Skip to main content Accessibility help

Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age

  • Uzman B. Cheema (a1), Erika Most (a1), Klaus Eder (a1) and Robert Ringseis (a1)


While strong evidence from clinical studies suggests beneficial effects of carnitine supplementation on metabolic health, serious safety concerns associated with carnitine supplementation have been raised from studies in mice. Considering that the carnitine doses in these mice studies were up to 100 times higher than those used in clinical studies, the present study aimed to address possible safety concerns associated with long-term supplementation of a carnitine dose used in clinical trials. Two groups of NMRI mice were fed either a control or a carnitine-supplemented diet (1 g/kg diet) from weaning to 19 months of age, and parameters of hepatic lipid metabolism and stress signalling and skeletal muscle gene expression were analysed in the mice at 19 months of age. Concentrations of free carnitine and acetylcarnitine in plasma and tissues were higher in the carnitine than in the control group (P<0·05). Plasma concentrations of free carnitine and acetylcarnitine were higher in mice at adult age (10 and 15 months) than at advanced age (19 months) (P<0·05). Hepatic mRNA and protein levels of genes involved in lipid metabolism and stress signalling and hepatic and plasma lipid concentrations did not differ between the carnitine and the control group. Skeletal muscle transcriptome analysis in 19-month-old mice revealed only a moderate regulation between carnitine and control group. Lifelong carnitine supplementation prevents an age-dependent impairment of plasma carnitine status, but safety concerns associated with long-term supplementation of carnitine at doses used in clinical trials can be considered as unfounded.

  • View HTML
    • Send article to Kindle

      To send this article to your Kindle, first ensure is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the or variations. ‘’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age
      Available formats

      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age
      Available formats

      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age
      Available formats


This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (, which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

