Lipopolysaccharide-binding protein in follicular fluid is associated with the follicular inflammatory status and granulosa cell steroidogenesis in dairy cows

Abstract

Metabolic stress and subsequent hepatic dysfunction in high-producing dairy cows are associated with inflammatory diseases and declining fertility. Lipopolysaccharide (LPS)-binding protein (LBP) is produced by hepatocytes and controls the immune response, suggesting that it is involved in the pathophysiology of inflammation-related attenuation of reproductive functions during metabolic stress. This study investigated the effect of LBP on the inflammatory status, oocyte quality, and steroidogenesis in the follicular microenvironment of dairy cows. Using bovine ovaries obtained from a slaughterhouse, follicular fluid and granulosa cells were collected from large follicles to evaluate the follicular status of metabolism, inflammation, and steroidogenesis. Cumulus-oocyte complexes were aspirated from small follicles and subjected to in vitro embryo production. The results showed that follicular fluid LBP concentrations were significantly higher in cows with fatty livers and hepatitis than in those with healthy livers. Follicular fluid LBP and LPS concentrations were negatively correlated, whereas LPS concentration showed a positive correlation with the concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyric acid in follicular fluid. The blastulation rate of oocytes after in vitro fertilization was impaired in cows in which coexisting large follicles had high NEFA levels. Follicular fluid NEFA concentration was negatively correlated with granulosa cell expression of the estradiol (E₂) synthesis-related gene (CYP19A1). Follicular fluid LBP concentration was positively correlated with follicular fluid E₂ concentration and granulosa cell CYP19A1 expression. In conclusion, follicular fluid LBP may be associated with favorable conditions in the follicular microenvironment, including low LPS levels and high E₂ production by granulosa cells.

Keywords: Dairy cows; Follicles; Lipopolysaccharide binding protein; Oocytes; Steroidogenesis.

Fig. 1.

Follicular fluid lipopolysaccharide-binding protein (LBP), lipopolysaccharide (LPS), non-esterified fatty acids (NEFA), and β-hydroxybutyric acid (BHBA) concentrations of cows with healthy liver, fatty liver, and hepatitis. The numbers in each column indicate the number of follicles used. * P < 0.05 (vs. cows with a healthy liver, one-way analysis of variance followed by Dunnett’s test). Values are presented as the means ± SEM.

Fig. 2.

Relationships between follicular fluid lipopolysaccharide-binding protein (LBP), non-esterified fatty acids (NEFA), and β-hydroxybutyric acid (BHBA) concentrations and lipopolysaccharide (LPS) concentration in cows (Pearson phenotype correlation coefficients). Thirty-one follicles were used for the analysis.

Fig. 3.

Relationships between steroid hormone concentrations and lipopolysaccharide-binding protein (LBP), non-esterified fatty acids (NEFA), and β-hydroxybutyric acid (BHBA) concentrations in follicular fluid of cows (Pearson phenotype correlation coefficients). Thirty-one follicles were used for the analysis.