Oleic acid abomasal infusion limits lipolysis and improves insulin sensitivity in adipose tissue from periparturient dairy cows.

Excessive adipose tissue (AT) lipolysis around parturition in dairy cows is associated with impaired AT insulin sensitivity and increased incidence of metabolic diseases. Supplementing cows with oleic acid (OA) reduces circulating biomarkers of lipolysis and improves energy balance. Nevertheless, it is unclear if OA alters lipid trafficking in AT. In the liver and skeletal muscle, OA improves mitochondrial function and promotes lipid droplet formation by activating perilipin 5 (PLIN5) and peroxisome proliferator-activated receptor α (PPARα). However, it is unknown if this mechanism occurs in AT. The objective of this study was to determine the effect of OA on AT lipolysis, systemic and AT insulin sensitivity, and AT mitochondrial function in periparturient dairy cows. Twelve rumen-cannulated Holstein cows were infused abomasally following parturition with ethanol (CON) or OA (60 g/d) for 14 d. Subcutaneous AT samples were collected at 11 ± 3.6 d before calving (-12 d), and 6 ± 1.0 d (7 d) and 13 ± 1.4 d (14 d) after parturition. An intravenous glucose tolerance test was performed on d 14. Adipocyte morphometry was performed on hematoxylin and eosin-stained AT sections. The antilipolytic effect of insulin (1 µg/L) was evaluated using an ex vivo explant culture following lipolysis stimulation. PLIN5 and PPARα transcription and translation were determined by real-time quantitative PCR and capillary electrophoresis, respectively. RNA sequencing was used to evaluate the transcriptomic profile of mitochondrial gene networks. In CON cows, postpartum lipolysis increased the percentage of smaller (<3,000 µm2) adipocytes at 14 d compared with -12 d. However, OA limited adipocyte size reduction at 14 d. Likewise, OA decreased lipolysis plasma markers nonesterified free fatty acids and ß-hydroxybutyrate at 5 and 7 d. Over the 14-d period, compared with CON, OA increased the concentration of plasma insulin and decreased plasma glucose. During the glucose tolerance test, OA decreased circulating glucose concentration (at 10, 20, 30, 40 min) and the glucose clearance rate. Moreover, OA increased insulin at 10 and 20 min and tended to increase it at 30 min. Following lipolysis stimulation, OA improved the antilipolytic effect of insulin in the AT at 14 d. PLIN5 and PPARA gene expression decreased postpartum regardless of treatment. However, OA increased PLIN5 protein expression at 14 d and increased PPARA at 7 and 14 d. Immunohistochemical analysis of AT and RNA sequencing data showed that OA increased the number of mitochondria and improved mitochondrial function. However, OA had no effect on production and digestibility. Our results demonstrate that OA limits AT lipolysis, improves systemic and AT insulin sensitivity, and is associated with markers of mitochondrial function supporting a shift to lipogenesis in AT of periparturient dairy cows.

Abomasal infusion of different exogenous emulsifiers alters fatty acid digestibility and milk fat yield of lactating dairy cows.

We evaluated the effects of abomasal infusion of emulsifiers on fatty acid (FA) digestibility and milk production of lactating dairy cows. All emulsifiers examined were polysorbates, nonionic surfactants, consisting of a polyethoxylated sorbitan esterified with FA. The polysorbates tested in this study consisted of the same polyethoxylated sorbitan base but differed by the FA esterified to it. Eight rumen-cannulated multiparous cows (89 ± 13 d in milk) were assigned to a treatment sequence in 4 × 4 Latin squares with 18-d periods consisting of 7 d of washout and 11 d of infusion. Treatments were abomasal infusions of water only (CON) or 30 g/d of different emulsifiers as follows: polysorbate-C16:0 (T40), polysorbate-C18:0+C16:0 (T60), and polysorbate-C18:1 (T80). Emulsifiers were dissolved in water and delivered at 6-h intervals (total daily infusion was divided into 4 equal infusions per day). Cows were fed the same diet that contained (% diet dry matter) 32.1% neutral detergent fiber, 15.7% crude protein, 25.8% starch, and 3.32% FA (including 1.92% FA from a saturated FA supplement containing 34.2% C16:0 and 47.7% C18:0). The T80 treatment increased total FA digestibility compared with CON (5.40 percentage units) and T60 (3.90 percentage units) and tended to increase it compared with T40. Also, T40 tended to increase and T80 increased (4.80 percentage units) 16-carbon FA digestibility compared with CON. The T80 treatment increased 18-carbon FA digestibility compared with the other treatments. The T40 treatment tended to increase and T80 increased total FA absorption compared with CON (53 g/d) and T60 (52 g/d). Both T40 and T80 increased the absorption of 16-carbon FA compared with CON and T60. The T60 treatment did not differ from CON for any digestibility variable. Both T40 and T80 increased the yields of milk fat, 3.5% fat-corrected milk, and de novo, mixed, and preformed milk FA compared with CON. In conclusion, not all emulsifiers increased FA digestibility. Compared with CON, T80 increased the digestibility and absorption of total, 16-, and 18-carbon FA. The T40 treatment tended to increase and T80 increased total FA absorption and the yields of milk fat and 3.5% FCM compared with CON. Milk fat yield was increased by increases in de novo, mixed, and preformed milk FA. In our short-term infusion study, results suggest that the predominant FA present in the polysorbate affects its ability to improve FA digestibility. Overall, FA digestibility and absorption were improved the most when cows received the T80 treatment.

