Liver Metabolomics and Inflammatory Profiles in Mouse Model of Fentanyl Overdose Treated with Beta-Lactams

Abstract

Fentanyl is a highly potent opioid analgesic that is approved medically to treat acute and chronic pain. There is a high potential for overdose-induced organ toxicities, including liver toxicity, and this might be due to the increase of recreational use of opioids. Several preclinical studies have demonstrated the efficacy of beta-lactams in modulating the expression of glutamate transporter-1 (GLT-1) in different body organs, including the liver. The upregulation of GLT-1 by beta-lactams is associated with the attenuation of hyperglutamatergic state, which is a characteristic feature of opioid use disorders. A novel experimental beta-lactam compound with no antimicrobial properties, MC-100093, has been developed to attenuate dysregulation of glutamate transport, in part by normalizing GLT-1 expression. A previous study showed that MC-100093 modulated hepatic GLT-1 expression with subsequent attenuation of alcohol-increased fat droplet content in the liver. In this study, we investigated the effects of fentanyl overdose on liver metabolites, and determined the effects of MC-100093 and ceftriaxone in the liver of a fentanyl overdose mouse model. Liver samples from control, fentanyl overdose, and fentanyl overdose ceftriaxone- or MC-100093-treated mice were analyzed for metabolomics using gas chromatography–mass spectrometry. Heatmap analysis revealed that both MC-100093 and ceftriaxone attenuated the effects of fentanyl overdose on several metabolites, and MC-100093 showed superior effects. Statistical analysis showed that MC-100093 reversed the effects of fentanyl overdose in some metabolites. Moreover, enrichment analysis revealed that the altered metabolites were strongly linked to the glucose-alanine cycle, the Warburg effect, gluconeogenesis, glutamate metabolism, lactose degradation, and ketone body metabolism. The changes in liver metabolites induced by fentanyl overdose were associated with liver inflammation, an effect attenuated with ceftriaxone pre-treatments. Ceftriaxone normalized fentanyl-overdose-induced changes in liver interleukin-6 and cytochrome CYP3A11 (mouse homolog of human CYP3A4) expression. Our data indicate that fentanyl overdose impaired liver metabolites, and MC-100093 restored certain metabolites.

Tackling new psychoactive substances through metabolomics: UHPLC-HRMS study on natural and synthetic opioids in male and female murine models

“…Up-regulation of glycolysis would lead to enhanced transamination of pyruvate to alanine, justifying the high levels of this amino acid in treated animals. A recent study by Alasmari et al 36 , in mouse model of fentanyl overdose (50 µg/kg i.p) suggested the involvement of glucose-alanine cycle and gluconeogenesis in liver metabolism alterations associated to increased liver inflammation, after a treatment with a fatal dose of fentanyl. These findings suggest that the increasing amino acids in urine samples of mice treated with fentanyl and morphine could be related to a protective response against increased inflammations triggered by these opioids in different organs 37 .…”

Effects of novel beta-lactam, MC-100093, and ceftriaxone on astrocytic glutamate transporters and neuroinflammatory factors in nucleus accumbens of C57BL/6 mice exposed to escalated doses of morphine

“…In addition, exposure of hippocampal cultures to morphine induced increases in neuroinflammatory cytokines, including IL-6 ( Osmanlioglu et al, 2020 ). Alternatively, a recent study from our group revealed that exposure to repeated exposures to fentanyl moderate dose followed by higher dose of fentanyl induced increases in IL-6 in liver, and this effect was attenuated by ceftriaxone treatment ( Alasmari et al, 2023 ). In the present study, both MC-100093 and ceftriaxone attenuated morphine-induced increase in IL-6 mRNA expression in the NAc.…”