SMPD3 Inhibition Contributes to Nicotinamide-Ameliorated Hepatic Steatosis in Chronic Alcohol-Fed Mice

Alcohol-associated liver disease (ALD) is characterized by the reduction of hepatic nicotinamide adenine dinucleotide (NAD⁺), which exacerbates hepatic steatosis. The present study was conducted to investigate the protective role of nicotinamide (NAM), a foodborne precursor of NAD⁺ biosynthesis, in ALD. C57BL/6N mice were employed to establish the ALD model and were administered NAM by gavage. Our results showed that NAM supplementation significantly ameliorated alcohol-induced NAD⁺ reduction and lipid accumulation in both mice liver and cultured AML-12 hepatocytes and improved lipid metabolism-associated gene disorders. Alcohol-induced liver injury and oxidative stress were also blocked by NAM administration. Further transcriptomics analysis and validation revealed that alcohol-stimulated sphingomyelin phosphodiesterase 3 (SMPD3) was significantly reversed by NAM, along with the reduction of hepatic ceramide levels. Importantly, SMPD3 was upregulated in the livers of ALD patients. Genetically silencing SMPD3 alleviated alcohol-induced lipid accumulation in hepatocytes. ChIP assay identified SMPD3 as a direct downstream target of hypoxia-inducible factor 1 alpha (HIF-1α). Liver-specific <i>Hif1α</i> knockdown reduced the level of hepatic SMPD3 expression in mice. Activation of HIF-1α abolished the prevention of intrahepatic liver lipid deposition by NAM, while SMPD3 knockdown reversed HIF-1α activation-stimulated lipid accumulation, indicating that a HIF-1α-regulated SMPD3 pathway was involved in the beneficial role of NAM. NAM improved liver oxidative stress, while antioxidant MitoQ administration rescued HIF-1α/SMPD3 activation in ALD mice, implying that the antioxidant effect of NAM contributed to its inhibitory role on the HIF-1α/SMPD3 pathway. In conclusion, NAM ameliorates chronic alcohol intake-induced hepatic steatosis by inhibiting SMPD3. This study provides new insights into the mechanistic understanding of ALD and highlights NAM as a therapeutic choice for ALD treatment.