Corresponding author

*Corresponding author: R. Ringseis, email


Hide All
1. Fritz, IB & McEwen, B (1959) Effects of carnitine on fatty-acid oxidation by muscle. Science 129, 334335.
2. Lysiak, W, Lilly, K, DiLisa, F, et al. (1988) Quantitation of the effect of l-carnitine on the levels of acid-soluble short-chain acyl-CoA and CoASH in rat heart and liver mitochondria. J Biol Chem 263, 11511156.
3. Uziel, G, Garavaglia, B & Di Donato, S (1988) Carnitine stimulation of pyruvate dehydrogenase complex (PDHC) in isolated human skeletal muscle mitochondria. Muscle Nerve 11, 720724.
4. Broderick, TL, Quinney, HA & Lopaschuk, GD (1992) Carnitine stimulation of glucose oxidation in the fatty acid perfused isolated working rat heart. J Biol Chem 267, 37583763.
5. De Palo, E, Gatti, R, Sicolo, N, et al. (1981) Plasma and urine free l-carnitine in human diabetes mellitus. Acta Diabetol Lat 18, 9195.
6. Pregant, P, Schernthaner, G, Legenstein, E, et al. (1991) Decreased plasma carnitine in type I diabetes mellitus. Klin Wochenschr 69, 511516.
7. Poorabbas, A, Fallah, F, Bagdadchi, J, et al. (2007) Determination of free l-carnitine levels in type II diabetic women with and without complications. Eur J Clin Nutr 61, 892895.
8. Ringseis, R, Keller, J & Eder, K (2012) Role of carnitine in the regulation of glucose homeostasis and insulin sensitivity: evidence from in vivo and in vitro studies with carnitine supplementation and carnitine deficiency. Eur J Nutr 51, 118.
9. Waber, LJ, Valle, D, Neill, C, et al. (1982) Carnitine deficiency presenting as familial cardiomyopathy: a treatable defect in carnitine transport. J Pediatr 101, 700705.
10. Glasgow, AM, Engel, AG, Bier, DM, et al. (1983) Hypoglycemia, hepatic dysfunction, muscle weakness, cardiomyopathy, free carnitine deficiency and long-chain acylcarnitine excess responsive to medium chain triglyceride diet. Pediatr Res 17, 319326.
11. Tripp, ME, Katcher, ML, Peters, HA, et al. (1981) Systemic carnitine deficiency presenting as familial endocardial fibroelastosis: a treatable cardiomyopathy. N Engl J Med 305, 385390.
12. Gramignano, G, Lusso, MR, Madeddu, C, et al. (2006) Efficacy of l-carnitine administration on fatigue, nutritional status, oxidative stress, and related quality of life in 12 advanced cancer patients undergoing anticancer therapy. Nutrition 22, 136145.
13. Steiber, AL, Davis, AT, Spry, L, et al. (2006) Carnitine treatment improved quality-of-life measure in a sample of Midwestern hemodialysis patients. JPEN J Parenter Enteral Nutr 30, 1015.
14. Mantovani, G, Macciò, A, Madeddu, C, et al. (2008) Randomized phase III clinical trial of five different arms of treatment for patients with cancer cachexia: interim results. Nutrition 24, 305313.
15. Malaguarnera, M, Vacante, M, Giordano, M, et al. (2011) Oral acetyl-l-carnitine therapy reduces fatigue in overt hepatic encephalopathy: a randomized, double-blind, placebo-controlled study. Am J Clin Nutr 93, 799808.
16. Rabito, EI, Leme, IA, Demenice, R, et al. (2013) Lower carnitine plasma values from malnutrition cancer patients. J Gastrointest Cancer 44, 362365.
17. De Simone, C, Tzantzoglou, S, Jirillo, E, et al. (1992) l-Carnitine deficiency in AIDS patients. AIDS 6, 203205.
18. Vinci, E, Rampello, E, Zanoli, L, et al. (2005) Serum carnitine levels in patients with tumoral cachexia. Eur J Intern Med 16, 419423.
19. Hockenberry, MJ, Hooke, MC, Gregurich, M, et al. (2009) Carnitine plasma levels and fatigue in children/adolescents receiving cisplatin, ifosfamide, or doxorubicin. J Pediatr Hematol Oncol 31, 664669.
20. Koeth, RA, Wang, Z, Levison, BS, et al. (2013) Intestinal microbiota metabolism of l-carnitine, a nutrient in red meat, promotes atherosclerosis. Nat Med 19, 576585.
21. Lu, Y, Li, W & Yang, X (2017) Soluble soybean polysaccharides enhance the protective effects of genistein against hepatic injury in high l-carnitine-fed mice. Food Funct 8, 43644373.
22. Brass, EP (2000) Supplemental carnitine and exercise. Am J Clin Nutr 72 Suppl, 618S623S.
23. Ringseis, R, Keller, J & Eder, K (2013) Mechanisms underlying the anti-wasting effect of l-carnitine supplementation under pathologic conditions: evidence from experimental and clinical studies. Eur J Nutr 52, 14211442.
24. NRC (National Research Council) (2011) Guide for the Care and Use of Laboratory Animals, 8th ed. Washington, DC: National Academies Press.
25. NRC (National Research Council) (1995) Nutrient Requirements of Laboratory Animals, 4th revised ed. Washington, DC: National Academies Press.
26. Hirche, F, Fischer, M, Keller, J, et al. (2009) Determination of carnitine, its short chain acyl esters and metabolic precursors trimethyllysine and gamma-butyrobetaine by quasi-solid phase extraction and MS/MS detection. J Chromatogr B Analyt Technol Biomed Life Sci 877, 21582162.
