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ARS Home » Midwest Area » Lexington, Kentucky » Forage-animal Production Research » Research » Publications at this Location » Publication #413959
Research Project: Increasing Sustainability of Forage Production in Mid-South Agroecosystems

Location: Forage-animal Production Research

Title: The effects of ß-caryophyllene on butyrate utilization and metabolism in Caco 2 cells

Author
item SCROGGINS, HANNAH - University Of Kentucky
item KENT-DENNIS, CORAL - Orise Fellow
item MAY, JOHN - University Of Kentucky
item Klotz, James
item HARMON, DAVID - University Of Kentucky

Submitted to: American Society of Animal Science Southern Section Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/11/2024
Publication Date: 9/18/2024
Citation: Scroggins, H., Kent-Dennis, C., May, J., Klotz, J.L., Harmon, D.L. 2024. The effects of ß-caryophyllene on butyrate utilization and metabolism in Caco 2 cells. In: Journal of Animal Science. American Society of Animal Science Southern Section Meeting, January 27-30, 2024, Louisville, Kentucky. p. 265-266.

Interpretive Summary:

Technical Abstract: Butyrate is important for regulating energy status and cellular metabolism. Beta-caryophyllene (BCP) is a plant compound which may exert potentially beneficial effects on intestinal epithelial cells, such as barrier integrity and modulation of nutrient utilization. The goal of this preliminary study was to investigate the effects of BCP on butyrate utilization and metabolism in intestinal epithelial cells. Caco 2 cells were used as a model for the intestinal barrier. Cells were seeded in multi-well plates with hanging inserts (n=4) and grown for 18d. On d 19, 1 of 4 treatments was added to triplicate wells per plate. Treatments included vehicle control (VC), 40 µM BCP (BCP), 2 mM butyrate (BUT), and butyrate plus BCP (BB). Treatments were added to the apical side and incubated for 24 and 48 h. Transepithelial electrical resistance (TEER; ohms/cm2) readings and supernatant samples from both the apical and basolateral sides of the cell layer were taken at each time point, and butyrate and BHB levels measured. Butyrate utilization was calculated as absolute nmol (nanomoles) lost. Statistical analyses were performed using lm and emmeans packages of R, with treatment, time and side (of cell layer) as fixed effects. All wells started with the same statistical TEER values on hour 0. After 24 and 48h BCP showed no changes in TEER reading (P>0.05). Compared to VC, the mean TEER for BUT and BB was greater at 24h (579 vs 878 and 1008 ± 35, respectively; P<0.001). After 48h, all groups returned to baseline, except BB which had a greater TEER (883 ± 35; P<0.001) compared to other treatments. After 24 and 48h both sides of the cell layer, BCP showed no changes (P>0.05). Overall, absolute BHB levels increased from 24 to 48h in all treatments (P<0.05). Production of BHB in each of the VC and BCP groups was less than 15 nmol at each time point. For BB and BUT groups, more nmol BHB was detected on the apical (14.0 and 10.4 ± 1.3) vs basolateral side (31.9 and 22.2 ± 1.3, respectively; P<0.001) after 24h. After 48h, BHB was also higher on the apical side (30.2 and 24.0 ± 1.3 nmol) vs basolateral side (58.2 and 45.2 ± 1.3 nmol for BB and BUT respectively; P<0.001). When looking at butyrate utilization, the BB treatment had greater disappearance than BUT in 24h (254 vs 169 ± 12.6 nmol; P=0.0005) and 48h (707 vs 573 ± 12.6 nmol; P<0.0001). BCP may promote the utilization and metabolism of butyrate by intestinal epithelial cells. More research should be done in order to understand the mechanism of how BCP can play a role in cellular butyrate utilization.