Effects of metformin on ceramide production in local fat depots in patients with coronary artery disease

Aim. To evaluate the effects of metformin on the expression level of ceramide (Cer) synthesis enzyme genes, and on the total content and spectrum of ceramides in local fat depots in patients with coronary heart disease (CHD).
Materials and methods. The study included 41 patients with CHD, who were scheduled to undergo coronary artery bypass graft. Samples of subcutaneous, epicardial, and perivascular adipose tissue (SAT, EAT, and PVAT) were obtained during surgery, adipose tissue adipocytes were isolated and cultured for 24 hours with the addition of metformin (Sigma-Aldrich, USA) at concentrations of 1 and 10 mmol/l. The expression characteristics of de novo ceramide synthesis enzymes were assessed by quantitative polymerase chain reaction on a ViiA 7 amplifier (Applied Biosystems, USA). The total content and spectrum of Cer were determined by high-performance liquid chromatography with mass spectrometry (HPLC-MS/MS). Statistical processing of the results was performed using the STATISTICA 12 software package.
Results. Metformin at a concentration of 1 mmol/l statistically significantly decreased the gene expression of the enzymes serine palmitoyltransferase-1 (SPTLC1) in SAT, EAT and PVAT, ceramide synthase-2 (CERS2) in EAT and SAT, ceramide synthase-4 (CERS4) in EAT and PVAT, and increased dihydroceramide desaturase (DEGS1) in EAT and SAT. Metformin at a concentration of 10 mmol/l decreased the level of SPTLC1 and CERS2 mRNA in EAT, CERS4 in EAT and PVAT. A significant decrease in the content of Cer d18:1/16:0 in EAT at a metformin concentration of 1 mmol/l, and Cer d18:1/18:0 in EAT and PVAT, Cer d18:1/20:0 in SAT and EAT at concentrations of 1 and 10 mmol/l was revealed.
Conclusion. Thus, the potential ability of metformin to inhibit de novo ceramide synthesis in isolated SAT, EAT and PVAT adipocytes and to reduce the content of ceramides associated with adverse cardiovascular events in fat cells was demonstrated in in vitro experiments.

1. Drzewoski J., Hanefeld M. The current and potential therapeutic use of metformin-the good old drug. Pharmaceuticals (Basel). 2021;14(2):122. DOI: 10.3390/ph14020122.

2. Miklankova D., Markova I., Hüttl M., Zapletalova I., Poruba M., Malinska H. Metformin affects cardiac arachidonic acid metabolism and cardiac lipid metabolite storage in a prediabetic rat model. Int. J. Mol. Sci. 2021;22(14):7680. DOI: 10.3390/ijms22147680.

3. Seifarth C., Schehler B., Schneider H.J. Effectiveness of metformin on weight loss in non-diabetic individuals with obesity. Exp. Clin. Endocrinol. Diabetes. 2013;121(1):27–31. DOI: 10.1055/s-0032-1327734.

4. Halabi A., Sen J., Huynh Q., Marwick T.H. Metformin treatment in heart failure with preserved ejection fraction: a systematic review and meta-regression analysis. Cardiovasc. Diabetol. 2020;19(1):124. DOI: 10.1186/s12933-020-01100-w.

5. Lascu A., Ionică L.N., Merce A.P., Dănilă M.D., Petrescu L., Sturza A. et al. Metformin Acutely Mitigates Oxidative Stress in Human Atrial Tissue: A Pilot Study in Overweight Non-Diabetic Cardiac Patients. Life (Basel). 2022;12(12):2058. DOI: 10.3390/life12122058.

6. Bai B., Chen H. Metformin: a novel weapon against inflammation. Front. Pharmacol. 2021;12:622262. DOI: 10.3389/fphar.2021.622262.

7. Kristófi R., Eriksson J.W. Metformin as an anti-inflammatory agent: a short review. J. Endocrinol. 2021;251(2):R11–R22. DOI: 10.1530/JOE-21-0194.

8. Bu Y., Peng M., Tang X., Xu X., Wu Y., Chen A.F. et al. Protective effects of metformin in various cardiovascular diseases: Clinical evidence and AMPK-dependent mechanisms. J. Cell Mol. Med. 2022;26(19):4886–4903. DOI: 10.1111/jcmm.17519.

