PRODUCTS

Non-alcoholic fatty liver disease (NAFLD) is the most common form of liver disease which is characterized by the abnormal accumulation of fat within the liver. In its most severe form Non-alcoholic steatosis (NASH), the condition can progress to cirrhosis and hepatocellular carcinoma. The most common disease implicated genetic variant is I148M in the gene coding for Patatin-like phospholipase domain containing protein 3 (PNPLA3). The I148M substitution seems to abolish hydrolase activity by reducing substrate access to the enzyme’s active site and so promotes TAG accumulation.

NORMAL

PNPLA3（148M）

Figure. TAG accumulation in PNPLA3 I148M cells.
VLDL, very low density lipoprotein. PMID:27038645. PNPLA3 gene in liver diseases.

BODIPY staining shows that wild-type hepatocytes cultured in low fatty acid media accumulate small amounts of lipids. A few cells are positive for BODIPY staining but most of them show very faint or negative staining. However, after being exposed to high amounts of oleic acid or palmitic acid for 7 days, lipid accumulation increases substantially. In NAFLD model hepatocytes with the PNPLA3 I148M mutation the number of cells with lipids accumulated is higher than in WT even when they are not treated with fatty acids. Both the intensity of staining and distribution of lipids droplets increases when treated with either oleic acid or palmitic acid for 7 days.

Figure 6. Fatty acid accumulation in Def-HEP PNPLA3 cells

BODIPY staining shows a higher accumulation of lipids in Def-HEP I148M even when cultured in low-lipid media.
When cells are treated with 250 μM of Oleic Acid or 250 μM of Palmitic acid for 7 days all cell lines demonstrate an increase in lipid accumulation.