Relationships between the expression of adipocytokine genes and the calcification of coronary arteries in patients with coronary artery disease

Dysfunctional changes and remodeling of adipose tissue (АT) are associated with the formation of microcalcifications in the vascular wall. Biologically active substances synthesized by АT (adipocytokines) can act as promoters and inhibitors  of vascular calcification development. The few available experimental and clinical studies do not fully explain the possible mechanisms of these effects.

Aim. To study the relationships between the adipocytokine profiles of adipocytes in epicardial and perivascular AT with the severity of coronary artery calcification in patients with coronary artery disease (CAD).

Material and Methods. A total of 125 patients with CAD aged 59 (53; 66) years were examined. The isolated adipocytes of subcutaneous adipose tissue (SAT), epicardial adipose tissue (EAT), and perivascular adipose tissue (PVAT), obtained during coronary artery bypass grafting, were used to determine gene expression and secretion of adipocytokines (adiponectin, leptin, and IL-6). Expression of adipocytokine genes was assessed using quantitative PCR with detection of products in real time (real-time qPCR); the concentration of adipocytokines in the culture medium was determined by enzyme-linked immunosorbent assay using R&D Systems kits (Canada). Coronary artery (CA) calcification degree was assessed by multislice spiral computed tomography (MSCT) method. The calcium index of CA was determined by the Agatston method using the Syngo Calcium Scoring software package (Siemens AG Medical Solution, Germany).

Results. Massive coronary calcification (CC) had the highest prevalence (58.8%) in patients with CAD. The highest level of expression of the ADIPOQ gene in all types of fat stores was observed in patients with moderate/medium CС compared to those with massive CС; the maximum expression of ADIPOQ was observed in the culture of PVAT adipocytes. Expression of the LEP and IL6 genes in massive CC was higher, with the maximum values in the culture of EAT adipocytes relative to SAT and PVAT adipocytes. Decreases in the levels of ADIPOQ mRNA and its secretion, increases in the levels of mRNA of LEP and IL6 and their secretion in adipocytes of the EAT and PVAT were associated with the development of СС in patients with CAD.

Conclusion. Proinflammatory adipokines produced by adipocytes of patients with CAD during hypoxia induced vascular calcification by stimulating oxidative stress, osteoblast differentiation, apoptosis, and proliferation of smooth muscle cells. Endothelial cells, when stimulated with proinflammatory adipocytokines, tended to transform into osteoblasts, which further aggravated the degree of vascular inflammation and calcification.

1. Gruzdeva O.V., Akbasheva O.E., Dyleva Yu.A., Antonova L.V., Matveeva V.G., Uchasova E.G. et al. Adipokine and cytokine profiles of epicardial and subcutaneous adipose tissue in patients with coronary artery disease. Bull. Exp. Biol. Med. 2017;163(5):608–611 (In Russ.). DOI: 10.1007/s10517-017-3860-5.

2. Dyleva Yu.A., Gruzdeva O.V., Belik E.V., Akbasheva O.E., Uchasova E.G., Borodkina D.A. et al. Gene expression and adiponectin content in adipose tissue in patients with coronary artery disease. Biomed. Khim. 2019;65(3):239–244 (In Russ.). DOI: 10.18097/pbmc20196503239.

3. Gruzdeva O., Uchasova E., Dyleva Yu., Borodkina D., Akbasheva O., Antonova L. et al. Adipocytes directly affect coronary artery disease pathogenesis via induction of adipokine and cytokine imbalances. Front. Immunol. 2019;10:2163. DOI: 10.3389/fimmu.2019.02163.

4. Son B.-K., Akishita M., Iijima K., Kozaki K., Maemura K., Eto M. et al. Adiponectin antagonizes stimulatory effect of tumor necrosis factor-α on vascular smooth muscle cell calcification: Regulation of growth arrest-specific gene 6-mediated survival pathway by adenosine 5′-monophosphate-activated protein kinase. Endocrinology. 2008;149(4):1646– 1653. DOI: 10.1210/en.2007-1021.

5. Luo X.H., Zhao L.L., Yuan L.Q., Wang M., Xie H., Liao E.Y. Development of arterial calcification in adiponectin-deficient mice: Adiponectin regulates arterial calcification. J. Bone Miner. Res. 2009;24(8):1461–1468. DOI: 10.1359/jbmr.090227.

6. Fukuyo S., Yamaoka K., Sonomoto K., Oshita K., Okada Y., Saito K. et al. IL-6-accelerated calcification by induction of ROR2 in human adipose tissue-derived mesenchymal stem cells is STAT3 dependent. Rheumatology. 2014;53(7):1282–1290. DOI: 10.1093/rheumatology/ket496.