9. Aleidi S.M., Dahabiyeh L.A., Gu X., Al Dubayee M., Alshahrani A., Benabdelkamel H. et al. Obesity connected metabolic changes in type 2 diabetic patients treated with metformin. Front. Pharmacol. 2021;11:616157. DOI: 10.3389/fphar.2020.616157.

10. Dahabiyeh L.A., Mujammami M., Arafat T., Benabdelkamel H., Alfadda A.A., Abdel Rahman A.M. A metabolic pattern in healthy subjects given a single dose of metformin: a metabolomics approach. Front. Pharmacol. 2021;12:705932. DOI: 10.3389/fphar.2021.705932.

11. Dahabiyeh L.A., Mujammami M., AlMalki R.H., Arafat T., Benabdelkamel H., Alfadda A.A. et al. Lipids alterations associated with metformin in healthy subjects: an investigation using mass spectrometry shotgun approach. Int. J. Mol. Sci. 2022;23(19):11478. DOI: 10.3390/ijms231911478.

12. Mujammami M., Aleidi S.M., Buzatto A.Z., Alshahrani A., AlMalki R.H., Benabdelkamel H. et al. Lipidomics profiling of metformin-induced changes in obesity and type 2 diabetes mellitus: insights and biomarker potential. Pharmaceuticals (Basel). 2023;16(12):1717. DOI: 10.3390/ph16121717.

13. Zietzer A., Düsing P., Reese L., Nickenig G., Jansen F. Ceramide metabolism in cardiovascular disease: a network with high therapeutic potential. Arterioscler. Thromb. Vasc. Biol. 2022;42(10):1220–1228. DOI: 10.1161/ATVBAHA.122.318048.

14. Zhu Q., Scherer P.E. Ceramides and atherosclerotic cardiovascular disease: a current perspective. Circulation. 2024;149(21):1624–1626. DOI: 10.1161/CIRCULATIONAHA.123.065409.

15. Gruzdeva O.V., Akbasheva O.E., Dyleva Y.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 heart disease. Bull. Exp. Biol. Med. 2017;163(5):608–611. DOI: 10.1007/s10517-017-3860-5.

16. Turkistani A., Al-Kuraishy H.M., Al-Gareeb A.I., Alexiou A., Papadakis M., Bahaa M.M. et al. Pharmacological characterization of the antidiabetic drug metformin in atherosclerosis inhibition: A comprehensive insight. Immun. Inflamm. Dis. 2024;12(8):e1346. DOI: 10.1002/iid3.1346.

17. Delcheva G., Stefanova K., Stankova T. Ceramides-emerging biomarkers of lipotoxicity in obesity, diabetes, cardiovascular diseases, and inflammation. Diseases. 2024;12(9):195. DOI: 10.3390/diseases12090195.

18. Zabielski P., Chacinska M., Charkiewicz K., Baranowski M., Gorski J., Blachnio-Zabielska A.U. Effect of metformin on bioactive lipid metabolism in insulin-resistant muscle. J. Endocrinol. 2017;233(3):329–340. DOI: 10.1530/JOE-16-0381.

19. Grycel S., Markowski A.R., Hady H.R., Zabielski P., Kojta I., Imierska M. et al. Metformin treatment affects adipocytokine secretion and lipid composition in adipose tissues of diet-induced insulin-resistant rats. Nutrition. 2019;63-64:126–133. DOI: 10.1016/j.nut.2019.01.019.

20. Miklankova D., Markova I., Hüttl M., Zapletalova I., Poruba M., Malinska H. Metformin affects cardiac arachidonic acid metabolism and cardiac lipid metabolite storage in a prediabetic rat model. Int. J. Mol. Sci. 2021;22(14):7680. DOI: 10.3390/ijms22147680.

21. Hilvo M., Vasile V.C., Donato L.J., Hurme R., Laaksonen R. Ceramides and ceramide scores: clinical applications for cardiometabolic risk stratification. Front. Endocrinol. (Lausanne). 2020;11:570628. DOI: 10.3389/fendo.2020.570628.

22. Fujita K., Iwama H., Oura K., Tadokoro T., Hirose K., Watanabe M. et al. Metformin-suppressed differentiation of human visceral preadipocytes: Involvement of microRNAs. Int. J. Mol. Med. 2016;38(4):1135–1140. DOI: 10.3892/ijmm.2016.2729.