Lipopolysaccharide induces lipolysis and insulin resistance in adipose tissue from dairy cows.

Intense and protracted adipose tissue (AT) fat mobilization increases the risk of metabolic and inflammatory periparturient diseases in dairy cows. This vulnerability increases when cows have endotoxemia-common during periparturient diseases such as mastitis, metritis, and pneumonia-but the mechanisms are unknown. Fat mobilization intensity is determined by the balance between lipolysis and lipogenesis. Around parturition, the rate of lipolysis surpasses that of lipogenesis, leading to enhanced free fatty acid release into the circulation. We hypothesized that exposure to endotoxin (ET) increases AT lipolysis by activation of classic and inflammatory lipolytic pathways and reduction of insulin sensitivity. In experiment 1, subcutaneous AT (SCAT) explants were collected from periparturient (n = 12) Holstein cows at 11 ± 3.6 d (mean ± SE) before calving, and 6 ± 1 d and 13 ± 1.4 d after parturition. Explants were treated with the endotoxin lipopolysaccharide (LPS; 20 µg/mL; basal = 0 µg/mL) for 3 h. The effect of LPS on lipolysis was assessed in the presence of the ß-adrenergic agonist and promoter of lipolysis isoproterenol (ISO; 1 µM; LPS+ISO). In experiment 2, SCAT explants were harvested from 24 nonlactating, nongestating multiparous Holstein dairy cows and exposed to the same treatments as in experiment 1 for 3 and 7 h. The effect of LPS on the antilipolytic responses induced by insulin (INS = 1 µL/L, LPS+INS) was established during ISO stimulation [ISO+INS, LPS+ISO+INS]. The characterization of lipolysis included the quantification of glycerol release and the assessment of markers of lipase activity [adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), and phosphorylated HSL Ser563 (pHSL)], and insulin pathway activation (AKT, pAKT) using capillary electrophoresis. Inflammatory gene networks were evaluated by real-time quantitative PCR. In periparturient cows, LPS increased AT lipolysis by 67 ± 12% at 3 h across all time points compared with basal. In nonlactating cows, LPS was an effective lipolytic agent at 3 h and 7 h, increasing glycerol release by 115 ± 18% and 68.7 ± 16%, respectively, relative to basal. In experiment 2, LPS enhanced ATGL activity with minimal HSL activation at 3 h. In contrast, at 7 h, LPS increased HSL phosphorylation (i.e., HSL activity) by 123 ± 11%. The LPS-induced HSL lipolytic activity at 7 h coincided with the activation of the MEK/ERK inflammatory pathway. In experiment 2, INS reduced the lipolytic effect of ISO (ISO+INS: -63 ± 18%) and LPS (LPS+INS: -45.2 ± 18%) at 3 h. However, the antilipolytic effect of INS was lost in the presence of LPS at 7 h (LPS+INS: -16.3 ± 16%) and LPS+ISO+INS at 3 and 7 h (-3.84 ± 23.6% and -21.2 ± 14.6%). Accordingly, LPS reduced pAKT:AKT (0.11 ± 0.07) compared with basal (0.18 ± 0.05) at 7 h. Our results indicated that exposure to LPS activated the classic and inflammatory lipolytic pathways and reduced insulin sensitivity in SCAT. These data provide evidence that during endotoxemia, dairy cows may be more susceptible to lipolysis dysregulation and loss of adipocyte sensitivity to the antilipolytic action of insulin.

Replacing stearic acid with oleic acid in supplemental fat blends improves fatty acid digestibility of lactating dairy cows.

