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Metabolomics Based on MS in Mice with Diet-Induced Obesity and Type 2 Diabetes Mellitus: the Effect of Vildagliptin, Metformin, and Their Combination

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Abstract

Type 2 diabetes mellitus (T2DM) is a major epidemiological problem. Metformin and vildagliptin are well-established antidiabetic drugs. The aim of the study was to evaluate the changes of plasma metabolic profile induced by a high-fat diet (HFD) and subsequent oral administration of metformin, vildagliptin, and their combination in a mouse model of diet-induced obesity (DIO)/T2DM analyzed using quadrupole-time-of-flight mass spectrometry (qTOF-MS). Metformin treatment increased the levels of butyrylcarnitine and acylcarnitine C18:1 concentrations and decreased the levels of isoleucine concentrations compared to untreated HFD mice. Vildagliptin treatment increased levels of butyrylcarnitine and acetylcarnitine. In summary, our metabolomics study revealed multiple differences between obese diabetic HFD mice and lean standard chow diet (SCD) mice, which were partially modifiable by subsequent metformin and vildagliptin treatment.

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References

  1. Lu, J., Xie, G., Jia, W., & Jia, W. (2013). Metabolomics in human type 2 diabetes research. Frontiers in Medicine, 7(1), 4–13. https://doi.org/10.1007/s11684-013-0248-4.

    Article  Google Scholar 

  2. Zhang, X., Imperatore, G., Thomas, W., Cheng, Y. J., Lobelo, F., Norris, K., Devlin, H. M., Ali, M. K., Gruss, S., Bardenheier, B., Cho, P., Garcia, I., Quevedo, D., Mudaliar, U., Saaddine, J., Geiss, L. S., & Gregg, E. W. (2016). Effect of lifestyle interventions on glucose regulation among adults without impaired glucose tolerance or diabetes: a systematic review and meta-analysis. Diabetes Res. Clin. Pract., 123, 149–164. https://doi.org/10.1016/j.diabres.2016.11.020.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wiernsperger, N. F., & Bailey, C. J. (1999). The antihyperglycaemic effect of metformin: therapeutic and cellular mechanisms. Drugs, 58(Suppl 1), 31–82.

    Article  CAS  PubMed  Google Scholar 

  4. Bailey, C. J., & Turner, R. C. (1996). Metformin. The New England Journal of Medicine, 334(9), 574–579. https://doi.org/10.1056/NEJM199602293340906.

    Article  CAS  PubMed  Google Scholar 

  5. Rena, G., Hardie, D. G., & Pearson, E. R. (2017). The mechanisms of action of metformin. Diabetologia, 60(9), 1577–1585. https://doi.org/10.1007/s00125-017-4342-z.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Inzucchi, S. E., Bergenstal, R. M., Buse, J. B., Diamant, M., Ferrannini, E., Nauck, M., Peters, A. L., Tsapas, A., Wender, R., & Matthews, D. R. (2015). Management of hyperglycemia in type 2 diabetes, 2015: a patient-centered approach. Update to a position statement of the American Diabetes Association and the European Association for the Study of diabetes. Diabetes Care, 38(1), 140–149.

    Article  PubMed  Google Scholar 

  7. Kim, H. J., Kim, J. H., Noh, S., Hur, H. J., Sung, M. J., Hwang, J. T., Park, J. H., Yang, H. J., Kim, M. S., Kwon, D. Y., & Yoon, S. H. (2010). Metabolomic analysis of livers and serum from high fat diet induced obese mice. Journal of Proteome Research, 10(2), 722–731. https://doi.org/10.1021/pr100892r.

    Article  CAS  PubMed  Google Scholar 

  8. Irving, B. A., Carter, R. E., Soop, M., Weymiller, A., Syed, H., Karakelides, H., Bhagra, S., Short, K. R., Tatpati, L., Barazzoni, R., & Nair, K. S. (2016). Effect of insulin sensitizer therapy on amino acids and their metabolites. Metabolism, 64, 720–728. https://doi.org/10.1016/j.metabol.2015.01.008.Effect.

    Article  Google Scholar 

  9. Huo, T., Cai, S., Lu, X., Sha, Y., Yu, M., & Li, F. (2009). Metabonomic study of biochemical changes in the serum of type 2 diabetes mellitus patients after the treatment of metformin hydrochloride. Journal of Pharmaceutical and Biomedical Analysis, 49(4), 976–982. https://doi.org/10.1016/j.jpba.2009.01.008.

