Abstract
Mildronate (MD) is a cardioprotective drug used for the treatment of cardiovascular diseases by switching metabolism from the fatty acids to glucose oxidation. This effect is achieved via inhibition of synthesis of L-carnitine (L-car), a common supplement, which is used for improving of fatty acid metabolism. Both MD and L-car have similar neuroprotective effect. Our goal was to investigate the effect of two drugs on the cognitive parameters of mice under different conditions (aging and lipopolysaccharide (LPS)-induced inflammation). We showed that L-car partly improved the memory and decreased the extent of mtDNA damage in the hippocampus of mice with the LPS-induced inflammation. L-car induced mitochondrial biogenesis and mitophagy in the Nrf2-dependent manner. Both MD and L-car upregulated expression of genes involved in the mitochondrial quality control. In 15-month-old mice, MD improved long-term and short-term memory, reduced the extent of mtDNA damage, and decreased the concentration of diene conjugates in the hippocampus in the Nrf2-independent manner. L-car as a Nrf2 activator had a better neuroprotective effect by normalizing mitochondrial quality control in the reversible cognitive impairment caused by the LPS-induced inflammation, while MD had a better neuroprotective effect in the irreversible cognitive impairment in aged mice, possibly due to a deeper restructuring of metabolism and reduction of oxidative stress.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data presented in this study are available on request.
References
Ackert-Bicknell CL, Anderson LC, Sheehan S, Hill WG, Chang B, Churchill GA, Chesler EJ, Korstanje R, Peters LL (2015) Aging research using mouse models. Curr Protoc Mouse Biol 5:95–133. https://doi.org/10.1002/9780470942390.mo140195
Alzoubi KH, Rababa’h AM, Owaisi A, Khabour OF (2017) L-carnitine prevents memory impairment induced by chronic REM-sleep deprivation. Brain Res Bull 131:176–182. https://doi.org/10.1016/j.brainresbull.2017.04.004
Anaeigoudari A, Shafei MN, Soukhtanloo M, Sadeghnia HR, Reisi P, Beheshti F, Mohebbati R, Mousavi SM, Hosseini M (2015) Lipopolysaccharide-induced memory impairment in rats is preventable using 7-nitroindazole. Arq Neuropsiquiatr 73:784–790. https://doi.org/10.1590/0004-282X20150121
Asaka N, Muranaka Y, Kirimoto T, Miyake H (1998) Cardioprotective profile of MET-88, an inhibitor of carnitine synthesis, and insulin during hypoxia in isolated perfused rat hearts. Fundam Clin Pharmacol 12:158–63. https://doi.org/10.1111/j.1472-8206.1998.tb00936.x
Beitnere U, van Groen T, Kumar A, Jansone B, Klusa V, Kadish I (2014) Mildronate improves cognition and reduces amyloid-β pathology in transgenic Alzheimer’s disease mice. J Neurosci Res 92:338–346. https://doi.org/10.1002/jnr.23315
Bilbo SD, Rudy JW, Watkins LR, Maier SF (2006) A behavioural characterization of neonatal infection-facilitated memory impairment in adult rats. Behav Brain Res 169:39–47. https://doi.org/10.1016/j.bbr.2005.12.002
Binienda Z, Przybyla-Zawislak B, Virmani A, Schmued L (2005) L-carnitine and neuroprotection in the animal model of mitochondrial dysfunction. Ann N Y Acad Sci 1053:174–182. https://doi.org/10.1196/annals.1344.015
Block BR, Albanese SG, Hume AL (2021) Online promotion of “Brain Health” supplements. Sr Care Pharm 36:489–492. https://doi.org/10.4140/TCP.n.2021.489
Chen N, Yang M, Zhou M, Xiao J, Guo J, He L (2017) L-carnitine for cognitive enhancement in people without cognitive impairment. Cochrane Database Syst Rev 3:CD009374. https://doi.org/10.1002/14651858.CD009374.pub3
Dambrova M, Liepinsh E, Kalvinsh I (2002) Mildronate: cardioprotective action through carnitine-lowering effect. Trends Cardiovasc Med 12:275–279. https://doi.org/10.1016/s1050-1738(02)00175-5
Di Cristo F, Calarco A, Digilio FA, Sinicropi MS, Rosano C, Galderisi U, Beatrice Melone MA, Saturnino C, Peluso G (2020) The discovery of highly potent THP derivatives as OCTN2 inhibitors: From structure-based virtual screening to in vivo biological activity. Int J Mol Sci 21:7431. https://doi.org/10.3390/ijms21197431
