Menaquinone‑4 modulates the expression levels of calcification‑associated factors to inhibit calcification of rat aortic vascular smooth muscle cells in a dose‑dependent manner

Cui,Liwen; Xu,Jinsheng; Zhang,Junxia; Zhang,Muqing; Zhang,Shenglei; Bai,Yaling

doi:10.3892/etm.2018.6535

Menaquinone‑4 modulates the expression levels of calcification‑associated factors to inhibit calcification of rat aortic vascular smooth muscle cells in a dose‑dependent manner

Authors:
- Liwen Cui
- Jinsheng Xu
- Junxia Zhang
- Muqing Zhang
- Shenglei Zhang
- Yaling Bai
View Affiliations

Affiliations: Department of Nephrology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China
Published online on: July 27, 2018 https://doi.org/10.3892/etm.2018.6535
Pages: 3172-3178

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Vascular calciﬁcation (VC) caused by chronic kidney disease (CKD)‑mineral and bone disorder is a common complication of CKD. Recent studies have demonstrated that menaquinone‑4 (MK‑4) is negativly associated with VC in patients with CKD. Furthermore, we have previously shown that runt‑related transcription factor 2 (Runx2) is important in the phenotypic transformation process of rat vascular smooth muscle cells (VSMCs), which is the key step for the development of VC. The present study investigated the influence of MK‑4 on the phenotypic transformation process of rat VSMCs in order to illustrate its role in the process of VC. Calcification assays were perfomed to access the calcified degree of rat VSMCs. Additionally, the genes and proteins related to phenotypic transformation were measured by reverse transcription‑polymerase chain reaction and western blotting methods. It was revealed that calcium deposition in the cells was evidently increased with an addition of β‑glycerophosphate (β‑GP) and could be completely prevented by co‑incubation with MK‑4 in a dose‑dependent manner. Furthermore, the expression of Runx2 in the β‑GP‑induced VSMCs was inhibited by MK‑4. It was also revealed that the expression of SMAD1 and bone morphogenetic protein (BMP)‑2 were decreased in the β‑GP‑induced VSMCs treated with MK‑4 in a dose‑dependent manner; however, the expression of SMAD7 was increased in the β‑GP‑induced VSMCs treated with MK‑4 in a dose‑dependent manner. These observations suggest that MK‑4 reduces mineralization by regulating the BMP‑2 signaling pathway in order to attenuate the expression of Runx2.

