Anti‐Inflammatory Activity of N‐arachydoyl‐5‐hydroxytryptamine in the Coffee Beans
Authors: Giorno Thais Biondino Sardella , Amorim Jorge Luís , da Silva Moreira Iris Gonçalvez , Rezende Claudia Moraes , Ferdandes Patrícia Dias 
Published: November 2023

Coffee is among the most popular beverages in the world. Some serotonin amides, as N‐arachydoyl‐5‐hydroxytryptamine (C20‐5HT), besides others with long chain acyl units, are present in the surface wax of coffee beans. Few studies have been conducted on the possible biological activity of these amides and an interesting aspect among them is the anti‐inflammatory effect. Thus, the aim of this work was to evaluate the anti‐inflammatory effect of C20‐5HT in the model of carrageenan‐induced inflammation into the subcutaneous air pouch (SAP), production of reactive oxygen species (ROS) by PMA‐stimulated leukocytes and MTT cell viability assay. C20‐5HT present in the Arabica coffee beans (Coffea arabica L., Rubiaceae) was synthesized by the group of Professor Claudia Rezende, Chemistry Institute, UFRJ. Female Webster mice (20–25g; n= 6–8) received oral administration of C20‐5HT (0.1, 1, 3 or 10 mg/kg) and were evaluated in model of leukocyte migration induced by carrageenan into the SAP. Mice received carrageenan injection into the SAP and after 24 h were euthanized and the exsudate from SAP were collected for measurements of cell count, protein, nitric oxide (NO), and cytokines. For the evaluation of ROS, leukocytes from the SAP were collected and incubated with C22‐5HT (0.1, 1 or 3 μM) for 1 h. After, the cells were incubated with 10 nM phorbol myristate acetate and 2 mM 2′‐7′‐dichlorodihidrofluoescein diacetate. The results were expressed as DCF‐DA fluorescence. Cell viability was measured by the MTT assay using RAW 264.7 mouse macrophages. Statistical significance between groups was determined by ANOVA followed by Bonferroni's test (*p<0.05) and the number of the animal research ethical committee is DFBCICB015‐04/16. C20‐5HT showed anti‐inflammatory activity in the two higher doses tested with reduction on cell migration (vehicle= 60.8 ± 8.3 × 106 cells/mL versus 63.2 ± 9.3 × 106 cells/mL, 46.9 ± 7.8 × 106 cells/mL, 30.5 ± 6.6* × 106 cells/mL, and 30.1 ± 7.1* × 106 cells/mL), protein extravasated (vehicle= 264.3 ± 43.4 μg/mL versus 216.5 ± 40.3 μg/mL, 222.6 ± 36.5 μg/mL, 74.1 ± 16.6* μg/mL, and 126.1 ± 25.5* μg/mL) and NO production (vehicle= 141.2 ± 31.8 μM versus 135.3 ± 30.3 μM, 121.2 ± 28.5 μM, 36.4 ± 9.7* μM and 53.1 ± 16.8* μM) with the doses of 0.1, 1, 3, and 10 mg/kg, respectively. It was also observed reduction in levels of cytokines: INF‐γ (vehicle= 122.4 ± 38.6 pg/mL versus 119.5 ± 20.4 pg/mL, 40.5 ± 13.9* pg/mL, 39.7 ± 11.3* pg/mL, 41,5 ± 14.8* pg/mL) and TNF‐α (vehicle= 205.9 ± 31.2 pg/mL versus 196.3 ± 31 pg/mL, 204.3 ± 32.2 pg/mL, 46 ± 11.3* pg/mL, 51.6 ± 13* pg/mL) for 0.1, 1, 3 and 10 mg/kg, respectively. C20‐5HT significantly reduced the production of ROS by PMA‐stimulated leukocytes in all concentrations tested (vehicle= 197,237.4 ± 8,7 DCF‐DA fluorescence versus 103,716.5 ± 10,5* DCF‐DA fluorescence, 69,998.1 ± 7,9* DCF‐DA fluorescence, 75,558.4 ± 7,1* DCF‐DA fluorescence) and did not reduce the cell viability of RAW 264.7 macrophages. Initial results demonstrate that C20‐5HT produces anti‐inflammatory effect, by reducing cell migration, protein extravasation, NO, cytokines production and ROS reduction. The mechanism(s) by which the C20‐5HT produces this effect are still under investigation.Support or Funding InformationTechnical support: Allan MinhoAcknowledgment: IVB (donation of mice)Financial Support: CNPq and FAPERJ