    Article  CAS  PubMed  Google Scholar 

  10. Zhu, Y., Feng, Y., Shen, L., Xu, D., Wang, B., Ruan, K., & Cong, W. (2013). Effect of metformin on the urinary metabolites of diet-induced-obese mice studied by ultra performance liquid chromatography coupled to time-of-flight mass spectrometry (UPLC-TOF/MS). Journal of Chromatography B, 925, 110–116. https://doi.org/10.1016/j.jchromb.2013.02.040.

    Article  CAS  Google Scholar 

  11. Huo, T., Xiong, Z., Lu, X., & Cai, S. (2015). Metabonomic study of biochemical changes in urinary of type 2 diabetes mellitus patients after the treatment of sulfonylurea antidiabetic drugs based on ultra-performance liquid chromatography/mass spectrometry. Biomedical Chromatography, 29(1), 115–122. https://doi.org/10.1002/bmc.3247.

    Article  CAS  PubMed  Google Scholar 

  12. Qiu, Y., Rajagopalan, D., Connor, S. C., Damian, D., Zhu, L., Handzel, A., Hu, G., Amanullah, A., Bao, S., Woody, N., MacLean, D., Lee, K., Vanderwall, D., & Ryan, T. (2008). Multivariate classification analysis of metabolomic data for candidate biomarker discovery in type 2 diabetes mellitus. Metabolomics, 4(4), 337–346. https://doi.org/10.1007/s11306-008-0123-5.

    Article  CAS  Google Scholar 

  13. Pelantová, H., Bugáňová, M., Holubová, M., Šedivá, B., Zemenová, J., Sýkora, D., Kaválková, P., Haluzík, M., Železná, B., Maletínská, L., Kuneš, J., & Kuzma, M. (2016). Urinary metabolomic profiling in mice with diet-induced obesity and type 2 diabetes mellitus after treatment with metformin, vildagliptin and their combination. Mol. Cell. Endocrinol. https://doi.org/10.1016/j.mce.2016.05.003.

  14. Xu, J., Liu, C., & Cai, S. (2013). Metabolomic profilings of urine and serum from high fat-fed rats via 1 H NMR spectroscopy and pattern recognition. Applied Biochemistry and Biotechnology, 169(4), 1250–1261. https://doi.org/10.1007/s12010-012-0072-3.

    Article  CAS  PubMed  Google Scholar 

  15. Maletinska, L., Nagelova, V., Ticha, A., Zemenova, J., Pirnik, Z., Holubova, M., Spolcova, A., Mikulaskova, B., Blechova, M., Sykora, D., Lacinova, Z., Haluzik, M., Zelezna, B., & Kunes, J. (2015). Novel lipidized analogs of prolactin-releasing peptide have prolonged half-lives and exert anti-obesity effects after peripheral administration. International Journal of Obesity, 39(6), 986–993. https://doi.org/10.1038/ijo.2015.28.

    Article  CAS  PubMed  Google Scholar 

  16. Wu, T., Xie, G., Ni, Y., Liu, T., Yang, M., Wei, H., Jia, W., & Ji, G. (2015). Serum metabolite signatures of type 2 diabetes mellitus complications. Journal of Proteome Research, 14(1), 447–456. https://doi.org/10.1021/pr500825y.

    Article  CAS  PubMed  Google Scholar 

  17. Villarreal-Pérez, J. Z., Villarreal-Martínez, J. Z., Lavalle-González, F. J., Torres-Sepúlveda, M. D. R., Ruiz-Herrera, C., Cerda-Flores, R. M., Castillo-García, E. R., Rodríguez-Sánchez, I. P., & Martínez de Villarreal, L. E. (2014). Plasma and urine metabolic profiles are reflective of altered beta-oxidation in non-diabetic obese subjects and patients with type 2 diabetes mellitus. Diabetology and Metabolic Syndrome, 6(1), 129. https://doi.org/10.1186/1758-5996-6-129.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Balas, B., Baig, M. R., Watson, C., Dunning, B. E., Ligueros-Saylan, M., Wang, Y., He, Y. L., Darland, C., Holst, J. J., Deacon, C. F., Cusi, K., Mari, A., Foley, J. E., & DeFronzo, R. A. (2007). The dipeptidyl peptidase IV inhibitor vildagliptin suppresses endogenous glucose production and enhances islet function after single-dose administration in type 2 diabetic patients. The Journal of Clinical Endocrinology and Metabolism, 92(4), 1249–1255. https://doi.org/10.1210/jc.2006-1882.

