ISCHOM II: cocoa diet & urinary metabolic profiles. MASSOT-CLADERA

3.6. Metabolic Impact of Cocoa Diet on the Urinary Metabolic Profiles of Rats 

Massot-Cladera, M. *; Mayneris-Perxachs, J.; Swann, J.R.; Costabile, A.; Castell, M.; Pérez-Cano, F.J. 

Background and Objectives 

Cocoa contains fiber and polyphenols that influence the intestinal ecosystem by affecting the growth of certain species of rat’s gut microbiota (Massot-Cladera, M., et al. Arch. Biochem. Biophys. 2012, 527, 105–112; Massot-Cladera, M., et al. Br. J. Nutr. 2014, 112, 1944–1954; Massot-Cladera, M., et al. J. Funct. Foods 2015, 19, 341–352. The aim of the present study was to ascertain whether cocoa also modulates bacterial metabolism and also to establish the particular involvement of cocoa fiber in such effects. 

Methodology

For this purpose, Wistar rats were fed, for three weeks, a standard diet (REF), a diet containing 10% cocoa (providing a final proportion of 0.4% of polyphenols, 0.85% of soluble fiber and 2.55% of insoluble fiber) (C10), a diet with the same soluble and insoluble fiber proportion from cocoa as the C10 diet, but with a very low amount of polyphenols (<0.02%) (CF) or a reference fiber diet (I) containing 0.85% soluble fiber as inulin. After two weeks of intervention, rats were maintained in metabolic cages for 24 h to collect urine output. Urine was centrifuged and frozen at ´80 ˝C until NMR analysis.

Results and Conclusions

There were no differences between the two reference diets (REF and I diets). Animals fed the C10 diet excreted higher levels of epicatechin than those from the I and CF groups. Although both the C10 (OPLSDA model Q2Y = 0.68) and the CF (Q2Y = 0.88) groups excreted more caffeine, theobromine, theophylline, paraxanthine, and malonate than the I group, the C10 diet group excreted higher amounts of these metabolites. Moreover, C10-fed animals excreted higher amounts of 2-oxoglutarate, citrate, choline, allantoin, N-acetylglycoprotein, hippurate, sucrose and glucose than the CF-fed animals. Due to the fact that the C10 diet had a higher impact on the urinary metabolic profile than the CF diet, it is likely that other cocoa compounds, rather than cocoa fiber, are responsible for these effects. 

Acknowledgments

Grants AGL2008-02790 and AGL2011-24279 from the Spanish Ministry of Science and Innovation and the Ministry of Economy and Competitiveness, respectively.


Referencias: ISCHOM II: Barcelona 2015. Nutrients

ISCHOM II: Effects, theobromine, Rats. CAMPOS-BASSACOMA

3.2. Short Term Effects of Cocoa Theobromine on Primary and Secondary Lymphoid Tissues in Rats

Camps-Bossacoma, M. *; Bitlloch-Obiols, M.; Abril-Gil, M.; Franch, A.; Pérez-Cano, F.J.; Castell, M.

Background

Cocoa contains bioactive compounds such as flavonoids, fiber and methylxanthines, mainly theobromine (Katz, D.L., et al. Antioxid, Redox Signal. 2011, 15, 2779–2811; Hurst, W.J., et al. Nature 2002, 418, 289–290). Although many effects of cocoa have been associated with flavonoids (Massot-Cladera, M., et al. Nutrients 2013, 5, 3272–3286), theobromine could also have a role in its effects and this could be reflected in the actions of cocoa on the immune system. Objective: The aim of the present study was to establish the role of cocoa theobromine, in the short term, on the primary (thymus) and secondary (spleen and lymph nodes) lymphoid tissues in rats.

Methodology

Female Lewis rats (three weeks old) were fed either a reference diet, a diet containing 10% cocoa or a reference diet added with the same amount of theobromine provided by cocoa (0.25%). After seven days of free access to water and chow, thymus, spleen and mesenteric lymph nodes (MLN) were removed to isolate lymphocytes to be immune-stained and analyzed by flow cytometry.

Results

In the thymus, lymphocytes were characterized according to the expression of CD4 and CD8 co-receptors. The proportion of CD4+CD8+ (DP) cells was 95% in the reference group and this percentage decreased in animals fed with cocoa and those fed with theobromine. In addition, both nutritional interventions induced an increase in the proportion of CD4+CD8´ with no changes in that of CD4´CD8+ cells. In the spleen, 10% cocoa diet, and also theobromine diet, significantly decreased the TCRγδ+ and NK cell proportions. Although total TCRαβ+ cell percentage was not modified, there was an increase in the T helper lymphocyte proportion and a decrease in T cytotoxic and NKT cell percentage induced by both cocoa and theobromine diets. In MLN, 10% cocoa and 0.25% theobromine diet induced changes in TCRαβ+ subsets: T helper and NKT lymphocyte proportion decreased and, as a consequence, T cytotoxic cells increased. In addition, both dietary interventions increased the proportion of TCRγδ+ CD8αα+.

Conclusion

These results showed that a 10% cocoa diet and a 0.25% theobromine diet produced the same changes in lymphocyte composition in the thymus, spleen and MLN. Therefore, we can conclude that theobromine is the main cacao compound responsible for the changes in lymphoid tissues.

Acknowledgments

AGL2011-24279 from the Spanish Ministry of Economy and Competitiveness.

ISCHOM II: Barcelona 2015. Nutrients