27. Hara, A & Radin, NS (1978) Lipid extraction of tissues with a low-toxicity solvent. Anal Biochem 90, 420426.
28. Eder, K & Kirchgessner, M (1994) Dietary fat influences the effect of zinc deficiency on liver lipids and fatty acids in rats force-fed equal quantities of diet. J Nutr 124, 19171926.
29. Chiappisi, E, Ringseis, R, Eder, K, et al. (2017) Effect of endoplasmic reticulum stress on metabolic and stress signaling and kidney-specific functions in Madin–Darby bovine kidney cells. J Dairy Sci 100, 66896706.
30. Vandesompele, J, De Preter, K, Pattyn, F, et al. (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3, RESEARCH0034.
31. Ringseis, R, Mooren, FC, Keller, J, et al. (2011) Regular endurance exercise improves the diminished hepatic carnitine status in mice fed a high-fat diet. Mol Nutr Food Res 55 Suppl. 2, S193S202.
32. Gessner, DK, Winkler, A, Koch, C, et al. (2017) Analysis of hepatic transcript profile and plasma lipid profile in early lactating dairy cows fed grape seed and grape marc meal extract. BMC Genomics 18, 253.
33. Edgar, R, Domrachev, M & Lash, AE (2002) Gene Expression Omnibus: NCBI gene expression and hybridization array data repository. Nucleic Acids Res 30, 207210.
34. Irizarry, RA, Hobbs, B, Collin, F, et al. (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249264.
35. Huang, DW, Sherman, BT & Lempicki, RA (2009) Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 37, 113.
36. Huang, DW, Sherman, BT & Lempicki, RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4, 4457.
37. Morand, R, Bouitbir, J, Felser, A, et al. (2014) Effect of carnitine, acetyl-, and propionylcarnitine supplementation on the body carnitine pool, skeletal muscle composition, and physical performance in mice. Eur J Nutr 53, 13131325.
38. Ringseis, R, Lüdi, S, Hirche, F, et al. (2008) Treatment with pharmacological peroxisome proliferator-activated receptor alpha agonist clofibrate increases intestinal carnitine absorption in rats. Pharmacol Res 58, 5864.
39. Lambert, BD, Dobson, CM, Cherry, NM, et al. (2009) Chemical form of dietary l-carnitine affects plasma but not tissue carnitine concentrations in male Sprague–Dawley rats. J Anim Physiol Anim Nutr (Berl) 93, 174180.
40. Barnett, C, Costill, DL, Vukovich, MD, et al. (1994) Effect of l-carnitine supplementation on muscle and blood carnitine content and lactate accumulation during high-intensity sprint cycling. Int J Sport Nutr 4, 280288.
41. Wächter, S, Vogt, M, Kreis, R, et al. (2002) Long-term administration of l-carnitine to humans: effect on skeletal muscle carnitine content and physical performance. Clin Chim Acta 318, 5161.
42. Rebouche, CJ (1977) Carnitine movement across muscle cell membranes. Studies in isolated rat muscle. Biochim Biophys Acta 471, 145155.
43. Tamai, I, Ohashi, R, Nezu, J, et al. (1998) Molecular and functional identification of sodium ion-dependent, high affinity human carnitine transporter OCTN(2). J Biol Chem 273, 2037820382.
44. Stephens, FB, Constantin-Teodosiu, D, Laithwaite, D, et al. (2006) Insulin stimulates l-carnitine accumulation in human skeletal muscle. FASEB J 20, 377379.
45. Wall, BT, Stephens, FB, Constantin-Teodosiu, D, et al. (2011) Chronic oral ingestion of l-carnitine and carbohydrate increases muscle carnitine content and alters muscle fuel metabolism during exercise in humans. J Physiol 589, 963973.
46. Stephens, FB, Constantin-Teodosiu, D & Greenhaff, PL (2007) New insights concerning the role of carnitine in the regulation of fuel metabolism in skeletal muscle. J Physiol 581, 431444.
47. Fischer, M, Varady, J, Hirche, F, et al. (2009) Supplementation of l-carnitine in pigs: absorption of carnitine and effect on plasma and tissue carnitine concentrations. Arch Anim Nutr 63, 115.
48. Kaup, D, Keller, J, Most, E, et al. (2018) The carnitine status does not affect the contractile and metabolic phenotype of skeletal muscle in pigs. Nutr Metab (Lond) 15, 2.
49. Kato, Y, Sugiura, M, Sugiura, T, et al. (2006) Organic cation/carnitine transporter OCTN2 (Slc22a5) is responsible for carnitine transport across apical membranes of small intestinal epithelial cells in mouse. Mol Pharmacol 70, 829837.
50. Tamai, I, China, K, Sai, Y, et al. (2001) Na(+)-coupled transport of l-carnitine via high-affinity carnitine transporter OCTN2 and its subcellular localization in kidney. Biochim Biophys Acta 1512, 273284.
51. García-Delgado, M, Peral, MJ, Durán, JM, et al. (2009) Ontogeny of Na(+)/l-carnitine transporter and of gamma-trimethylaminobutyraldehyde dehydrogenase and gamma-butyrobetaine hydroxylase genes expression in rat kidney. Mech Ageing Dev 130, 227233.