Di Cristo F, Finicelli M, Digilio FA, Paladino S, Valentino A, Scialò F, D’Apolito M, Saturnino C, Galderisi U, Giordano A, Melone MAB, Peluso G (2018) Meldonium improves Huntington’s disease mitochondrial dysfunction by restoring peroxisome proliferator-activated receptor γ coactivator 1α expression. J Cell Physiol 234:9233–9246. https://doi.org/10.1002/jcp.27602
Fang EF, Scheibye-Knudsen M, Chua KF, Mattson MP, Croteau DL, Bohr VA (2016) Nuclear DNA damage signalling to mitochondria in ageing. Nat Rev Mol Cell Biol 17:308–321. https://doi.org/10.1038/nrm.2016.14
Ferreira GC, McKenna MC (2017) L-Carnitine and acetyl-L-carnitine roles and neuroprotection in developing brain. Neurochem Res 42:1661–1675. https://doi.org/10.1007/s11064-017-2288-7
Gorelick PB, Counts SE, Nyenhuis D (2016) Vascular cognitive impairment and dementia. Biochim Biophys Acta 1862:860–868. https://doi.org/10.1016/j.bbadis.2015.12.015
Gülçin İ (2006) Antioxidant and antiradical activities of L-carnitine. Life Sci 78:803–11. https://doi.org/10.1016/j.lfs.2005.05.103
Gureev AP, Shaforostova EA, Starkov AA, Popov VN (2017) Simplified qPCR method for detecting excessive mtDNA damage induced by exogenous factors. Toxicology 382:67–74. https://doi.org/10.1016/j.tox.2017.03.010
Ji MH, Zhang L, Mao MJ, Zhang H, Yang JJ, Qiu LL (2020) Overinhibition mediated by parvalbumin interneurons might contribute to depression-like behavior and working memory impairment induced by lipopolysaccharide challenge. Behav Brain Res 383:112509. https://doi.org/10.1016/j.bbr.2020.112509
Kepka A, Ochocinska A, Borzym-Kluczyk M, Skorupa E, Stasiewicz-Jarocka B, Chojnowska S, Waszkiewicz N (2020) Preventive role of L-carnitine and balanced diet in Alzheimer’s disease. Nutrients 12:1987. https://doi.org/10.3390/nu12071987
Kerr JS, Adriaanse BA, Greig NH, Mattson MP, Cader MZ, Bohr VA, Fang EF (2017) Mitophagy and Alzheimer’s disease: cellular and molecular mechanisms. Trends Neurosci 40:151–166. https://doi.org/10.1016/j.tins.2017.01.002
Khan MS, Muhammad T, Ikram M, Kim MO (2019) Dietary supplementation of the antioxidant curcumin halts systemic LPS-induced neuroinflammation-associated neurodegeneration and memory/synaptic impairment via the JNK/NF-κB/Akt signaling pathway in adult rats. Oxid Med Cell Longev 2019:7860650. https://doi.org/10.1155/2019/7860650
Klusa V, Beitnere U, Pupure J, Isajevs S, Rumaks J, Svirskis S, Dzirkale Z, Kalvinsh I (2013) Mildronate and its neuroregulatory mechanisms: targeting the mitochondria, neuroinflammation, and protein expression. Med (Kaunas) 49:301–309
Klusa VZ, Isajevs S, Svirina D, Pupure J, Beitnere U, Rumaks J, Svirskis S, Jansone B, Dzirkale Z, Muceniece R, Kalvinsh I, Vinters HV (2010) Neuroprotective properties of mildronate, a small molecule, in a rat model of Parkinson’s disease. Int J Mol Sci 11:4465–4487. https://doi.org/10.3390/ijms11114465
Koch M, Fitzpatrick AL, Rapp SR, Nahin RL, Williamson JD, Lopez OL, DeKosky ST, Kuller LH, Mackey RH, Mukamal KJ, Jensen MK, Sink KM (2019) Alcohol consumption and risk of dementia and cognitive decline among older adults with or without mild cognitive impairment. JAMA Netw open 2:e1910319–e1910319. https://doi.org/10.1001/jamanetworkopen.2019.10319
Kuka J, Vilskersts R, Cirule H, Makrecka M, Pugovics O, Kalvinsh I, Dambrova M, Liepinsh E (2012) The cardioprotective effect of mildronate is diminished after co-treatment with L-carnitine. J Cardiovasc Pharmacol Ther 17:215–222. https://doi.org/10.1177/1074248411419502
Law A, Gauthier S, Quirion R (2001) Say NO to Alzheimer’s disease: the putative links between nitric oxide and dementia of the Alzheimer’s type. Brain Res Brain Res Rev 35:73–96. https://doi.org/10.1016/s0165-0173(00)00051-5
Liepinsh E, Vilskersts R, Skapare E, Svalbe B, Kuka J, Cirule H, Pugovics O, Kalvinsh I, Dambrova M (2008) Mildronate decreases carnitine availability and up-regulates glucose uptake and related gene expression in the mouse heart. Life Sci 83:613–619. https://doi.org/10.1016/j.lfs.2008.08.008
