu) residues into γ-car- down in the Declaration of Helsinki and all procedures involving boxyglutamate (Gla) in MGP resulting in protein functionality, i. e. human subjects were approved by the Medical Ethics Committee local calcification inhibition (15). Substantial amounts of dp- of the Maastricht University (Maastricht, The Netherlands). ucMGP can be found in the circulation of non-supplemented Written informed consent was obtained from all subjects before healthy individuals, indicative of vascular vitamin K insufficiency. entering the study. Trial registration code: clinicaltrials.gov Recent work showed that vitamin K supplementation improves NCT00642551. MGP carboxylation (26–29), but the question remains whether this beneficially affects CV health. Next to its role in carboxylation, Study products vitamin K may influence arterial stiffness by carboxylation-inde- pendent mechanisms. Inflammation plays also a role in arterial re- Capsules containing 180 µg MK-7 (MenaQ7, Nattopharma, modelling (30), possibly by affecting proliferation of vascular Høvik, Norway) and matching placebo capsules (with the same smooth muscle cells, influx of leucocytes, and/or production of composition, except for the active component MK-7) were manu- proinflammatory markers. In vitro studies suggest that vitamin K factured by EuroPharma Alliance (Wroclaw, Poland) for Nattoph- may suppress inflammation by decreasing gene expression of arma (Høvik, Norway). They were delivered directly by Nattoph- proinflammatory markers (31). Consistently, observational data arma to VitaK (Maastricht, The Netherlands). One capsule was showed an inverse association between vitamin K status and taken daily either with breakfast or dinner during a period of 36 proinflammatory markers (26, 32–34). Although phylloquinone months. Based on a 3-fold higher efficacy of MK-7 than
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