Objective To establish a steatosis model in Mongolian gerbil primary liver cells and to evaluate the effect of Pnpla3 knockdown and Ganoderma lucidum triterpenic acid (GLTA) on lowering lipid content. Methods Primary gerbil liver parenchyma cells were isolated and cultured, and a liver steatosis model was induced by palmitic acid (PA4; 400 μmol / L). Three interfering RNA vectors targeting three Pnpla3 exons were constructed and stably transfected into parenchyma liver cells. The efficiency of Pnpla3 interference was determined. Cells were cultured with high (1 μmol / L), medium (0. 1 μmol / L) and low (0. 01 μmol / L) doses of GLTA and with the positive control drug, methionine (7 μmol / L), and triglyceride (TG) content was determined with and without Pnpla3 knockdown. Results Within 24 hours of induction, PA increased the number of lipid droplets and most cells were of normal shape. siRNA2, designed against the second exon of Pnpla3, had the best knockdown effect and interference efficiency was higher than 70%. Pnpla3 knockdown reduced intracellular TG content. Medium and high dose GLTA had obvious lipid-lowering effects (P< 0. 01), and Pnpla3 knockdown and GLTA had cumulative effects (P< 0. 01). Conclusions A lipid transformation model in primary gerbil hepatocytes was established. GLTA inhibited or reduced the TG content in cells after Pnpla3 knockdown, and the lipid lowering effect of GLTA was associated with the expression of Pnpla3 in two KEGG pathways ( ko00561 and ko00564), which are related to phospholipid metabolism.