Low PA, Nickander KK, Tritschler HJ (1997) The roles of oxidative stress and antioxidant treatment in experimental diabetic neuropathy. Diabetes 46:S38–S42. https://doi.org/10.2337/diab.46.2.s38
Makrecka M, Svalbe B, Volska K, Sevostjanovs E, Liepins J, Grinberga S, Pugovics O, Liepinsh E, Dambrova M (2014) Mildronate, the inhibitor of L-carnitine transport, induces brain mitochondrial uncoupling and protects against anoxia-reoxygenation. Eur J Pharmacol 723:55–61. https://doi.org/10.1016/j.ejphar.2013.12.006
Mancuso C, Bates TE, Butterfield DA, Calafato S, Cornelius C, Lorenzo AD, Dinkova-Kostova AT, Calabrese V (2007) Natural antioxidants in Alzheimer’s disease. Expert Opin Investig Drugs 6:1921–1931. https://doi.org/10.1517/13543784.16.12.1921
Matthews BR (2015) Memory dysfunction. Continuum (Minneap Minn). 21:613–26. https://doi.org/10.1212/01.CON.0000466656.59413.29
Meeusen R, Decroix L (2018) Nutritional supplements and the brain. Int J Sport Nutr Exerc Metab 28:200–211. https://doi.org/10.1123/ijsnem.2017-0314
Muhammad T, Ikram M, Ullah R, Rehman SU, Kim MO (2019) Hesperetin, a citrus flavonoid, attenuates LPS-induced neuroinflammation, apoptosis and memory impairments by modulating TLR4/NF-κB signaling. Nutrients 11:648. https://doi.org/10.3390/nu11030648
Patching SG (2017) Glucose transporters at the blood-brain barrier: function, regulation and gateways for drug delivery. Mol Neurobiol 54:1046–1077. https://doi.org/10.1007/s12035-015-9672-6
Pourbagher-Shahri AM, Farkhondeh T, Talebi M, Kopustinskiene DM, Samarghandian S, Bernatoniene J (2021) An overview of NO signaling pathways in aging. Molecules 26:4533. https://doi.org/10.3390/molecules26154533
Pupure J, Fernandes MA, Santos MS, Moreno AJ, Kalvinsh I, Klusa V, Oliveira CR (2008) Mitochondria as the target for mildronate’s protective effects in azidothymidine (AZT)-induced toxicity of isolated rat liver mitochondria. Cell Biochem Funct 26:620–631. https://doi.org/10.1002/cbf.1486
Sakvarelidze EP, Zakaraia MV, Megreladze TI (2006) Change of concentration endothelin-1 in rats on a background of alcohol and/or mildronate treatment. Georgian Med News 135:131–134
Seeley JJ, Ghosh S (2017) Molecular mechanisms of innate memory and tolerance to LPS. J Leukoc Biol 101:107–119. https://doi.org/10.1189/jlb.3MR0316-118RR
Shaforostova EA, Gureev AP, Vitkalova IY, Popov VN (2021) The effect of L-carnitine depletion induced by long-term therapy of mice with meldonium on brain mitochondrial balance. Biomed Khim 67:74–80. https://doi.org/10.18097/PBMC20216701074
Sjakste N, Gutcaits A, Kalvinsh I (2005) Mildronate: an antiischemic drug for neurological indications CNS Drug Rev 11:151–68. https://doi.org/10.1111/j.1527-3458.2005.tb00267.x
Skelly DT, Griffin ÉW, Murray CL, Harney S, O’Boyle C, Hennessy E, Dansereau MA, Nazmi A, Tortorelli L, Rawlins JN, Bannerman DM, Cunningham C (2019) Acute transient cognitive dysfunction and acute brain injury induced by systemic inflammation occur by dissociable IL-1-dependent mechanisms. Mol Psychiatry 24:1533–1548. https://doi.org/10.1038/s41380-018-0075-8
Sorrenti V, Contarini G, Sut S, Dall’Acqua S, Confortin F, Pagetta A, Giusti P, Zusso M (2018) Curcumin prevents acute neuroinflammation and long-term memory impairment induced by systemic lipopolysaccharide in mice. Front Pharmacol 9:183. https://doi.org/10.3389/fphar.2018.00183
Steinmetz AB, Johnson SA, Iannitelli DE, Pollonini G, Alberini CM (2016) Insulin-like growth factor 2 rescues aging-related memory loss in rats. Neurobiol Aging 44:9–21. https://doi.org/10.1016/j.neurobiolaging.2016.04.006
Tang SS, Wang XY, Hong H, Long Y, Li YQ, Xiang GQ, Jiang LY, Zhang HT, Liu LP, Miao MX, Hu M, Zhang TT, Hu W, Ji H, Ye FY (2013) Leukotriene D4 induces cognitive impairment through enhancement of CysLT1R-mediated amyloid-β generation in mice. Neuropharmacology 65:182–192. https://doi.org/10.1016/j.neuropharm.2012.08.026