View Figures

View References

1	Tonelli M, Wiebe N, Culleton B, House A, Rabbat C, Fok M, McAlister F and Garg AX: Chronic kidney disease and mortality risk: A systematic review. J Am Soc Nephrol. 17:2034–2047. 2006. View Article : Google Scholar : PubMed/NCBI
2	Tentori F, Blayney MJ, Albert JM, Gillespie BW, Kerr PG, Bommer J, Young EW, Akizawa T, Akiba T, Pisoni RL, et al: Mortality risk for dialysis patients with different levels of serum calcium, phosphorus, and PTH: The dialysis outcomes and practice patterns study (DOPPS). Am J Kidney Dis. 52:519–530. 2008. View Article : Google Scholar : PubMed/NCBI
3	Bolasco P: Effects of the use of Non-calcium phosphate binders in the control and outcome of vascular calcifications: A review of clinical trials on CKD patients. Int J Nephrol. 2011:7584502011. View Article : Google Scholar : PubMed/NCBI
4	Adragão T: Evaluation of vascular calcifications in CKD patients. Int J Artif Organs. 32:81–86. 2009. View Article : Google Scholar : PubMed/NCBI
5	Giachelli CM: The emerging role of phosphate in vascular calcification. Kidney Int. 75:890–897. 2009. View Article : Google Scholar : PubMed/NCBI
6	Lau WL and Ix JH: Clinical detection, risk factors, and cardiovascular consequences of medial arterial calcification: A pattern of vascular injury associated with aberrant mineral metabolism. Semin Nephrol. 33:93–105. 2013. View Article : Google Scholar : PubMed/NCBI
7	Olauson H and Larsson TE: FGF23 and Klotho in chronic kidney disease. Curr Opin Nephrol Hypertens. 22:397–404. 2013. View Article : Google Scholar : PubMed/NCBI
8	Li X, Yang HY and Giachelli CM: Role of the sodium-dependent phosphate cotransporter, Pit-1, in vascular smooth muscle cell calcification. Circ Res. 98:905–912. 2006. View Article : Google Scholar : PubMed/NCBI
9	Olszak IT, Poznansky MC, Evans RH, Olson D, Kos C, Pollak MR, Brown EM and Scadden DT: Extracellular calcium elicits a chemokinetic response from monocytes in vitro and in vivo. J Clin Invest. 105:1299–1305. 2000. View Article : Google Scholar : PubMed/NCBI
10	Steitz SA, Speer MY, Curinga G, Yang HY, Haynes P, Aebersold R, Schinke T, Karsenty G and Giachelli CM: Smooth muscle cell phenotypic transition associated with calcification: Upregulation of Cbfa1 and downregulation of smooth muscle lineage markers. Circ Res. 89:1147–1154. 2001. View Article : Google Scholar : PubMed/NCBI
11	Schurgers LJ, Uitto J and Reutelingsperger CP: Vitamin K-dependent carboxylation of matrix Gla-protein: A crucial switch to control ectopic mineralization. Trends Mol Med. 19:217–226. 2013. View Article : Google Scholar : PubMed/NCBI
12	Wallin R, Schurgers LJ and Loeser RF: Biosynthesis of the vitamin K-dependent matrix Gla protein (MGP) in chondrocytes: A fetuin-MGP protein complex is assembled in vesicles shed from normal but not from osteoarthritic chondrocytes. Osteoarthritis Cartilage. 18:1096–1103. 2010. View Article : Google Scholar : PubMed/NCBI
13	Cranenburg EC, VAN Spaendonck-Zwarts KY, Bonafe L, Crettol Mittaz L, Rödiger LA, Dikkers FG, VAN Essen AJ, Superti-Furga A, Alexandrakis E, Vermeer C, et al: Circulating matrix γ-carboxyglutamate protein (MGP) species are refractory to vitamin K treatment in a new case of Keutel syndrome. J Thromb Haemost. 9:1225–1235. 2011. View Article : Google Scholar : PubMed/NCBI
14	Holden RM, Morton AR, Garland JS, Pavlov A, Day AG and Booth SL: Vitamins K and D status in stages 3–5 chronic kidney disease. Clin J Am Soc Nephrol. 5:590–597. 2010. View Article : Google Scholar : PubMed/NCBI
15	Holden RM, Iliescu E, Morton AR and Booth SL: Vitamin K status of Canadian peritoneal dialysis patients. Perit Dial Int. 28:415–418. 2008.PubMed/NCBI
16	Westenfeld R, Krueger T, Schlieper G, Cranenburg EC, Magdeleyns EJ, Heidenreich S, Holzmann S, Vermeer C, Jahnen-Dechent W, Ketteler M, et al: Effect of vitamin K2 supplementation on functional vitamin K deficiency in hemodialysis patients: A randomized trial. Am J Kidney Dis. 59:186–195. 2012. View Article : Google Scholar : PubMed/NCBI
17	Geleijnse JM, Vermeer C, Grobbee DE, Schurgers LJ, Knapen MH, van der Meer IM, Hofman A and Witteman JC: Dietary intake of menaquinone is associated with a reduced risk of coronary heart disease: The Rotterdam Study. J Nutr. 134:3100–3105. 2004. View Article : Google Scholar : PubMed/NCBI
18	Koitaya N, Sekiguchi M, Tousen Y, Nishide Y, Morita A, Yamauchi J, Gando Y, Miyachi M, Aoki M, Komatsu M, et al: Low-dose vitamin K2 (MK-4) supplementation for 12 months improves bone metabolism and prevents forearm bone loss in postmenopausal Japanese women. J Bone Miner Metab. 32:142–150. 2014. View Article : Google Scholar : PubMed/NCBI
19	Campbell JH and Campbell GR: Culture techniques and their applications to studies of vascular smooth muscle. Clin Sci (Lond). 85:501–513. 1993. View Article : Google Scholar : PubMed/NCBI
