After crossing floxed stearoyl-CoA desaturase-1 (Scd1 ^fl/fl) mice with LDL receptor-null (ldlr ^-/-) mice, and then Villin Cre (VilCre) mice, enterocyte Scd1 expression in Scd1 ^fl/fl/ldlr ^-/-/VilCre mice was reduced 70%. On Western diet (WD), Scd1 ^fl/fl/ldlr ^-/- mice gained more weight than Scd1 ^fl/fl/ldlr ^-/-/VilCre mice (P < 0.0023). On WD, jejunum levels of lysophosphatidylcholine (LysoPC) 18:1 and lysophosphatidic acid (LPA) 18:1 were significantly less in Scd1 ^fl/fl/ldlr ^-/-/VilCre compared with Scd1 ^fl/fl/ldlr ^-/- mice (P < 0.0004 and P < 0.026, respectively). On WD, Scd1 ^fl/fl/ldlr ^-/-/VilCre mice compared with Scd1 ^fl/fl/ldlr ^-/- mice had lower protein levels of lipopolysaccharide-binding protein (LBP), cluster of differentiation 14 (CD14), toll-like receptor 4 (TLR4), and myeloid differentiation factor-88 (MyD88) in enterocytes and plasma, and less dyslipidemia and systemic inflammation. Adding a concentrate of tomatoes transgenic for the apoA-I mimetic peptide 6F (Tg6F) to WD resulted in reduced enterocyte protein levels of LBP, CD14, TLR4, and MyD88 in Scd1 ^fl/fl/ldlr ^-/- mice similar to that seen in Scd1 ^fl/fl/ldlr ^-/-/VilCre mice. Adding LysoPC 18:1 to WD did not reverse the effects of enterocyte Scd1 knockdown. Adding LysoPC 18:1 (but not LysoPC 18:0) to chow induced jejunum Scd1 expression and increased dyslipidemia and plasma serum amyloid A and interleukin 6 levels in Scd1 ^fl/fl/ldlr ^-/- mice, but not in Scd1 ^fl/fl/ldlr ^-/-/VilCre mice. We conclude that enterocyte Scd1 is partially responsible for LysoPC 18:1- and WD-induced dyslipidemia and inflammation in ldlr ^-/- mice.