Vitamin B6 is involved in protein and lipid metabolism. Yet the effect of pyridoxine deficiency has been studied in relation to separate dietary parameters, we aim to investigate the effect of pyridoxine deficiency associated with a high intake of proteins and/or low EFA content in the diet. Vitamin B6 and PLP contents, fatty acid compositions of lipids and phospholipids were determined in liver of groups of rats fed on diets with different amounts of proteins (20-40%) and EFA (2-35%). The results show a complex interaction among tested nutritional factors with a prevailing influence of dietary proteins on lipid content of liver and of low EFA intake and vitamin B6 on PUFA metabolism. The effects of pyridoxine deficiency on lipid metabolism were dependent on the EFA content of the diet.