20	Chiba Y, Sakai H, Wachi H, Sugitani H, Seyama Y and Misawa M: Upregulation of rhoA mRNA in bronchial smooth muscle of antigen-induced airway hyperresponsive rats. J Smooth Muscle Res. 39:221–228. 2003. View Article : Google Scholar : PubMed/NCBI
21	Zhang S, Xu J, Feng Y, Zhang J, Cui L, Zhang H and Bai Y: Extracellular acidosis suppresses calcification of vascular smooth muscle cells by inhibiting calcium influx via L-type calcium channels. Clin Exp Hypertens. 40:370–377. 2018. View Article : Google Scholar : PubMed/NCBI
22	Xu J, Bai Y, Jin J, Zhang J, Zhang S, Cui L and Zhang H: Magnesium modulates the expression levels of calcification-associated factors to inhibit calcification in a time-dependent manner. Exp Ther Med. 9:1028–1034. 2015. View Article : Google Scholar : PubMed/NCBI
23	Cherbuin N: Higher dietary intakes of potassium, calcium and magnesium are associated with a reduced risk of developing vascular dementia. Evid Based Ment Health. 16:262013. View Article : Google Scholar : PubMed/NCBI
24	Chaabane C, Heizmann CW and Bochaton-Piallat ML: Extracellular S100A4 induces smooth muscle cell phenotypic transition mediated by RAGE. Biochim Biophys Acta. 1853:2144–2157. 2014. View Article : Google Scholar : PubMed/NCBI
25	Spronk HM, Soute BA, Schurgers LJ, Thijssen HH, De Mey JG and Vermeer C: Tissue-specific utilization of menaquinone-4 results in the prevention of arterial calcification in warfarin-treated rats. J Vasc Res. 40:531–537. 2003. View Article : Google Scholar : PubMed/NCBI
26	Wahlqvist ML, Tanaka K and Tzeng BH: Clinical decision-making for vitamin K-1 and K-2 deficiency and coronary artery calcification with warfarin therapy: Are diet, factor Xa inhibitors or both the answer? Asia Pac J Clin Nutr. 22:492–496. 2013.PubMed/NCBI
27	Koitaya N, Ezaki J, Nishimuta M, Yamauchi J, Hashizume E, Morishita K, Miyachi M, Sasaki S and Ishimi Y: Effect of low dose vitamin K2 (MK-4) supplementation on bio-indices in postmenopausal Japanese women. J Nutr Sci Vitaminol (Tokyo). 55:15–21. 2009. View Article : Google Scholar : PubMed/NCBI
28	Kerage D, Brindley DN and Hemmings DG: Review: Novel insights into the regulation of vascular tone by sphingosine 1-phosphate. Placenta. 35 Suppl:S86–S92. 2014. View Article : Google Scholar : PubMed/NCBI
29	Wang W, Li C, Pang L, Shi C, Guo F, Chen A, Cao X and Wan M: Mesenchymal stem cells recruited by active TGFβ contribute to osteogenic vascular calcification. Stem Cells Dev. 23:1392–1404. 2014. View Article : Google Scholar : PubMed/NCBI
30	Danilevicius CF, Lopes JB and Pereira RM: Bone metabolism and vascular calcification. Braz J Med Biol Res. 40:435–442. 2007. View Article : Google Scholar : PubMed/NCBI
31	Lin ME, Chen T, Leaf EM, Speer MY and Giachelli CM: Runx2 expression in smooth muscle cells is required for arterial medial calcification in mice. Am J Pathol. 185:1958–1969. 2015. View Article : Google Scholar : PubMed/NCBI
32	Li M, Wu P, Shao J, Ke Z, Li D and Wu J: Losartan inhibits vascular calcification by suppressing the BMP2 and Runx2 expression in rats in vivo. Cardiovasc Toxicol. 16:172–181. 2016. View Article : Google Scholar : PubMed/NCBI
33	Bai Y, Zhang J, Xu J, Cui L, Zhang H and Zhang S: Alteration of type I collagen in the radial artery of patients with end-stage renal disease. Am J Med Sci. 349:292–297. 2015. View Article : Google Scholar : PubMed/NCBI
34	Lieberman JR, Daluiski A and Einhorn TA: The role of growth factors in the repair of bone. Biology and clinical applications. J Bone Joint Surg Am. 84-A:1032–1044. 2002. View Article : Google Scholar : PubMed/NCBI
35	Sun SX, Guo HH, Zhang J, Yu B, Sun KN and Jin QH: BMP-2 and titanium particles synergistically activate osteoclast formation. Braz J Med Biol Res. 47:461–469. 2014. View Article : Google Scholar : PubMed/NCBI
36	Miyazono K, Kamiya Y and Morikawa M: Bone morphogenetic protein receptors and signal transduction. J Biochem. 147:35–51. 2010. View Article : Google Scholar : PubMed/NCBI
37	Kawabata M, Imamura T and Miyazono K: Signal transduction by bone morphogenetic proteins. Cytokine Growth Factor Rev. 9:49–61. 1998. View Article : Google Scholar : PubMed/NCBI
38	Sato Simoes AY, Bub GL and Campos AH: BMP-2 and −4 produced by vascular smooth muscle cells from atherosclerotic lesions induce monocyte chemotaxis through direct BMPRII activation. Atherosclerosis. 235:45–55. 2014. View Article : Google Scholar : PubMed/NCBI
39	Yao Y, Bennett BJ, Wang X, Rosenfeld ME, Giachelli C, Lusis AJ and Boström KI: Inhibition of bone morphogenetic proteins protects against atherosclerosis and vascular calcification. Circ Res. 107:485–494. 2010. View Article : Google Scholar : PubMed/NCBI
40	Cai J, Pardali E, Sánchez-Duffhues G and Ten Dijke P: BMP signaling in vascular diseases. FEBS Lett. 586:1993–2002. 2012. View Article : Google Scholar : PubMed/NCBI
41	Shao JS, Aly ZA, Lai CF, Cheng SL, Cai J, Huang E, Behrmann A and Towler DA: Vascular Bmp Msx2 Wnt signaling and oxidative stress in arterial calcification. Ann N Y Acad Sci. 1117:40–50. 2007. View Article : Google Scholar : PubMed/NCBI