Structural and Functional Interaction of Δ9-Tetrahydrocannabinol with Liver Fatty Acid Binding Protein (FABP1)

Abstract

Although serum Δ⁹-tetrahydrocannabinol (Δ⁹-THC) undergoes rapid hepatic clearance and metabolism, almost nothing is known regarding the mechanism(s) whereby this highly lipophilic phytocannabinoid is transported for metabolism/excretion. A novel NBD-arachidonoylethanolamide (NBD-AEA) fluorescence displacement assay showed that liver fatty acid binding protein (FABP1), the major hepatic endocannabinoid (EC) binding protein, binds the first major metabolite of Δ⁹-THC (Δ⁹-THC-OH) as well as Δ⁹-THC itself. Circular dichroism (CD) confirmed that not only Δ⁹-THC and Δ⁹-THC-OH but also downstream metabolites Δ⁹-THC-COOH and Δ⁹-THC-CO-glucuronide directly interact with FABP1. Δ⁹-THC and metabolite interaction differentially altered the FABP1 secondary structure, increasing total α-helix (all), decreasing total β-sheet (Δ⁹-THC-COOH, Δ⁹-THC-CO-glucuronide), increasing turns (Δ⁹-THC-OH, Δ⁹-THC-COOH, Δ⁹-THC-CO-glucuronide), and decreasing unordered structure (Δ⁹-THC, Δ⁹-THC-OH). Cultured primary hepatocytes from wild-type (WT) mice took up and converted Δ⁹-THC to the above metabolites. Fabp1 gene ablation (LKO) dramatically increased hepatocyte accumulation of Δ⁹-THC and even more so its primary metabolites Δ⁹-THC-OH and Δ⁹-THC-COOH. Concomitantly, rtPCR and Western blotting indicated that LKO significantly increased Δ⁹-THC's ability to regulate downstream nuclear receptor transcription of genes important in both EC ( Napepld > Daglb > Dagla, Naaa, Cnr1) and lipid ( Cpt1A > Fasn, FATP4) metabolism. Taken together, the data indicated that FABP1 may play important roles in Δ⁹-THC uptake and elimination as well as Δ⁹-THC induction of genes regulating hepatic EC levels and downstream targets in lipid metabolism.