Glycosphingolipids Prevent APAP and HMG-CoA Reductase Inhibitors-mediated Liver Damage: A Novel Method for “Safer Drug” Formulation that Prevents Drug-induced Liver Injury

Abstract

Background and Aims: Acetaminophen (APAP) and HMG-CoA reductase inhibitors are common causes of drug-induced liver injury (DILI). This study aimed to determine the ability to reduce APAP- and statins-mediated liver injury by using formulations that combine glycosphingolipids and vitamin E.Methods: Mice were injected with APAP or with statins and treated before and after with β-glucosylceramide (GC), with or without vitamin E. Mice were followed for changes in liver enzymes, liver histology, hepatic expression of JNK, STAT3 and caspase 3, as well as intrahepatic natural killer T cells (NKT) and the serum cytokine levels by flow cytometry.Results: Administration of GC before or after APAP alleviated the liver damage, as noted by a reduction of the liver enzymes, improvement in the liver histology and decreased hepatic caspase 3 expression. Beneficial effect was associated with a reduction of the intrahepatic NKT, JNK expression in the liver, and increased glutathione in the liver, and decreased TNF-α serum levels. Synergistic effect of co-administration of GC with vitamin E was observed. Similar protective effect of GC on statin-mediated liver damage was documented by a reduction in liver enzymes and improved liver histology, which was mediated by reduction of NKT, increased STAT3 expression in the liver, and reduced the TGF-β and IL17 levels.Conclusions: β-glycosphingolipids exert a hepatoprotective effect on APAP- and statins-mediated liver damage. Vitamin E exerted a synergistic effect to that of GC. The generation of “safer drug” formulations, which include an active molecule combined with a hepatoprotective adjuvant, may provide an answer to the real unmet need of DILI.

Increased hepatic Akt phosphorylation alleviated glucose intolerance and improved liver function in leptin-deficient mice

“…The combination therapy of GC and insulin showed a synergistic effect for promoting Akt expression in the liver and alleviating glucose intolerance. An adjuvant effect of GC was previously described in the ob/ob model when co-administered with anti-CD3 [ 31 ] and with other drugs and vaccines [ 25 , 36 ]. These effects were attributed to the potential immunomodulatory effect of GSLs [ 19 , 37 ].…”

“…β-glucosylceramide was purchased from Avanti Polar Lipids (Alabaster, AL, USA), dissolved in a vehicle of a mixture of 30% Cremophor EL (Sigma, Rehovot,Israel) and ethanol (1 : 1) in PBS, and administered orally by gavage using a designated pipette at a dose of 2.5 mg/kg/day as described [ 21 , 23 , 25 , 31 ].…”

“…The number of animals was calculated based on previously published data [ 21 , 31 ]. Mice in all groups were followed for changes in metabolic parameters and parameters indicative of liver damage, as described [ 21 , 23 , 25 ]. The last treatment was administered the day before the end of the study.…”

“…β-glucosylceramide (GC) is a GSL shown to exert immunomodulatory effects [19][20][21][22][23][24][25][26][27]. Oral administration of GC alleviated insulin resistance in several animal models [20,21,28], and preliminary results suggest a beneficial effect on type 2 diabetes and non-alcoholic fatty liver disease (NAFLD) [29].…”

Immune-Mediated Drug-Induced Liver Injury: Immunogenetics and Experimental Models

“…For example, in halothane-DILI, a pro-inflammatory role of iNKT cells has been suggested, as a greatly reduced neutrophil infiltration was observed in CD1d −/− mice [ 45 ]. Similarly, in APAP-DILI, intrahepatic iNKT cell reduction or loss were associated to either ameliorated disease after treatment with glycosphingolipids and/or vitamin E [ 46 ], or increased glutathione levels and enhanced NAPQI (an APAP reactive metabolite) detoxification [ 47 ]. In contrast, another study showed exacerbated liver damage in both Jα18 −/− and CD1d −/− mice after starvation, and this was associated with CYP2E1 up-regulation and enhanced formation of hepatic APAP-protein adducts, suggesting a protective role of iNKT cells [ 48 ].…”