Traina G (2016) The neurobiology of acetyl-L-carnitine. Front Biosci (Landmark Ed) 21:1314–1329. https://doi.org/10.2741/4459
Tucker LB, Velosky AG, McCabe JT (2018) Applications of the Morris water maze in translational traumatic brain injury research. Neurosci Biobehav Rev 88:187–200. https://doi.org/10.1016/j.neubiorev.2018.03.010
Vorhees CV, Williams MT (2006) Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc 1:848–858. https://doi.org/10.1038/nprot.2006.116
Wang X, Bai Y, Cheng G, Ihsan A, Zhu F, Wang Y, Tao Y, Chen D, Dai M, Liu Z, Yuan Z (2016) Genomic and proteomic analysis of the inhibition of synthesis and secretion of aldosterone hormone induced by quinocetone in NCI-H295R cells. Toxicology 350–352:1–14. https://doi.org/10.1016/j.tox.2016.03.005
Winiarska-Mieczan A, Baranowska-Wójcik E, Kwiecień M, Grela ER, Szwajgier D, Kwiatkowska K, Kiczorowska B (2020) The role of dietary antioxidants in the pathogenesis of neurodegenerative diseases and their impact on cerebral oxidoreductive balance. Nutrients 12:435. https://doi.org/10.3390/nu12020435
Zakaria R, Wan Yaacob WMH, Othman Z, Long I, Ahmad AH, Al-Rahbi B (2017) Lipopolysaccharide-induced memory impairment in rats: a model of Alzheimer’s disease. Physiol Res 66:553–565. https://doi.org/10.33549/physiolres.933480
Zhang DM, Guo ZX, Zhao YL, Wang QJ, Gao YS, Yu T, Chen YK, Chen XM, Wang GQ (2019) L-carnitine regulated Nrf2/Keap1 activation in vitro and in vivo and protected oxidized fish oil-induced inflammation response by inhibiting the NF-κB signaling pathway in Rhynchocypris lagowski Dybowski. Fish Shellfish Immunol 93:1100–1110. https://doi.org/10.1016/j.fsi.2019.08.041
Zhu Y, Zhang G, Zhao J, Li D, Yan X, Liu J, Zhao H, Xia J, Zhang X, Li Z, Zhang B, Guo Z, Feng L, Zhang Z, Qu F, Zhao G (2013) Efficacy and safety of mildronate for acute ischemic stroke: a randomized, double-blind, active-controlled phase II multicenter trial. Clin Drug Investig 33:755–760. https://doi.org/10.1007/s40261-013-0121-x
Zhuo M, Hawkins RD (1995) Long-term depression: a learning-related type of synaptic plasticity in the mammalian central nervous system. Rev Neurosci 6:259–277. https://doi.org/10.1515/revneuro.1995.6.3.259
Zvejniece L, Svalbe B, Makrecka M, Liepinsh E, Kalvinsh I, Dambrova M (2010) Mildronate exerts acute anticonvulsant and antihypnotic effects. Behav Pharmacol 21:548–555. https://doi.org/10.1097/FBP.0b013e32833d5a59
Funding
The reported study was funded by RFBR, project number 19-34-90110, President grant for support of leading scientific school (Agreement NSh 1375.2022.5), by RF Ministry of Science and Higher Education in the framework of the national project “Science” (Agreement 075-03-2020-088, Unique number of the register of State tasks 075001 × 39782002).
Author information
Authors and Affiliations
Contributions
Conceptualization, A.P.G. and E.A.S.; Validation, E.A.S. and D.E.V.; Investigation, E.A.S. and D.E.V.; Writing—original draft preparation, E.A.S. and A.P.G.; Project administration, A.P.G.; Supervision, V.N.P.; Funding acquisition, V.N.P. All authors have read and agreed to the published version of the manuscript.
Corresponding author
Ethics declarations
Institutional review board statement
Animals used in the experiments were raised, kept, and sacrificed in accordance with the guidelines established by the Committee for the Care and Use of Animals of Voronezh State University, which comply with the Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the n the protection of animals used for scientific purposes.
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
ESM 1
(PDF 251 KB)
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Shaforostova, E.A., Gureev, A.P., Volodina, D.E. et al. Neuroprotective effect of mildronate and L-carnitine on the cognitive parameters of aged mice and mice with LPS-induced inflammation. Metab Brain Dis 37, 2497–2510 (2022). https://doi.org/10.1007/s11011-022-01047-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11011-022-01047-9