Cell surface ATP synthase-released H+ and ATP play key roles in cocoa butter intake-mediated regulation of gut immunity through releases of cytokines in rat

by Selleckchem | Oct 06 2023

Proper food intake is important for maintaining good health in humans. Chocolate is known to exert anti-inflammatory effects; however, the mechanisms remain unclear. In this study, we aimed to investigate the effects of cocoa butter intake on gut immunity in rats and rabbits. Cocoa butter intake increased the lymph flow, cell density, and IL-1β, IL-6 and IL-10 levels in mesenteric lymph. Clodronate, a macrophage depletion compound, significantly enhanced the release of all cytokines. The immunoreactivities of macrophage markers CD68 and F4/80 in the jejunal villi were significantly decreased with clodronate. Piceatannol, a selective cell surface ATP synthase inhibitor significantly reduced the cocoa butter intake-mediated releases of IL-1β, IL-6 and IL-10. The immunoreactivities of cell surface ATP synthase were observed in rat jejunal villi. Shear stress stimulation on the myofibroblast cells isolated from rat jejunum released ATP and carbon dioxide depended with H+ release. In rabbit in vivo experiments, cocoa butter intake increased the concentrations of ATP and H+ in the portal vein. The in vitro experiments with isolated cells of rat jejunal lamina propria the pH of 3.0 and 5.0 in the medium released significantly IL-1β and IL-6. ATP selectively released IL-10. These findings suggest that cocoa butter intake regulates the gut immunity through the release and transport of IL-1β, IL-6, and IL-10 into mesenteric lymph vessels in a negative feedback system. In addition, the H+ and ATP released from cell surface ATP synthase in jejunal villi play key roles in the cocoa butter intake-mediated regulation of gut immunity.

Comments:

The study you described aims to investigate the effects of cocoa butter intake on gut immunity in rats and rabbits. The researchers found that cocoa butter intake led to several effects on the immune system in the gut.

Firstly, cocoa butter intake increased lymph flow, cell density, and the levels of certain cytokines (IL-1β, IL-6, and IL-10) in the mesenteric lymph. The mesenteric lymph is a fluid that carries immune cells and substances through the lymphatic vessels in the mesentery, which is the tissue that supports the intestines.

The researchers also used clodronate, a compound that depletes macrophages (a type of immune cell), to further investigate the effects. They found that clodronate significantly enhanced the release of all cytokines, indicating that macrophages may play a role in regulating the immune response induced by cocoa butter intake. Additionally, the immunoreactivities of macrophage markers CD68 and F4/80 in the jejunal villi (small finger-like projections in the small intestine) were significantly decreased with clodronate treatment, suggesting a decrease in macrophage presence.

Another compound called piceatannol, which selectively inhibits a cell surface ATP synthase enzyme, was used in the study. The researchers found that piceatannol reduced the release of IL-1β, IL-6, and IL-10 induced by cocoa butter intake. This suggests that the release and transport of these cytokines are influenced by the activity of ATP synthase on the cell surface. Immunoreactivities of cell surface ATP synthase were observed in rat jejunal villi, indicating its presence in the gut.

The study also explored the role of shear stress stimulation on myofibroblast cells isolated from the rat jejunum (part of the small intestine). It was found that shear stress stimulation led to the release of ATP, carbon dioxide, and H+ (hydrogen ions). In in vivo experiments with rabbits, cocoa butter intake increased the concentrations of ATP and H+ in the portal vein, which carries blood from the intestines to the liver.

Furthermore, in vitro experiments using isolated cells from the rat jejunal lamina propria (a layer of tissue in the small intestine) showed that low pH levels (pH 3.0 and 5.0) in the medium significantly increased the release of IL-1β and IL-6, while ATP selectively released IL-10. This suggests that pH and ATP release play roles in the regulation of gut immunity mediated by cocoa butter intake.

Overall, the findings suggest that cocoa butter intake may regulate gut immunity through a negative feedback system involving the release and transport of IL-1β, IL-6, and IL-10 into mesenteric lymph vessels. The H+ and ATP released from cell surface ATP synthase in the jejunal villi also appear to play important roles in this regulation.