Feeding probiotic Lactobacillus paracasei to Ossabaw pigs on a high-fat diet prevents cholesteryl-ester accumulation and LPS modulation of the Liver X receptor and inflammatory axis in alveolar macrophages

Abstract

Liver X receptors (LXR) play an integral role in cholesterol metabolism and the inflammatory response. High-fat (HF) diets and microbial infection can antagonize the LXR pathway leading to accumulation of cholesteryl-esters (CE) and increased expression of pro-inflammatory mediators in macrophages. The probiotic bacteria Lactobacillus paracasei possesses cholesterol lowering and immune modulating properties. Therefore, the present study sought to model whether daily feeding of L. paracasei to juvenile Ossabaw pigs fed a HF diet could modulate cholesterol metabolism and the LXR/inflammatory axis in lipopolysacharide (LPS)-stimulated alveolar macrophages (AM). The results showed that AM from pigs fed a HF diet had significantly higher concentrations of CE compared to AM from pigs fed a control (C) diet, but not in pigs fed a HF diet with L. paracasei (HFPB). Ex vivo LPS stimulation of AM opposed LXR agonist-mediated transcription of cholesterol metabolism related genes: ABCA1, CH25H and PPARγ in pigs on the C diet, and LXRα, ABCA1, ABCG1, CH25H and PPARγ in pigs on the HF diet. This effect was abrogated for all these genes except LXRα in AM from pigs given L. paracasei. Protein analysis of culture supernatants revealed that AM from HFPB-fed pigs had significantly lower LPS-induced protein expression of IL-1β than AM from HF-fed pigs. Moreover, AM from pigs fed the C diet and given L. paracasei, had significantly higher mRNA levels of IL-8, and IL-6, in response to LPS. These data demonstrated a role for L. paracasei in modulating AM cholesterol metabolism and the response to LPS.

Keywords: ABCA1; ABCG1; AM; ATP-binding cassette, sub-family A, member 1; ATP-binding cassette, sub-family G, member 1; Alveolar macrophages; BAL; Bronchial alveolar lavage; C; CE; CH25H; CPB; Cholesterol; Cholesterol 25-hydroxylase; HDL; HF; HFPB; High density lipoprotein; IL-10; IL-1β; IL-6; IL-8; Interleukin-10; Interleukin-1β; Interleukin-6; Interleukin-8; L. paracasei; LDL; LPS; LXR; LXRs; Lactobacillus paracasei; Liver X receptors; Low density lipoprotein; PPARs; Peroxisome-proliferator-activated receptors; RCT; Reverse cholesterol transport; T09; T0901317; TLR4; TNF-α; Toll-like receptor-4; Tumor necrosis factor-α; alveolar macrophages; cholesteryl-esters; control diet; control plus probiotic diet; high fat diet; high fat plus probiotic diet; lipopolysacharide; liver X receptor.