The objective of our study was to determine the effects of altering the ratio of stearic (C18:0; SA) and oleic (cis-9 C18:1; OA) acids in supplemental fatty acid (FA) blends on FA digestibility and milk yield of dairy cows. Eight multiparous Holstein cows (mean ± SD; 157 ± 11.8 d in milk) were randomly assigned to treatment sequence in a replicated 4 × 4 Latin square design with 14-d periods. Digestibility and production data were collected during the last 4 d of each period. The treatments were an unsupplemented control diet (CON), and 3 diets incorporating FA supplement blends at 1.4% of diet dry matter (DM) containing (as a % of total FA) 50% SA and 10% OA, 40% SA and 20% OA, or 30% SA and 30% OA. The FA blends were balanced to contain 33% palmitic, 5% linoleic, and <0.5% linolenic acids. The FA supplements replaced soyhulls in the CON diet. Preplanned contrasts were as follows: (1) overall effect of FA treatments [CON vs. the average of the FA-supplemented diets; (50:10 + 40:20 + 30:30)/3], (2) the linear effect of OA inclusion in the supplemental FA blend, and (3) the quadratic effect of OA inclusion in the supplemental FA blend. There was no effect of treatment on DM intake, but the replacement of soyhulls in the FA treatments decreased neutral detergent fiber intake. Overall, compared with CON, FA treatments increased DM and neutral detergent fiber digestibility, and increasing OA within FA treatments quadratically increased digestibility of DM and neutral detergent fiber. Overall, FA treatments increased the intake of total, 16-carbon, and 18-carbon FA, decreased the digestibility of total and 18-carbon FA, but increased absorption of total, 16-carbon, and 18-carbon FA. Within FA treatments, increasing OA linearly increased the digestibility of total, 16-carbon, and 18-carbon FA, as well as the absorption of total, 16-carbon, and 18-carbon FA. Overall, FA treatments increased the yields of milk, energy-corrected milk, and milk fat, and tended to increase milk protein yield. Compared with CON, FA treatments had no effect on the yield of de novo milk FA and increased the yields of mixed and preformed milk FA. Within FA treatments, increasing OA did not affect the yields of milk or milk components, linearly decreased the yield of de novo FA, and quadratically affected the yield of mixed and preformed milk FA. Overall, FA treatments increased plasma nonesterified fatty acids but did not affect ß-hydroxybutyrate or insulin. Within FA treatments, increasing OA quadratically affected plasma nonesterified fatty acids, and tended to linearly increase ß-hydroxybutyrate and quadratically affect insulin. In conclusion, supplemental FA blends containing different ratios of SA and OA did not affect DM intake but increased the yields of milk and milk components. Supplemental FA blends also increased digestibility of DM and neutral detergent fiber and decreased digestibility of total and 18-carbon FA compared with CON. Although increasing OA within FA supplements did not alter milk production, increasing OA within FA supplements increased total, 16-carbon, and 18-carbon FA digestibility and FA absorption. Further research is required to determine longer term effects of SA and OA on nutrient digestion and partitioning and opportunities for maintaining or improving FA digestibility with increasing SA intake and availability in the small intestine.

Transcriptomic profiling of adipose tissue inflammation, remodeling, and lipid metabolism in periparturient dairy cows (Bos taurus).

BACKGROUND: Periparturient cows release fatty acid reserves from adipose tissue (AT) through lipolysis in response to the negative energy balance induced by physiological changes related to parturition and the onset of lactation. However, lipolysis causes inflammation and structural remodeling in AT that in excess predisposes cows to disease. The objective of this study was to determine the effects of the periparturient period on the transcriptomic profile of AT using NGS RNAseq. RESULTS: Subcutaneous AT samples were collected from Holstein cows (n = 12) at 11 ± 3.6 d before calving date (PreP) and at 6 ± 1d (PP1) and 13 ± 1.4d (PP2) after parturition. Differential expression analyses showed 1946 and 1524 DEG at PP1 and PP2, respectively, compared to PreP. Functional Enrichment Analysis revealed functions grouped in categories such as lipid metabolism, molecular transport, energy production, inflammation, and free radical scavenging to be affected by parturition and the onset of lactation (FDR < 0.05). Inflammation related genes such as TLR4 and IL6 were categorized as upstream lipolysis triggers. In contrast, FASN, ELOVL6, ACLS1, and THRSP were identified as upstream inhibitors of lipid synthesis. Complement (C3), CXCL2, and HMOX1 were defined as links between inflammatory pathways and those involved in the generation of reactive oxygen species. CONCLUSIONS: Results offer a comprehensive characterization of gene expression dynamics in periparturient AT, identify upstream regulators of AT function, and demonstrate complex interactions between lipid mobilization, inflammation, extracellular matrix remodeling, and redox signaling in the adipose organ.