Objective To analyze and compare the effects of multi-oil fat emulsion and medium/long-chain fat emulsion on parenteral nutrition-associated liver disease (PNALD), liver function, and blood lipid indicators in very low birth weight premature infants. Methods A total of 80 very low birth weight premature infants who required long-term parenteral nutrition support (>2 weeks) were retrospectively selected, and they were divided into either a multi-oil fat emulsion group (n=40) or a medium/long-chain fat emulsion group (n=40) according to the type of fat emulsion used. The incidence of PNALD, liver function indicators (including total bilirubin [TBil], direct bilirubin [DBil], total bile acid [TBA], γ-glutamyl transferase [GGT], alkaline phosphatase [ALP], alanine aminotransferase [ALT], aspartate aminotransferase [AST], and albumin [Alb]), and blood lipid indicators (including total cholesterol, triglycerides, low-density lipoprotein cholesterol [LDL-C], and high-density lipoprotein cholesterol [HDL-C]) were compared between the two groups of premature infants. Results The incidence of PNALD in the multi-oil fat emulsion group (0.00%) was lower than that in the medium/long-chain fat emulsion group (15.00%) (P<0.05). The serum levels of TBil, DBil, TBA, GGT, ALP, ALT, and AST in the multi-oil fat emulsion group were all lower than those in the medium/long-chain fat emulsion group, while the serum Alb level was higher than that in the medium/long-chain fat emulsion group (all P<0.05). The serum levels of total cholesterol, triglycerides, and LDL-C in the multi-oil fat emulsion group were all lower than those in the medium/long-chain fat emulsion group, and the serum HDL-C level was higher than that in the medium/long-chain fat emulsion group (all P<0.05). Conclusion Compared with medium/long-chain fat emulsion, the application of multi-oil fat emulsion in parenteral nutrition for very low birth weight premature infants may reduce the incidence of PNALD and improve liver function and blood lipid indicators.