Abstract
Synthetic cathinones are among the most popular new psychoactive substances, being abused for their stimulant properties, which are similar to those of amphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Considering that the liver is a likely target for cathinones-induced toxicity, and for their metabolic activation/detoxification, we aimed to determine the hepatotoxicity of three commonly abused synthetic cathinones: butylone, α-methylamino-butyrophenone (buphedrone) and 3,4-dimethylmethcathinone (3,4-DMMC). We characterized their cytotoxic profile in primary rat hepatocytes (PRH) and in the HepaRG and HepG2 cell lines. PRH was the most sensitive cell model, showing the lowest EC50 values for all three substances (0.158 mM for 3,4-DMMC; 1.21 mM for butylone; 1.57 mM for buphedrone). Co-exposure of PRH to the synthetic cathinones and CYP450 inhibitors (selective and non-selective) proved that hepatic metabolism reduced the toxicity of buphedrone but increased that of butylone and 3,4-DMMC. All compounds were able to increase oxidative stress, disrupting mitochondrial homeostasis and inducing apoptotic and necrotic features, while also increasing the occurrence of acidic vesicular organelles in PRH, compatible with autophagic activation. In conclusion, butylone, buphedrone and 3,4-DMMC have hepatotoxic potential, and their toxicity lies in the interference with a number of homeostatic processes, while being influenced by their metabolic fate.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adamowicz P, Gieron J, Gil D, Lechowicz W, Skulska A, Tokarczyk B (2016) The prevalence of new psychoactive substances in biological material—a three-year review of casework in Poland. Drug Test Anal 8(1):63–70. https://doi.org/10.1002/dta.1924
Aninat C, Piton A, Glaise D et al (2006) Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells. Drug Metab Dispos 34(1):75–83. https://doi.org/10.1124/dmd.105.006759
Araujo AM, Valente MJ, Carvalho M et al (2015) Raising awareness of new psychoactive substances: chemical analysis and in vitro toxicity screening of “legal high” packages containing synthetic cathinones. Arch Toxicol 89(5):757–771. https://doi.org/10.1007/s00204-014-1278-7
Arbo MD, Silva R, Barbosa DJ et al (2016) In vitro neurotoxicity evaluation of piperazine designer drugs in differentiated human neuroblastoma SH-SY5Y cells. J Appl Toxicol 36(1):121–130. https://doi.org/10.1002/jat.3153
Astashkina A, Mann B, Grainger DW (2012) A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity. Pharmacol Ther 134(1):82–106. https://doi.org/10.1016/j.pharmthera.2012.01.001
Bachour-El Azzi P, Sharanek A, Burban A et al (2015) Comparative localization and functional activity of the main hepatobiliary transporters in HepaRG cells and primary human hepatocytes. Toxicol Sci 145(1):157–168. https://doi.org/10.1093/toxsci/kfv041
Beitia G, Cobreros A, Sainz L, Cenarruzabeitia E (1999) 3,4-Methylenedioxymethamphetamine (ecstasy)-induced hepatotoxicity: effect on cytosolic calcium signals in isolated hepatocytes. Liver 19(3):234–241
Bouma ME, Rogier E, Verthier N, Labarre C, Feldmann G (1989) Further cellular investigation of the human hepatoblastoma-derived cell line HepG2: morphology and immunocytochemical studies of hepatic-secreted proteins. Vitro Cell Dev Biol 25(3 Pt 1):267–275
Calinski DM, Kisor DF, Sprague JE (2018) A review of the influence of functional group modifications to the core scaffold of synthetic cathinones on drug pharmacokinetics. Psychopharmacology 236(3):881–890. https://doi.org/10.1007/s00213-018-4985-6
Cao L, Quan XB, Zeng WJ, Yang XO, Wang MJ (2016) Mechanism of hepatocyte apoptosis. J Cell Death 9:19–29. https://doi.org/10.4137/JCD.S39824
Carvalho F, Remiao F, Amado F, Domingues P, Correia AJ, Bastos ML (1996) d-Amphetamine interaction with glutathione in freshly isolated rat hepatocytes. Chem Res Toxicol 9(6):1031–1036. https://doi.org/10.1021/tx9501750
Carvalho M, Carvalho F, Bastos ML (2001) Is hyperthermia the triggering factor for hepatotoxicity induced by 3,4-methylenedioxymethamphetamine (ecstasy)? An in vitro study using freshly isolated mouse hepatocytes. Arch Toxicol 74(12):789–793
Carvalho M, Remiao F, Milhazes N et al (2004) The toxicity of N-methyl-alpha-methyldopamine to freshly isolated rat hepatocytes is prevented by ascorbic acid and N-acetylcysteine. Toxicology 200(2–3):193–203. https://doi.org/10.1016/j.tox.2004.03.016
Carvalho M, Carmo H, Costa VM et al (2012) Toxicity of amphetamines: an update. Arch Toxicol 86(8):1167–1231. https://doi.org/10.1007/s00204-012-0815-5
Cerretani D, Bello S, Cantatore S et al (2011) Acute administration of 3,4-methylenedioxymethamphetamine (MDMA) induces oxidative stress, lipoperoxidation and TNFalpha-mediated apoptosis in rat liver. Pharmacol Res 64(5):517–527. https://doi.org/10.1016/j.phrs.2011.08.002
Chandramani Shivalingappa P, Jin H, Anantharam V, Kanthasamy A, Kanthasamy A (2012) N-acetyl cysteine protects against methamphetamine-induced dopaminergic neurodegeneration via modulation of redox status and autophagy in dopaminergic cells. Parkinsons Dis 2012:424285. https://doi.org/10.1155/2012/424285
De Letter EA, Piette MH, Lambert WE, Cordonnier JA (2006) Amphetamines as potential inducers of fatalities: a review in the district of Ghent from 1976–2004. Med Sci Law 46(1):37–65. https://doi.org/10.1258/rsmmsl.46.1.37
Dias da Silva D, Carmo H, Lynch A, Silva E (2013a) An insight into the hepatocellular death induced by amphetamines, individually and in combination: the involvement of necrosis and apoptosis. Arch Toxicol 87(12):2165–2185. https://doi.org/10.1007/s00204-013-1082-9
Dias da Silva D, Silva E, Carmo H (2013b) Cytotoxic effects of amphetamine mixtures in primary hepatocytes are severely aggravated under hyperthermic conditions. Toxicol In Vitro 27(6):1670–1678. https://doi.org/10.1016/j.tiv.2013.04.010
Dias da Silva D, Silva E, Carmo H (2014a) Combination effects of amphetamines under hyperthermia—the role played by oxidative stress. J Appl Toxicol 34(6):637–650. https://doi.org/10.1002/jat.2889
Dias da Silva D, Silva E, Carvalho F, Carmo H (2014b) Mixtures of 3,4-methylenedioxymethamphetamine (ecstasy) and its major human metabolites act additively to induce significant toxicity to liver cells when combined at low, non-cytotoxic concentrations. J Appl Toxicol 34(6):618–627. https://doi.org/10.1002/jat.2885
Dias da Silva D, Arbo MD, Valente MJ, Bastos ML, Carmo H (2015) Hepatotoxicity of piperazine designer drugs: Comparison of different in vitro models. Toxicol In Vitro 29(5):987–996. https://doi.org/10.1016/j.tiv.2015.04.001
Dias da Silva D, Silva MJ, Moreira P et al (2017) In vitro hepatotoxicity of “Legal X”: the combination of 1-benzylpiperazine (BZP) and 1-(m-trifluoromethylphenyl)piperazine (TFMPP) triggers oxidative stress, mitochondrial impairment and apoptosis. Arch Toxicol 91(3):1413–1430. https://doi.org/10.1007/s00204-016-1777-9
Dias da Silva D, Ferreira B, Roque Bravo R et al (2019) The new psychoactive substance 3-methylmethcathinone (3-MMC or metaphedrone) induces oxidative stress, apoptosis, and autophagy in primary rat hepatocytes at human-relevant concentrations. Arch Toxicol 93(9):2617–2634. https://doi.org/10.1007/s00204-019-02539-x
Drug Enforcement Administration (2011) Schedules of controlled substances: temporary placement of three synthetic cathinones in Schedule I. Final Order. Fed Regist 76(204):65371–65375
El-Tawil OS, Abou-Hadeed AH, El-Bab MF, Shalaby AA (2011) d-Amphetamine-induced cytotoxicity and oxidative stress in isolated rat hepatocytes. Pathophysiology 18(4):279–285. https://doi.org/10.1016/j.pathophys.2011.04.001
EMCDDA (2020) European Drug Report 2020: trends and development. European Monitoring Centre for Drugs and Drug Addiction, Luxembourg
EMCDDA-Europol (2011) EMCDDA–Europol 2010 Annual Report on the implementation of Council Decision 2005/387/JHA, Lisbon
Faber KN, Muller M, Jansen PL (2003) Drug transport proteins in the liver. Adv Drug Deliv Rev 55(1):107–124
Forman HJ, Zhang H, Rinna A (2009) Glutathione: overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med 30(1–2):1–12. https://doi.org/10.1016/j.mam.2008.08.006
Frohlich S, Lambe E, O’Dea J (2011) Acute liver failure following recreational use of psychotropic “head shop” compounds. Ir J Med Sci 180(1):263–264. https://doi.org/10.1007/s11845-010-0636-6
Garcia-Repetto R, Moreno E, Soriano T, Jurado C, Gimenez MP, Menendez M (2003) Tissue concentrations of MDMA and its metabolite MDA in three fatal cases of overdose. Forensic Sci Int 135(2):110–114
Gaspar H, Bronze S, Oliveira C et al (2018) Proactive response to tackle the threat of emerging drugs: Synthesis and toxicity evaluation of new cathinones. Forensic Sci Int 290:146–156. https://doi.org/10.1016/j.forsciint.2018.07.001
Gerets HH, Tilmant K, Gerin B et al (2012) Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol Toxicol 28(2):69–87. https://doi.org/10.1007/s10565-011-9208-4
Glicksberg L, Bryand K, Kerrigan S (2016) Identification and quantification of synthetic cathinones in blood and urine using liquid chromatography-quadrupole/time of flight (LC-Q/TOF) mass spectrometry. J Chromatogr B Anal Technol Biomed Life Sci 1035:91–103. https://doi.org/10.1016/j.jchromb.2016.09.027
Green CE, LeValley SE, Tyson CA (1986) Comparison of amphetamine metabolism using isolated hepatocytes from five species including human. J Pharmacol Exp Ther 237(3):931–936
Guillouzo A, Corlu A, Aninat C, Glaise D, Morel F, Guguen-Guillouzo C (2007) The human hepatoma HepaRG cells: a highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. Chem Biol Interact 168(1):66–73. https://doi.org/10.1016/j.cbi.2006.12.003
Helander A, Backberg M, Hulten P, Al-Saffar Y, Beck O (2014) Detection of new psychoactive substance use among emergency room patients: results from the Swedish STRIDA project. Forensic Sci Int 243:23–29. https://doi.org/10.1016/j.forsciint.2014.02.022
Hilgendorf C, Ahlin G, Seithel A, Artursson P, Ungell AL, Karlsson J (2007) Expression of thirty-six drug transporter genes in human intestine, liver, kidney, and organotypic cell lines. Drug Metab Dispos 35(8):1333–1340. https://doi.org/10.1124/dmd.107.014902
Hiramatsu M, Kumagai Y, Unger SE, Cho AK (1990) Metabolism of methylenedioxymethamphetamine: formation of dihydroxymethamphetamine and a quinone identified as its glutathione adduct. J Pharmacol Exp Ther 254(2):521–527
Köppe H, Ludwig G, Zeile K (1967) Verfahren zur Herstellung von substituierten Phenyl-alpha-aminoketonen und deren Säureadditionssalzen bzw. deren optischen Antipoden. Germany Patent DE1242241
Larsen KE, Fon EA, Hastings TG, Edwards RH, Sulzer D (2002) Methamphetamine-induced degeneration of dopaminergic neurons involves autophagy and upregulation of dopamine synthesis. J Neurosci 22(20):8951–8960
Le Vee M, Jigorel E, Glaise D, Gripon P, Guguen-Guillouzo C, Fardel O (2006) Functional expression of sinusoidal and canalicular hepatic drug transporters in the differentiated human hepatoma HepaRG cell line. Eur J Pharm Sci 28(1–2):109–117. https://doi.org/10.1016/j.ejps.2006.01.004
Le Vee M, Noel G, Jouan E, Stieger B, Fardel O (2013) Polarized expression of drug transporters in differentiated human hepatoma HepaRG cells. Toxicol In Vitro 27(6):1979–1986. https://doi.org/10.1016/j.tiv.2013.07.003
Lee J, Giordano S, Zhang J (2012) Autophagy, mitochondria and oxidative stress: cross-talk and redox signalling. Biochem J 441(2):523–540. https://doi.org/10.1042/BJ20111451
Li IH, Ma KH, Weng SJ, Huang SS, Liang CM, Huang YS (2014) Autophagy activation is involved in 3,4-methylenedioxymethamphetamine ('ecstasy’)-induced neurotoxicity in cultured cortical neurons. PLoS ONE 9(12):e116565. https://doi.org/10.1371/journal.pone.0116565
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275
Luethi D, Liechti ME, Krahenbuhl S (2017) Mechanisms of hepatocellular toxicity associated with new psychoactive synthetic cathinones. Toxicology 387:57–66. https://doi.org/10.1016/j.tox.2017.06.004
Luethi D, Kolaczynska KE, Docci L, Krahenbuhl S, Hoener MC, Liechti ME (2018) Pharmacological profile of mephedrone analogs and related new psychoactive substances. Neuropharmacology 134(Pt A):4–12. https://doi.org/10.1016/j.neuropharm.2017.07.026
Luethi D, Walter M, Zhou X, Rudin D, Krahenbuhl S, Liechti ME (2019) Para-halogenation affects monoamine transporter inhibition properties and hepatocellular toxicity of amphetamines and methcathinones. Front Pharmacol 10:438. https://doi.org/10.3389/fphar.2019.00438
Martins MJ, Roque Bravo R, Enea M et al (2018) Ethanol addictively enhances the in vitro cardiotoxicity of cocaine through oxidative damage, energetic deregulation, and apoptosis. Arch Toxicol 92(7):2311–2325. https://doi.org/10.1007/s00204-018-2227-7
Mayer FP, Schmid D, Owens WA et al (2018) An unsuspected role for organic cation transporter 3 in the actions of amphetamine. Neuropsychopharmacology 43(12):2408–2417. https://doi.org/10.1038/s41386-018-0053-5
Mayer FP, Schmid D, Holy M, Daws LC, Sitte HH (2019) “Polytox” synthetic cathinone abuse: a potential role for organic cation transporter 3 in combined cathinone-induced efflux. Neurochem Int 123:7–12. https://doi.org/10.1016/j.neuint.2018.09.008
Mercer LD, Higgins GC, Lau CL, Lawrence AJ, Beart PM (2017) MDMA-induced neurotoxicity of serotonin neurons involves autophagy and rilmenidine is protective against its pathobiology. Neurochem Int 105:80–90. https://doi.org/10.1016/j.neuint.2017.01.010
Montiel-Duarte C, Varela-Rey M, Oses-Prieto JA et al (2002) 3,4-Methylenedioxymethamphetamine (“Ecstasy”) induces apoptosis of cultured rat liver cells. Biochim Biophys Acta 1588(1):26–32
Mueller DM, Rentsch KM (2012) Generation of metabolites by an automated online metabolism method using human liver microsomes with subsequent identification by LC-MS(n), and metabolism of 11 cathinones. Anal Bioanal Chem 402(6):2141–2151. https://doi.org/10.1007/s00216-011-5678-8
Nakagawa Y, Suzuki T, Tayama S, Ishii H, Ogata A (2009) Cytotoxic effects of 3,4-methylenedioxy-N-alkylamphetamines, MDMA and its analogues, on isolated rat hepatocytes. Arch Toxicol 83(1):69–80. https://doi.org/10.1007/s00204-008-0323-9
Nikoletopoulou V, Markaki M, Palikaras K, Tavernarakis N (2013) Crosstalk between apoptosis, necrosis and autophagy. Biochim Biophys Acta 1833(12):3448–3459. https://doi.org/10.1016/j.bbamcr.2013.06.001
Pedersen AJ, Petersen TH, Linnet K (2013a) In vitro metabolism and pharmacokinetic studies on methylone. Drug Metab Dispos 41(6):1247–1255. https://doi.org/10.1124/dmd.112.050880
Pedersen AJ, Reitzel LA, Johansen SS, Linnet K (2013b) In vitro metabolism studies on mephedrone and analysis of forensic cases. Drug Test Anal 5(6):430–438. https://doi.org/10.1002/dta.1369
Perry SW, Norman JP, Barbieri J, Brown EB, Gelbard HA (2011) Mitochondrial membrane potential probes and the proton gradient: a practical usage guide. Biotechniques 50(2):98–115. https://doi.org/10.2144/000113610
Pontes H, Duarte JA, de Pinho PG et al (2008) Chronic exposure to ethanol exacerbates MDMA-induced hyperthermia and exposes liver to severe MDMA-induced toxicity in CD1 mice. Toxicology 252(1–3):64–71. https://doi.org/10.1016/j.tox.2008.07.064
Richter LHJ, Flockerzi V, Maurer HH, Meyer MR (2017) Pooled human liver preparations, HepaRG, or HepG2 cell lines for metabolism studies of new psychoactive substances? A study using MDMA, MDBD, butylone, MDPPP, MDPV, MDPB, 5-MAPB, and 5-API as examples. J Pharm Biomed Anal 143:32–42. https://doi.org/10.1016/j.jpba.2017.05.028
Richter LHJ, Beck A, Flockerzi V, Maurer HH, Meyer MR (2019) Cytotoxicity of new psychoactive substances and other drugs of abuse studied in human HepG2 cells using an adopted high content screening assay. Toxicol Lett 301:79–89. https://doi.org/10.1016/j.toxlet.2018.11.007
Rodrigues RM, Bouhifd M, Bories G et al (2013) Assessment of an automated in vitro basal cytotoxicity test system based on metabolically-competent cells. Toxicol In Vitro 27(2):760–767. https://doi.org/10.1016/j.tiv.2012.12.004
Rojek S, Klys M, Strona M, Maciow M, Kula K (2012) “Legal highs”–toxicity in the clinical and medico-legal aspect as exemplified by suicide with bk-MBDB administration. Forensic Sci Int 222(1–3):e1-6. https://doi.org/10.1016/j.forsciint.2012.04.034
Roque Bravo R, Carmo H, Silva JP et al (2019) Emerging club drugs: 5-(2-aminopropyl)benzofuran (5-APB) is more toxic than its isomer 6-(2-aminopropyl)benzofuran (6-APB) in hepatocyte cellular models. Arch Toxicol. https://doi.org/10.1007/s00204-019-02638-9
Rouxinol D, Carmo H, Carvalho F, Bastos MdL, Dias da Silva D (2020a) Pharmacokinetics, pharmacodynamics, and toxicity of the new psychoactive substance 3,4-dimethylmethcathinone (3,4-DMMC). Forensic Toxicol 38(1):15–29. https://doi.org/10.1007/s11419-019-00494-x
Rouxinol D, Dias da Silva D, Silva JP, Carvalho F, Bastos ML, Carmo H (2020b) Biodistribution and metabolic profile of 3,4-dimethylmethcathinone (3,4-DMMC) in Wistar rats through gas chromatography-mass spectrometry (GC-MS) analysis. Toxicol Lett 320:113–123. https://doi.org/10.1016/j.toxlet.2019.10.003
Shima N, Katagi M, Kamata H et al (2013) Urinary excretion and metabolism of the newly encountered designer drug 3,4-dimethylmethcathinone in humans. Forensic Toxicol 31(1):101–112. https://doi.org/10.1007/s11419-012-0172-3
Simmler LD, Buser TA, Donzelli M et al (2013) Pharmacological characterization of designer cathinones in vitro. Br J Pharmacol 168(2):458–470. https://doi.org/10.1111/j.1476-5381.2012.02145.x
Simmler LD, Rickli A, Hoener MC, Liechti ME (2014) Monoamine transporter and receptor interaction profiles of a new series of designer cathinones. Neuropharmacology 79:152–160. https://doi.org/10.1016/j.neuropharm.2013.11.008
Sykutera M, Cychowska M, Bloch-Boguslawska E (2015) A fatal case of pentedrone and alpha-pyrrolidinovalerophenone poisoning. J Anal Toxicol 39(4):324–329. https://doi.org/10.1093/jat/bkv011
Uralets V, Rana S, Morgan S, Ross W (2014) Testing for designer stimulants: metabolic profiles of 16 synthetic cathinones excreted free in human urine. J Anal Toxicol 38(5):233–241. https://doi.org/10.1093/jat/bku021
Usui K, Aramaki T, Hashiyada M, Hayashizaki Y, Funayama M (2014) Quantitative analysis of 3,4-dimethylmethcathinone in blood and urine by liquid chromatography-tandem mass spectrometry in a fatal case. Leg Med (Tokyo) 16(4):222–226. https://doi.org/10.1016/j.legalmed.2014.03.008
Valente MJ, Araujo AM, Silva R et al (2015) 3,4-Methylenedioxypyrovalerone (MDPV): in vitro mechanisms of hepatotoxicity under normothermic and hyperthermic conditions. Arch Toxicol 90(8):1959–1973. https://doi.org/10.1007/s00204-015-1653-z
Valente MJ, Araujo AM, Bastos ML et al (2016) Characterization of hepatotoxicity mechanisms triggered by designer cathinone drugs (beta-keto amphetamines). Toxicol Sci 153(1):89–102. https://doi.org/10.1093/toxsci/kfw105
Valente MJ, Amaral C, Correia-da-Silva G et al (2017a) Methylone and MDPV activate autophagy in human dopaminergic SH-SY5Y cells: a new insight into the context of beta-keto amphetamines-related neurotoxicity. Arch Toxicol 91(11):3663–3676. https://doi.org/10.1007/s00204-017-1984-z
Valente MJ, Bastos ML, Fernandes E, Carvalho F, Guedes de Pinho P, Carvalho M (2017b) Neurotoxicity of beta-keto amphetamines: deathly mechanisms elicited by methylone and MDPV in human dopaminergic SH-SY5Y Cells. ACS Chem Neurosci 8(4):850–859. https://doi.org/10.1021/acschemneuro.6b00421
Warrick BJ, Wilson J, Hedge M, Freeman S, Leonard K, Aaron C (2012) Lethal serotonin syndrome after methylone and butylone ingestion. J Med Toxicol 8(1):65–68. https://doi.org/10.1007/s13181-011-0199-6
Yokchue T, Anderson R (2015) In vitro metabolism studies on methylenedioxy-substituted amphetamines using human liver microsomes and LC/MS/MS with chemical derivatization. In: International association of forensic toxicologist meeting, Atlanta, GA, USA
Yuan L, Kaplowitz N (2013) Mechanisms of drug-induced liver injury. Clin Liver Dis 17(4):507–518. https://doi.org/10.1016/j.cld.2013.07.002
Zaitsu K, Katagi M, Kamata HT et al (2009) Determination of the metabolites of the new designer drugs bk-MBDB and bk-MDEA in human urine. Forensic Sci Int 188(1–3):131–139. https://doi.org/10.1016/j.forsciint.2009.04.001
Zhang Y, Chen X, Gueydan C, Han J (2018) Plasma membrane changes during programmed cell deaths. Cell Res 28(1):9–21. https://doi.org/10.1038/cr.2017.133
Zuba D, Adamowicz P, Byrska B (2013) Detection of buphedrone in biological and non-biological material—two case reports. Forensic Sci Int 227(1–3):15–20. https://doi.org/10.1016/j.forsciint.2012.08.034
Funding
This work was financed by FEDER funds through the COMPETE 2020—Operational Programme for Competitiveness and Internationalisation (POCI), and by Portuguese funds through FCT (Fundação para a Ciência e a Tecnologia) in the framework of the project POCI-01-0145-FEDER-029584. This work was also supported by the Applied Molecular Biosciences Unit—UCIBIO which is financed by national funds from FCT (UIDP/04378/2020 and UIDB/04378/2020).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declared that they have no conflicts of interest
Ethics approval
All procedures involving animals were executed considering the highest standards of ethics, taking into consideration international, national and/or institutional guidelines. Procedures were performed after approval by the local Ethical Committee for the Welfare of Experimental Animals (University of Porto-ORBEA; project 158/2014) and by the national authority Direção Geral de Alimentação e Veterinária (DGAV).
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Roque Bravo, R., Carmo, H., Valente, M.J. et al. From street to lab: in vitro hepatotoxicity of buphedrone, butylone and 3,4-DMMC. Arch Toxicol 95, 1443–1462 (2021). https://doi.org/10.1007/s00204-021-02990-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-021-02990-9