Fortifying Butterfat with Soybean Oil Attenuates the Onset of Diet-Induced Non-Alcoholic Steatohepatitis and Glucose Intolerance

Abstract

The addition of plant oils such as soybean oil (S) to a diet rich in saturated fatty acids is discussed as a possible route to prevent or diminish the development of metabolic disease. Here, we assessed whether a butterfat-rich diet fortified with S affects the development of early non-alcoholic steatohepatitis (NASH) and glucose intolerance. Female C57BL/6J mice were fed a standard-control diet (C); a fat-, fructose-, and cholesterol-rich diet (FFC, 25E% butterfat, 50% (wt./wt.) fructose, 0.16% (wt./wt.) cholesterol); or FFC supplemented with S (FFC + S, 21E% butterfat + 4E% S) for 13 weeks. Indicators of liver damage, inflammation, intestinal barrier function, and glucose metabolism were measured. Lipopolysaccharide (LPS)-challenged J774A.1 cells were incubated with linolenic and linoleic acids (ratio 1:7.1, equivalent to S). The development of early NASH and glucose intolerance was significantly attenuated in FFC + S–fed mice compared to FFC-fed mice associated with lower hepatic toll-like receptor-4 mRNA expression, while markers of intestinal barrier function were significantly higher than in C-fed mice. Linolenic and linoleic acid significantly attenuated LPS-induced formation of reactive nitrogen species and interleukin-1 beta mRNA expression in J774A.1 cells. Our results indicate that fortifying butterfat with S may attenuate the development of NASH and glucose intolerance in mice.

Oral Supplementation of Phosphatidylcholine Attenuates the Onset of a Diet-Induced Metabolic Dysfunction–Associated Steatohepatitis in Female C57BL/6J Mice

“…Indeed, we showed before that when using this model of lean MASLD, the development of alterations in glucose metabolism and insulin resistance might take longer than in models of overweight-associated MASLD. 10 , 30 , 31 Further suggesting that in the present study phosphatidylcholine predominantly affected inflammatory alterations, NO levels also were almost at the level of controls in livers of female mice fed the FrFC diet supplemented with phosphatidylcholine. NO has been reported before to be indicative of an induction of inducible nitric oxide synthase in settings of MASLD, 32 and also M1 polarization.…”

“…The sample size was determined based on previous findings. 30 Phosphatidylcholine originating from egg lecithin (purity, 98%; Lipoid, Ludwigshafen, Germany) was used in all experiments. The dose of phosphatidylcholine was based on previous studies by other investigators.…”

“…The dose of phosphatidylcholine was based on previous studies by other investigators. 55 As detailed previously, 30 the intake of the liquid diet was documented daily, and within dietary groups, mice received the amount of diet that was consumed the previous day by the group with the lowest food intake (pair-group). This pair-group had access to diet ad libitum.…”

“…Arginase and MPO activity and NO concentration in liver tissue were assessed as described previously. 30 , 59 In brief, to determine arginase activity, liver tissue was homogenized in 50 mmol/L Tris-HCl, pH 7.5, containing 0.1% Triton X-100, 10 mmol/L MnCl 2 , and protease inhibitor cocktail (Sigma-Aldrich Chemie GmbH), and liver tissue was homogenized in phosphate buffer for measuring MPO activity. NO concentration in liver tissue as well as in cell culture supernatant was measured using a commercially available Griess reagent assay (#G2930; Promega, Mannheim, Germany).…”