    Article  CAS  PubMed  Google Scholar 

  19. Mari, A., Sallas, W. M., He, Y. L., Watson, C., Ligueros-Saylan, M., Dunning, B. E., Deacon, C. F., Holst, J. J., & Foley, J. E. (2005). Vildagliptin, a dipeptidyl peptidase-IV inhibitor, improves model-assessed ??-cell function in patients with type 2 diabetes. The Journal of Clinical Endocrinology and Metabolism, 90(8), 4888–4894. https://doi.org/10.1210/jc.2004-2460.

    Article  CAS  PubMed  Google Scholar 

  20. Lotfi, P., Yaghmaei, P., & Ebrahim-Habibi, A. (2015). Cymene and metformin treatment effect on biochemical parameters of male NMRI mice fed with high fat diet. Journal of Diabetes and Metabolic Disorders, 14(1), 52. https://doi.org/10.1186/s40200-015-0182-x.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Hong, A. R., Lee, J., Ku, E. J., Hwangbo, Y., Kim, K. M., Moon, J. H., Choi, S. H., Jang, H. C., & Lim, S. (2015). Comparison of vildagliptin as an add-on therapy and sulfonylurea dose-increasing therapy in patients with inadequately controlled type 2 diabetes using metformin and sulfonylurea (VISUAL study): a randomized trial. Diabetes Research and Clinical Practice, 109(1), 141–148. https://doi.org/10.1016/j.diabres.2015.04.019.

    Article  CAS  PubMed  Google Scholar 

  22. Halimi, S., Schweizer, A., Minic, B., Foley, J., & Dejager, S. (2008). Combination treatment in the management of type 2 diabetes: focus on vildagliptin and metformin as a single tablet. Vascular Health and Risk Management, 4(3), 481–492.

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Giesbertz, P., Padberg, I., Rein, D., Ecker, J., Höfle, A. S., Spanier, B., & Daniel, H. (2015). Metabolite profiling in plasma and tissues of ob/ob and db/db mice identifies novel markers of obesity and type 2 diabetes. Diabetologia, 58(9), 2133–2143. https://doi.org/10.1007/s00125-015-3656-y.

    Article  CAS  PubMed  Google Scholar 

  24. She, P., Olson, K. C., Kadota, Y., Inukai, A., Shimomura, Y., Hoppel, C. L., Adams, S. H., Kawamata, Y., Matsumoto, H., Sakai, R., Lang, C. H., & Lynch, C. J. (2013). Leucine and protein metabolism in obese Zucker rats. PLoS One, 8(3), e59443. https://doi.org/10.1371/journal.pone.0059443.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Lian, K., Du, C., Liu, Y., Zhu, D., Yan, W., Zhang, H., Hong, Z., Liu, P., Zhang, L., Pei, H., Zhang, J., Gao, C., Xin, C., Cheng, H., Xiong, L., & Tao, L. (2015). Impaired adiponectin signaling contributes to disturbed catabolism of branched-chain amino acids in diabetic mice. Diabetes, 64(1), 49–59. https://doi.org/10.2337/db14-0312.

    Article  CAS  PubMed  Google Scholar 

  26. Hawley, S. A., Gadalla, A. E., Olsen, G. S., & Hardie, D. G. (2002). The antidiabetic drug metformin activates the AMP-activated protein kinase cascade via an adenine nucleotide-independent mechanism. Diabetes, 51(8), 2420–2425. https://doi.org/10.2337/diabetes.51.8.2420.

    Article  CAS  PubMed  Google Scholar 

  27. Luo, P., Yu, H., Zhao, X., Bao, Y., Hong, C. S., Zhang, P., Tu, Y., Yin, P., Gao, P., Wei, L., Zhuang, Z., Jia, W., & Xu, G. (2016). Metabolomics study of Roux-en-Y gastric bypass surgery (RYGB) to treat type 2 diabetes patients based on ultraperformance liquid chromatography-mass spectrometry. Journal of Proteome Research, 15(4), 1288–1299. https://doi.org/10.1021/acs.jproteome.6b00022.

    Article  CAS  PubMed  Google Scholar 

  28. Garthwaite, T. L., Kalkhoff, R. K., Guansing, A. R., Hagen, T. C., & Menahan, L. A. (1979). Plasma free tryptophan, brain serotonin, and an endocrine profile of the genetically obese hyperglycemic mouse at 4–5 months of age. Endocrinology, 105, 1178–1182.

    Article  CAS  PubMed  Google Scholar 

  29. She, P., Van Horn, C., Reid, T., Hutson, S. M., Cooney, R. N., & Lynch, C. J. (2007). Obesity-related elevations in plasma leucine are associated with alterations in enzymes involved in branched-chain amino acid metabolism. American Journal of Physiology. Endocrinology and Metabolism, 293(6), 1552–1563. https://doi.org/10.1152/ajpendo.00134.2007.