7. Larsen B.A., Laughlin G.A., Cummins K., Barrett-Connor E., Wasse C.L. Adipokines and severity and progression of coronary artery calcium: Findings from the Rancho Bernardo Study. Atherosclerosis. 2017;265:1–6. DOI: 10.1016/j.atherosclerosis.2017.07.022.

8. Du B., Ouyang A., Eng J.S., Fleenor B.S. Aortic perivascular adipose-derived interleukin-6 contributes to arterial stiffness in low-density lipoprotein receptor deficient mice. Am. J. Physiol. Heart Circulatory Physiol. 2015;308(11):H1382–H1390. DOI: 10.1152/ajpheart.00712.2014.

9. . Yao Y., Watson A.D., Ji S., Bostrom K.I. Heat shock protein 70 enhances vascular bone morphogenetic protein-4 signaling by binding matrix Gla protein. Circ. Res. 2009;105(6):575–584. DOI: 10.1161/circresaha.109.202333.

10. Reilly M.P., Iqbal N., Schutta M., Wolfe M.L., Scally M., Localio A.R. et al. Plasma leptin levels are associated with coronary atherosclerosis in type 2 diabetes. J. Clin. Endocrinol. Metab. 2004;89(8):3872–3878. DOI: 10.1210/jc.2003-031676.

11. Qasim A., Mehta N.N., Tadesse M.G., Wolfe M.L., Rhodes T., Girman C. et al. Adipokines, insulin resistance and coronary artery calcification. J. Am. Coll. Cardiol. 2008;52(3): 231–236. DOI: 10.1016/j.jacc.2008.04.016.

12. Parhami F., Tintut Y., Ballard A., Fogelman A.M., Demer L.L. Leptin enhances the calcification of vascular cells: artery wall as a target of leptin. Circ. Res. 2001;88(9):954–960. DOI: 10.1161/hh0901.090975.

13. Zeadin M., Butcher M., Werstuck G., Khan M., Yee C.K., Shaughnessy S.G. Effect of leptin on vascular calcification in apolipoprotein E-deficient mice. Arterioscler. Thromb. Vasc. Biol. 2009;29(12):2069–2075. DOI: 10.1161/atvbaha.109.195255.

14. Dong F., Zhang X., Ren J. Leptin regulates cardiomyocyte con tractile function through endothelin-1 receptor-NADPH oxidase pathway. Hypertension. 2006;47(2):222–229. DOI: 10.1161/01.hyp.0000198555.51645.f1.

15. Schroeter M.R., Stein S., Heida N.-M., Leifheit-Nestler M., Cheng I.-F., Gogiraju R. et al. Leptin promotes the mobilization of vascular progenitor cells and neovascularization by NOX2-mediated activation of MMP9. Cardiovasc. Res. 2012;93(1):170–180. DOI: 10.1093/cvr/cvr275.

16. Zeidan A., Purdham D.M., Rajapurohitam V., Javadov S., Chakrabarti S., Karmazyn M. Leptin induces vascular smooth muscle cell hypertrophy through angiotensin IIand endothelin-1-dependent mechanisms and mediates stretchinduced hypertrophy. J. Pharmacol. Exp. Ther. 2005;315(3):1075–1084. DOI: 10.1124/jpet.105.091561.

17. Fantuzzi G., Faggioni R. Leptin in the regulation of immunity, inflammation, and hematopoiesis. J. Leukoc. Biol. 2000;68(4):437–446.

18. Bastard J.P., Maachi M., Lagathu C., Kim M.J., Caron M., Vidal H. et al. Recent advances in the relationship between obesity, inflammation, and insulin resistance. Eur. Cytokine Netw. 2006;17(1):4–12.

19. Vaughan T., Li L. Molecular mechanism underlying the inflammatory complication of leptin in macrophages. Mol. Immunol. 2010;47(15):2515– 2518. DOI: 10.1016/j.molimm.2010.06.006.

20. Byon C.H., Javed A., Dai Q., Kappes J.C., Clemens T.L., Darley-Usmar V.M. et al. Oxidative stress induces vascular calcification through modulation of the osteogenic transcription factor Runx2 by AKT signaling. J. Biol. Chem. 2008;283(22):15319–15327. DOI: 10.1074/jbc. m800021200.

21. Szasz T., Bomfim G.F., Webb R.C. The influence of perivascular adipose tissue on vascular homeostasis. Vascular Health and Risk Management. 2013;9:105–116. DOI: 10.2147/vhrm.s33760.

22. Fern´andez-Alfonso M.S., Gil-Ortega M., García-Prieto C.F., Aranguez I., RuizGayo M., Somoza B. Mechanisms of perivascular adipose tissue dysfunction in obesity. Int. J. Endocrinol. 2013;2013:402053. DOI: 10.1155/2013/402053.