Nicotinamide riboside regulates inflammation and mitochondrial markers in AML12 hepatocytes
The effects of nicotinamide riboside on inflammation and mitochondrial biogenesis in a model of hepatocyte steatosis.
The NAD+ precursor nicotinamide riboside (NR) is a type of vitamin B3 found in cow’s milk and yeast-containing food products such as beer. Recent studies suggested that NR prevents hearing loss, high-fat diet-induced obesity, Alzheimer’s disease, and mitochondrial myopathy. The objective of this study was to investigate the effects of NR on inflammation and mitochondrial biogenesis in AML12 mouse hepatocytes. A subset of hepatocytes was treated with palmitic acid (PA; 250 μM) for 48 h to induce hepatocyte steatosis. The hepatocytes were treated with NR (10 μM and 10 mM) for 24 h with and without PA. The cell viability and the levels of sirtuins, inflammatory markers, and mitochondrial markers were analyzed. Cytotoxicity of NR was examined by PrestoBlue assay. Exposure to NR had no effect on cell viability or morphology. Gene expression of sirtuin 1 (Sirt1) and Sirt3 was significantly upregulated by NR in PA-treated hepatocytes. However, Sirt1 activities were increased in hepatocytes treated with low-dose NR. Hepatic pro-inflammatory markers including tumor necrosis factor-alpha and interleukin-6 were decreased in NR-treated cells. NR upregulated anti-inflammatory molecule adiponectin, and tended to down-regulate hepatokine fetuin-A in PA-treated hepatocytes, suggesting its inverse regulation on these cytokines. NR increased levels of mitochondrial markers including peroxisome proliferator-activated receptor γ coactivator-1α, carnitine palmitoyltransferase 1, uncoupling protein 2, transcription factor A, mitochondrial and mitochondrial DNA in PA-treated hepatocytes. These data demonstrated that NR attenuated hepatic inflammation and increased levels of mitochondrial markers in hepatocytes.
In conclusion, our findings demonstrate that NR treatment attenuates hepatic inflammation, and induces mitochondrial biogenesis in mouse hepatocytes. These findings suggest the therapeutic value of NR in hepatic inflammation and impaired mitochondrial biogenesis.