    Article  CAS  Google Scholar 

  30. Xu, F., Tavintharan, S., Sum, C. F., Woon, K., Lim, S. C., & Ong, C. N. (2013). Metabolic signature shift in type 2 diabetes mellitus revealed by mass spectrometry-based metabolomics. The Journal of Clinical Endocrinology and Metabolism, 98(6), 1060–1065. https://doi.org/10.1210/jc.2012-4132.

    Article  CAS  Google Scholar 

  31. Koves, T. R., Ussher, J. R., Noland, R. C., Slentz, D., Mosedale, M., Ilkayeva, O., Bain, J., Stevens, R., Dyck, J. R. B., Newgard, C. B., Lopaschuk, G. D., & Muoio, D. M. (2008). Mitochondrial overload and incomplete fatty acid oxidation contribute to skeletal muscle insulin resistance. Cell Metabolism, 7(1), 45–56. https://doi.org/10.1016/j.cmet.2007.10.013.

    Article  CAS  PubMed  Google Scholar 

  32. Viollet, B., Guigas, B., Garcia, N. S., Leclerc, J., Foretz, M., & Andreelli, F. (2012). Cellular and molecular mechanisms of metformin: an overview. Clinical Science, 122(6), 253–270. https://doi.org/10.1042/CS20110386.

    Article  CAS  PubMed  Google Scholar 

  33. Srivastava, R. A. K., Pinkosky, S. L., Filippov, S., Hanselman, J. C., Cramer, C. T., & Newton, R. S. (2012). AMP-activated protein kinase: an emerging drug target to regulate imbalances in lipid and carbohydrate metabolism to treat cardio-metabolic diseases. Journal of Lipid Research, 53(12), 2490–2514. https://doi.org/10.1194/jlr.R025882.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Koonen, D. P. Y., Jacobs, R. L., Febbraio, M., Young, M. E., Soltys, C. L. M., Ong, H., Vance, D. E., & Dyck, J. R. B. (2007). Increased hepatic CD36 expression contributes to dyslipidemia associated with diet-induced obesity. Diabetes, 56(12), 2863–2871. https://doi.org/10.2337/db07-0907.

    Article  CAS  PubMed  Google Scholar 

  35. Schreurs, M., Kuipers, F., & Van Der Leij, F. R. (2010). Regulatory enzymes of mitochondrial beta-oxidation as targets for treatment of the metabolic syndrome. Obesity Reviews, 11(5), 380–388. https://doi.org/10.1111/j.1467-789X.2009.00642.x.

    Article  CAS  PubMed  Google Scholar 

  36. Aiyar, N., Disa, J., Ao, Z., Ju, H., Nerurkar, S., Willette, R. N., Macphee, C. H., Johns, D. G., & Douglas, S. A. (2007). Lysophosphatidylcholine induces inflammatory activation of human coronary artery smooth muscle cells. Molecular and Cellular Biochemistry, 295(1-2), 113–120. https://doi.org/10.1007/s11010-006-9280-x.

    Article  CAS  PubMed  Google Scholar 

  37. Schmitz, G., & Ruebsaamen, K. (2010). Metabolism and atherogenic disease association of lysophosphatidylcholine. Atherosclerosis, 208(1), 10–18. https://doi.org/10.1016/j.atherosclerosis.2009.05.029.

    Article  CAS  PubMed  Google Scholar 

  38. Shirouchi, B., Nagao, K., Inoue, N., Ohkubo, T., Hibino, H., & Yanagita, T. (2007). Effect of dietary omega 3 phosphatidylcholine on obesity-related disorders in obese Otsuka Long-Evans Tokushima fatty rats. Journal of Agricultural and Food Chemistry, 55(17), 7170–7176. https://doi.org/10.1021/jf071225x.

    Article  CAS  PubMed  Google Scholar 

  39. Sirdah, M. M., Abushahla, A. K., & Al-Sarraj, H. A. A. (2013). Effect of the addition of the antioxidant taurine on the complete blood count of whole blood stored at room temperature and at 4°C for up to 7 days. Revista Brasileira de Hematologia e Hemoterapia, 35(1), 44–51. https://doi.org/10.5581/1516-8484.20130014.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Wu, F., Koenig, K. L., Zeleniuch-Jacquotte, A., Jonas, S., Afanasyeva, Y., Wójcik, O. P., Costa, M., & Chen, Y. (2016). Serum taurine and stroke risk in women: a prospective, nested case-control study. PLoS One, 11(2), e0149348. https://doi.org/10.1371/journal.pone.0149348.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Ito, T., Yoshikawa, N., Ito, H., & Schaffer, S. W. (2015). Impact of taurine depletion on glucose control and insulin secretion in mice. Journal of Pharmacological Sciences, 129(1), 59–64. https://doi.org/10.1016/j.jphs.2015.08.007.

