Short and long term in vivo effects of Cyclosporine A and Sirolimus on genes and proteins involved in lipid metabolism in Wistar rats Patrícia C. Lopes a , Amelia Fuhrmann a , José Sereno b, c , Daniel O. Espinoza a , Maria João Pereira a, d , Jan W. Eriksson d , Flávio Reis b , Eugenia Carvalho a, e , ⁎ a Center for Neuroscience and Cell Biology, University of Coimbra, 3000-517 Coimbra, Portugal b Laboratory of Pharmacology & Experimental Therapeutics, IBILI, Faculty of Medicine, University of Coimbra, 3000-548 Coimbra, Portugal c Institute for Nuclear Sciences Applied to Heath–ICNAS, University of Coimbra, 3000-548 Coimbra, Portugal d Department of Medical Sciences, Uppsala University, 751 85 Uppsala, Sweden e The Portuguese Diabetes Association (APDP-ERC), 1250 203 Lisbon, Portugal ARTICLE INFO ABSTRACT Article history: Received 6 November 2013 Accepted 6 February 2014 Objective. Cyclosporine A (CsA) and sirolimus (SRL) are immunosuppressive agents (IA) associated with new onset diabetes after transplantation and dyslipidemia. We aim to evaluate the molecular effects of CsA (5 mg/kg/day) and SRL (1 mg/kg/day) treatment for 3 and 9 weeks on lipid metabolism, in Wistar rats. Materials/Methods. Lipolysis was evaluated in isolated adipocytes, while triglycerides (TG) and non-esterified fatty acid (NEFA) were measured in serum. Gene and protein expression involved in lipid metabolism was assessed in adipose tissue and liver. Results. CsA and SRL treatments of rats for 3 and 9 weeks increased isoproterenol- stimulated lipolysis by 5–9 fold and 4–6 fold in isolated adipocytes, respectively. While CsA increased adipocyte weight and diameter, as well as NEFA and TG levels in circulation after 9 weeks, SRL treatment caused ectopic deposition of TG in the liver after 3 weeks. Moreover, ACC1 and FAS protein expression was increased after 3 weeks (>100%, p < 0.01), while HSL was increased after 9 weeks of CsA treatment. On the other hand, SRL decreased the expression of lipogenic genes, including ACC1 (50%, p < 0.05), lipin1 (25%, p < 0.05), PPAR-γ (42%, p < 0.05) and SCD1 (80%, p < 0.001) in adipose tissue, after 3 weeks of treatment. Conclusion. The effects of both IAs on expression of lipolytic and lipogenic genes suggest that these agents influence lipid metabolism, thus contributing to the dyslipidemia observed during immunosuppressive therapy. © 2014 Elsevier Inc. All rights reserved. Keywords: Immunosuppressive therapy Adipocytes Liver Dyslipidemia METABOLISM CLINICAL AND EXPERIMENTAL 63 (2014) 702 – 715 Abbreviations: ACC, Acetyl-CoA carboxylase; ATGL, Adipose triacylglycerol lipase; CD36, Fatty acid translocase; CsA, Cyclosporine A; CHREBP, Carbohydrate response element-binding protein; DGAT1, Diglyceride acyltransferase 1; FAS, Fatty acid synthase; FFA, Free Fatty Acid; HSL, Hormone-sensitive lipase; IA, Immunosuppressive agent; IL-6, Interleukin-6; LDL, Low-density lipoprotein; LPL, Lipoprotein lipase; mTOR, Mammalian target of rapamycin; NEFA, non-esterified fatty acid; NODAT, New onset diabetes after transplantation; PPARγ, Peroxisome proliferator-activated receptor γ; SCD1, Stearoyl-CoA desaturase 1; SRL, Sirolimus; SREBP, Sterol Regulatory Element-Binding Protein; TBP, TATA binding protein; TG, Triglycerides; TNF-α, Tumor necrosis factor -α; VLDL, Very low density lipoprotein. ⁎ Corresponding author at: Center for Neurosciences and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal. Tel.: + 351 239 853 406; fax: +351 239 853 1409. E-mail address: ecarvalh@cnc.uc.pt (E. Carvalho). 0026-0495/$ – see front matter © 2014 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.metabol.2014.02.004 Available online at www.sciencedirect.com Metabolism www.metabolismjournal.com