52. García-Miranda, P, Durán, JM, Peral, MJ, et al. (2005) Developmental maturation and segmental distribution of rat small intestinal l-carnitine uptake. J Membr Biol 206, 916.
53. Karlic, H, Lohninger, A, Laschan, C, et al. (2003) Downregulation of carnitine acyltransferases and organic cation transporter OCTN2 in mononuclear cells in healthy elderly and patients with myelodysplastic syndromes. J Mol Med (Berl) 81, 435442.
54. Vaz, FM & Wanders, RJ (2002) Carnitine biosynthesis in mammals. Biochem J 361, 417429.
55. Rebouche, CJ, Lehman, LJ & Olson, L (1986) Epsilon-N-trimethyllysine availability regulates the rate of carnitine biosynthesis in the growing rat. J Nutr 116, 751759.
56. Vernez, L, Dickenmann, M, Steiger, J, et al. (2006) Effect of l-carnitine on the kinetics of carnitine, acylcarnitines and butyrobetaine in long-term haemodialysis. Nephrol Dial Transplant 21, 450458.
57. Primassin, S & Spiekerkoetter, U (2010) ESI-MS/MS measurement of free carnitine and its precursor γ-butyrobetaine in plasma and dried blood spots from patients with organic acidurias and fatty acid oxidation disorders. Mol Genet Metab 101, 141145.
58. Rebouche, CJ (1983) Effect of dietary carnitine isomers and γ-butyrobetaine on l-carnitine biosynthesis and metabolism in the rat. J Nutr 113, 19061913.
59. Koeth, RA, Levison, BS, Culley, MK, et al. (2014) γ-Butyrobetaine is a proatherogenic intermediate in gut microbial metabolism of l-carnitine to TMAO. Cell Metab 20, 799812.
60. Rebouche, CJ, Mack, DL & Edmonson, PF (1984) l-Carnitine dissimilation in the gastrointestinal tract of the rat. Biochemistry 23, 64226426.
61. Seim, H, Schulze, J & Strack, E (1985) Catabolic pathways for high-dosed l(−)- or d(+)-carnitine in germ-free rats? Biol Chem Hoppe Seyler 366, 10171021.
62. Cullinan, SB, Zhang, D, Hannink, M, et al. (2003) Nrf2 is a direct PERK substrate and effector of PERK-dependent cell survival. Mol Cell Biol 23, 71987209.
63. Cnop, M, Foufelle, F & Velloso, LA (2012) Endoplasmic reticulum stress, obesity and diabetes. Trends Mol Med 18, 5968.
64. Pagliassotti, MJ, Kim, PY, Estrada, AL, et al. (2016) Endoplasmic reticulum stress in obesity and obesity-related disorders: An expanded view. Metabolism 65, 12381246.
65. Hoyte, K, Kang, C & Martin, PT (2002) Definition of pre- and postsynaptic forms of the CT carbohydrate antigen at the neuromuscular junction: ubiquitous expression of the CT antigens and the CT GalNAc transferase in mouse tissues. Brain Res Mol Brain Res 109, 146160.
66. Thomas, PJ, Xu, R & Martin, PT (2016) B4GALNT2 (GALGT2) gene therapy reduces skeletal muscle pathology in the FKRP P448L mouse model of limb girdle muscular dystrophy 2I. Am J Pathol 186, 24292448.
67. Kemp, TJ, Sadusky, TJ, Saltisi, F, et al. (2000) Identification of ankrd2, a novel skeletal muscle gene coding for a stretch-responsive ankyrin-repeat protein. Genomics 66, 229241.
68. Tsukamoto, Y, Senda, T, Nakano, T, et al. (2002) Arpp, a new homolog of carp, is preferentially expressed in type 1 skeletal muscle fibers and is markedly induced by denervation. Lab Invest 82, 645655.
69. Lehti, M, Kivela, R, Komi, P, et al. (2009) Effects of fatiguing jumping exercise on mRNA expression of titin-complex proteins and calpains. J Appl Physiol 106, 14191424.
70. Fleischer, J, Breer, H & Strotmann, J (2009) Mammalian olfactory receptors. Front Cell Neurosci 3, 9.
71. Kang, N & Koo, J (2012) Olfactory receptors in non-chemosensory tissues. BMB Rep 45, 612622.
72. Pichavant, C, Burkholder, TJ & Pavlath, GK (2016) Decrease of myofiber branching via muscle-specific expression of the olfactory receptor mOR23 in dystrophic muscle leads to protection against mechanical stress. Skelet Muscle 6, 2.
73. Kalbe, B, Osterloh, M, Schulz, VM, et al. (2018) OR2H2 regulates the differentiation of human myoblast cells by its ligand aldehyde 13-(13). Arch Biochem Biophys 645, 7280.
74. Wang, Z, Klipfell, E, Bennett, BJ, et al. (2011) Gut flora metabolism of phosphatidylcholine promotes cardiovascular disease. Nature 472, 5763.
75. Bennett, BJ, de Aguiar Vallim, TQ, Wang, Z, et al. (2013) Trimethylamine-N-oxide, a metabolite associated with atherosclerosis, exhibits complex genetic and dietary regulation. Cell Metab 17, 4960.


Type Description Title
Supplementary materials

Cheema et al. supplementary material
Tables S1-S8 and Figures S1-S3

 Word (319 KB)
319 KB

Effect of lifelong carnitine supplementation on plasma and tissue carnitine status, hepatic lipid metabolism and stress signalling pathways and skeletal muscle transcriptome in mice at advanced age

  • Uzman B. Cheema (a1), Erika Most (a1), Klaus Eder (a1) and Robert Ringseis (a1)


Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed