The impact of ademetionine and ipidacrine/phenibut on the NCAM distribution and behavior in the rat model of drug-induced liver injury

Introduction. Recently, more attention is being paid to the drug-induced liver injury (DILI) as a consequence of the tuberculos is treatment and the need for new medicine is emphasized. The use of isoniazid and rifampicin has a potentiating effect, which increases the risk of substancial liver damage. In turn, systemic accumulation of toxic metabolites leads to negative changes in various organs, including the brain. It causes an imbalance in biochemical and neurophysiological processes in the brain, ultimately giving the onset to the development of hepatic encephalopathy. Aim. The effects of rifampicin and isoniazid on the central nervous system have not been studied before and we aimed to evaluate the impact these two substances have on the neuronal cell adhesion molecules (NCAM) distribution and animal behavior in the rat model of DILI. Material and methods. The 24 male Wistar rats, weighing 180-220 g were used for the experiment and divided to the groups (n=6): 1 – control; 2 – rats with experimental DILI; 3 – rats with DILI plus the intravenous infusion of S-adenosyl-L-methionine at a dose of 35 mg/kg; 4 – rats with DILI plus a fixed combination of ipidacrine hydrochloride at a dose 1 mg/kg body weight and phenibut at a dose 60 mg/kg body weight daily for the last 14 days of the experiment. All experimental procedures were carried out in the accordance with the principles outlined in the current Guide to the Care and Use of Experimental Animals. The locomotor and research activities were studied in the open field test. The activity of aspartate aminotransferase (AST, ЕС 2.6.1.1) and alanine aminotransferase (ALT, ЕС 2.6.1.2) in the serum of rats were tested to confirm the liver damage. The quantitative analyses of soluble and membrane forms of NCAM were performed with ELISA. The ANOVA followed by a Tukey post-hoc test was used to assess statistical differences between groups. ORIGINAL PAPER Received: 1.06.2020 | Accepted: 6.07.2020 Publication date: September 2020 Participation of co-authors: A – Author of the concept and objectives of paper; B – collection of data; C – implementation of research; D – elaborate, analysis and interpretation of data; E – statistical analysis; F – preparation of a manuscript; G – working out the literature; H – obtaining funds; * Authors made an equal contribution © Wydawnictwo UR 2020 ISSN 2544-1361 (online); ISSN 2544-2406 doi: 10.15584/ejcem.2020.3.1 Corresponding author: Galyna Ushakova, e-mail: ushakovagalyna@gmail.com Muraviova D, Kharchenko Y, Pierzynowska K et al. The impact of ademetionine and ipidacrine/phenibut on the NCAM distribution and behavior in the rat model of drug-induced liver injury. Eur J Clin Exp Med. 2020;18(3):155–164. doi: 10.15584/ejcem.2020.3.1 Mariusz Wójcik http://orcid.org/0000-0002-1599-2394 Joanna Daszyk-Wójcik http://orcid.org/0000-0002-8819-5281 Kamil Skoczyński http://orcid.org/0000-0002-7836-0677 Daria Lahoda http://orcid.org/0000-0003-0783-6225 Valentyna Velychko Andressa Bonito Lopes http://orcid.org/0000-0002-3311-6363 Dhebora Espindola Amboni http://orcid.org/0000-0003-4794-3922 Marilis Macedo Schmidel http://orcid.org/0000-0002-7535-512 Miriélly Junges Maciel http://orcid.org/0000-0002-4582-5319 Alberito Rodrigo de Carvalho http://orcid.org/0000-0002-5520-441X Gladson Ricardo Flor Bertolini http://orcid.org/0000-0003-0565-2019 156 European Journal of Clinical and Experimental Medicine 2020; 18 (3):155–164 Introduction Drug-induced liver injury (DILI) accounts for up to 10 % of all adverse reactions associated with the use of drugs. According to World Health Organization (WHO), 50 out of 1000 patients are hospitalized due to the drug-induced complications.1,2 Information provided by WEB-platform LiverTox (http://livertox.nlm.nih. gov) indicates that 353 (53 %) of the 671 drugs available for analysis provoke the hepatotoxicity. Moreover, isoniazid, pyrazinamide, and rifampicin are the most toxic liver agents. In particular, in studies published in the United States in 2015, based on the analysis of more than 600 cases in 12 clinical trials, 46 % of DILI cases were associated with antimicrobial drugs, such as antibiotics and antitubercular drugs.2 The simultaneous use of isoniazid and rifampicin causes a potentiating effect, which increases the risk of substancial liver damage.3 The asymptomatic «subclinical» course of drug-induced hepatitis is dangerous, and further administration of these agents leads to the development of severe hepatitis, accompanied by jaundice and hepatic encephalopathy, which manifests as acognitive impairment.4,5 The main role in the development of these disorders be longs to the diminished detox function of the liver. Non-ionized ammonia easily penetrates the blood-brain barrier (BBB) and enters the astrocytes, where itismetabolized in the mitochondria in the presence of α-ketoglutarate to form of glutamine, a key component in the development of the astrocytic edema.6 The active forms of oxygen and nitrogen are highly reactive molecules and are redox-active compounds, which, depending on the concentration, have both a positive (proliferation of cells) and negative effects (cell growth arrest, cell death) on nerve cells.7,8 Oxidative and nitrosating stresses initiate neuro transmission disorders, mitochondrial dysfunction, and energy metabolism disorders in central nervous system (CNS).9,10 The cognitive deficits observed in hepatic encephalopathy are also the result of the synaptic plasticity violations and changes in mediator transmission.11 In particular, it is reported that hepatic encephalopathy is associated with highactivity of the GABA-ergic system of the brain due to increased γ-aminobutyric acid (GABA) concentration, expression of GABA receptors, and production of neurotrophic steroids, specifically aloprengonone.12 At the same time, there is a decrease in glutamatergic neurotransmission characteristic for the chronic liver disorders, and observed violations in learning and memory may be associated with the inhibition of the glutamate nitrogen oxide-cGMP regulatory effect in the hippocampus which appearsas a response to hyperammonemia or increased levels of dopamine which result form the impaired liver function.13 Our study aimed to investigate neuronal cell adhesion molecules (NCAM) in various areas of the brain after the long-term administration of rifampicin and isoniazidin rats. NCAMs play an important roles in the regulation of neuronal differentiation and migration by interacting with growth factors and their receptors, as well as in the mechanisms of membrane potential regulation, determining the excitability of neurons.14-16 Also, NCAMs in fluence the synaptic plasticity and cognitive processes of the mature brain, the short-term plasticity of existing synapses, and long-lasting plasticity associated with the elimination of old synapses and the formation of new ones. According to the contemporary notions about the functional role of NCAM, the blocked function of these proteins can lead to cognitive and emotional declines, such as changes in the perception of odor, memory, hearing, anxiety and space orientation.17 A lot of information on the effects of antimicrobials on the CNS has been collected over the last decade. However, the effect of rifampicin and isoniazid on the CNS could be felt no tonly by their expressive hepatotoxicity after prolonged use bu talso by the ability to cause a disturbance in the balance of the intestinal microflora and the development of dysbiosis, which potentially have negative impactas on the CNS it self as on the progression of toxic liver injury induced encephalopathy.18 Moreover, the membrane-bound proteins, including NCAM are recognized as the primar target for the endotoxins’negative effects on CNS.19 Therefore, the study of neuronal plasticity in the DILI model, as well as the possible ways of their pharmacological correction, is very relevant and could reveal new findings on the pathogenesis of cognitive impairmentin DILI. As a correction of pathological conditions, when using antitubercular therapy, various substances and preparations are tested. They have various mechanisms of action (direct and indirect) that are scantily studied. In our study, Results. Our investigation in the open field test revealed a significant decrease in the locomotor and research activity of rats after 28 days of rifampicin and isoniazid administration. The recovery of investigated parameters was observed in groups of animals treated with ademetionine (AD group) or combination of ipidacrine and phenibut (IP/PB group). We also observed that changes in rats’ behavior were consistent with alterations of the NCAM levels in the thalamus and hippocampus. Thus, the level of membrane NCAM was significantly decreased under DILI in both investigated brain regions (thalamus and hippocampus), while both AD and IP/PB treatments restored membrane NCAM levels towards those observed in the control group at least in the hippocampus. Conclusion. Obtained data suggests that both ademetionine and combinated drug containing ipidacrine and phenibut possesses neuroprotective properties and could prevent the decline in synaptic plasticity under antitubercular therapy.