    Article  CAS  PubMed  Google Scholar 

  42. Pop-Busui, R., Sullivan, K. a., Van Huysen, C., Bayer, L., Cao, X., Towns, R., & Stevens, M. J. (2001). Depletion of taurine in experimental diabetic neuropathy: implications for nerve metabolic, vascular, and functional deficits. Experimental Neurology, 168(2), 259–272. https://doi.org/10.1006/exnr.2000.7591.

    Article  CAS  PubMed  Google Scholar 

  43. Ha, H., Yu, M. R., & Kim, K. H. (1999). Melatonin and taurine reduce early glomerulopathy in diabetic rats. Free Radical Biology & Medicine, 26, 944–950.

    Article  CAS  Google Scholar 

  44. Hansen, S. H. (2001). The role of taurine in diabetes and the development of diabetic complications. Diabetes/Metabolism Research and Reviews, 17(5), 330–346. https://doi.org/10.1002/dmrr.229.

    Article  CAS  PubMed  Google Scholar 

  45. Yu, H., Guo, Z., Shen, S., & Shan, W. (2016). Effects of taurine on gut microbiota and metabolism in mice. Amino Acids, 48(7), 1601–1617. https://doi.org/10.1007/s00726-016-2219-y.

    Article  CAS  PubMed  Google Scholar 

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Funding

This research was financially supported by the Grant Agency of the Czech Republic [Grant No. GA13-14105S] and Programme to support medical applied research, Ministry of Health, Czech Republic 2015-2022 [NV15-26854A]. This project was conducted within the “Prague Infrastructure for Structure Biology and Metabolomics” with the financial support of the Operational Program Prague – Competitiveness [Project No. CZ.2.16/3.1.00/24023] and institutional support [RVO 61388963, 67985823, 61388971, and RVO VFN64165]. The authors would also like to acknowledge project LO1509 of the Ministry of Education, Youth and Sports of the Czech Republic for financial support.

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Authors and Affiliations

  1. Institute of Microbiology, The Czech Academy of Sciences, Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Petra Tomášová, Martina Bugáňová, Helena Pelantová & Marek Kuzma

  2. Fourth Medical Department, First Faculty of Medicine, Charles University in Prague and General University Hospital, U nemocnice 1, 128 08, Prague 2, Czech Republic

    Petra Tomášová

  3. Faculty of Chemical Technology, University of Chemistry and Technology Prague, Technická 5, 166 28, Prague 6, Czech Republic

    Martina Bugáňová

  4. Institute of Organic Chemistry and Biochemistry, The Czech Academy of Sciences, Flemingovo nám. 2, 166 10, Prague 6, Czech Republic

    Martina Holubová, Blanka Železná, Lenka Maletínská & Jaroslav Kuneš

  5. Faculty of Applied Sciences, University of West Bohemia, Univerzitní 8, 306 14, Pilsen, Czech Republic

    Blanka Šedivá

  6. Institute of Medical Biochemistry and Laboratory Diagnostics; First Faculty of Medicine, Charles University in Prague and General University Hospital, U nemocnice 2, 128 08, Prague 2, Czech Republic

    Martin Haluzík

  7. Centre for Experimental Medicine, Institute for Clinical and Experimental Medicine, Vídeňská 1958/9, 140 21, Prague 4, Czech Republic

    Martin Haluzík

  8. Institute of Physiology, The Czech Academy of Sciences, Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Jaroslav Kuneš

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  1. Petra Tomášová
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  2. Martina Bugáňová
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Correspondence to Marek Kuzma.

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All experiments were conducted according to the ethical guidelines for animal experiments and the Czech Republic law no. 246/1992 and were approved through the Committee for Experiments with Laboratory Animals of the Academy of Sciences of the Czech Republic.

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The authors declare that they have no conflict of interest.

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Highlights

• The differences in plasma metabolomics profile in DIO mouse were described

• The changes in plasma metabolomics profile after antidiabetic treatment were studied

• The combination of antidiabetic drugs restored levels of leucine and tryptophan

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Tomášová, P., Bugáňová, M., Pelantová, H. et al. Metabolomics Based on MS in Mice with Diet-Induced Obesity and Type 2 Diabetes Mellitus: the Effect of Vildagliptin, Metformin, and Their Combination. Appl Biochem Biotechnol 188, 165–184 (2019). https://doi.org/10.1007/s12010-018-2899-8

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