Lipid droplets are both highly oxidized and Plin2-covered in hepatocytes of diet-induced obese mice

Abstract

Chronic high-fat diet feeding is associated with obesity and accumulation of fat in the liver, leading to the development of insulin resistance and non-alcoholic fatty liver disease. This condition is characterized by the presence of a high number of intrahepatic lipid droplets (LD), with changes in the perilipin pattern covering them. This work aimed to describe the distribution of perilipin 2 (Plin2), a LD-associated protein involved in neutral lipid storage, and perilipin 5 (Plin5), that is favoring the lipid oxidation in LD, and to evaluate lipid peroxidation through live-cell visualization using the lipophilic fluorescent probe C11-BODIPY581/591 in fresh hepatocytes isolated from high-fat diet (HFD)-fed mice. Male C57BL/6J adult mice were divided into control and HFD groups and fed with a control diet (Control; 10% fat, 20% protein, and 70% carbohydrates) or a HFD (60% fat, 20% protein, and 20% carbohydrates) for 8 weeks. The animals fed the HFD show a significant increase of Plin2 in LD of hepatocytes. LD from HFD-fed mice have a stronger lipid peroxidation level than control hepatocytes. These data provide evidence that obesity status is accompanied by a higher degree of lipid peroxidation in hepatocytes, both in the cytoplasm and in the fats stored inside the LD.
 Novelty bullets
 • Our study shows that lipid droplets from isolated hepatocytes in HFD-fed mice have a stronger lipid peroxidation level than control hepatocytes.
 • C11-BODIPY581/591 is a useful tool to measure the initial level of intracellular lipid peroxidation in single isolated hepatocytes.
 • Perilipins pattern changes with HFD feeding, showing an increase of Plin2 covering lipid droplets.

Effect of Structured Phenolic Lipids with EPA/DHA and Gallic Acid against Metabolic-Associated Fatty Liver Disease (MAFLD) in Mice

“…In the present work, we synthesized bioactive lipids that combine the therapeutic properties of n3-LCPUFA and the antioxidant properties of gallic acid in the same molecule. The study performed on mice fed an HFD to promote the onset of insulin resistance, low-grade steatosis, and obesity showed that the model succeeded in representing the conditions present in MAFLD [48,49]. The results showed that the four-week administration of DRCSO alone could improve insulin levels and the HOMA index with respect to a HFD (Figure 4d,e).…”

Analysis of gene expression changes during lipid droplet formation in HepG2 human liver cancer cells

“…Several studies have reported the association between PLIN2 and liver diseases, and its involvement in fatty liver has been reported in in vivo analysis, since PLIN2 is closely related to fat droplet accumulation in the liver. Nocetti et al (20) reported that hepatocytes from mice with NaFlD induced by a high-fat diet exhibited an increase in Plin2 expression along with highly oxidized fat droplets. Griffin et al (21) also demonstrated that diet-induced hepatic lipidosis was ameliorated in liver-specific Plin2 knockout mice.…”

ITCH E3 ubiquitin ligase downregulation compromises hepatic degradation of branched-chain amino acids

“…Ablation of perilipin 2 (PLIN2), the most abundant lipid droplet-associated protein in steatotic liver, protects mice from diet-induced NAFLD [ 61 ]. Reduced ITCH expression increases Perilipin 2 on lipid droplets, which in turn increases lipid droplets formation in hepatocytes [ 62 , 63 ]. In humans, high levels of Plin2 are present in sarcopenia, hepatic steatosis, atherosclerosis and some types of cancer [ 64 ].…”