Figure 1: (a) Representative photomicrograph of the effect of intragastric administration of cocoa butter (0.3 mL) on the lymph volume collected over set periods of 60 min in rat jejunal-originated lymph vessels. The control data is shown before the administration of cocoa butter (b) Summarized data (n=4, the control; 145.0 + 6.5 μL/hr). ** p < 0.01; NS, not significant. (c) Data on the effect of the administration of cocoa butter on the concentration of cells in the lymph are summarized (n=4, the control; 18.3 + 1.1 x 106/mL). ** p < 0.01; NS, not significant.

Figure 2: (a-1) The effects of cocoa butter administration (0.3 mL) on the concentration of IL-1β in the lymph collected over 60 min periods (n=4, the control; 0.0 + 0.0 pg/mL). NS, not significant, * p < 0.05. The control data is shown before the administration of cocoa butter. (b-1) The effects of cocoa butter administration on the concentration of IL-6 in the lymph collected over 60 min periods (n=4, the control; 0.0 + 0.0 pg/mL). * p < 0.05, ** p < 0.01. (c-1) The effects of cocoa butter administration on the concentration of IL-10 in the lymph collected over 60 min (n=4, the control; 0.0 + 0.0 pg/mL). * p < 0.05; NS, not significant. (a-2) The administration of olive oil (0.3 mL) causes no change in IL-1β level in the mesenteric lymph until 240 min after the administration. (b-2) Administration of olive oil releases IL-6 in the mesenteric lymph, being similar to those with cocoa butter intake (the control 0.0 + 0.0 pg/ml, 0 - 60 min 298.3 + 54.3 pg/ml, 60 - 120 min 1670.1 + 653.2 pg/ml, 120 - 180 min 3579.2 + 1293.9 pg/ml, 180 - 240 min 5317.5 + 1114.6 pg/ml; n=4, each value vs the control, p < 0.01 or p < 0.05). (c-2) Administration of olive oil increases slightly IL-10 level in the lymph, being not significantly (n=4, the control; 0.0 + 0.0 pg/mL). NS, not significant.

Figure 3: (a-1) Representative photomicrograph of the effect of pretreatment with clodronate-containing liposomes (20 mg. 24 h) on the cocoa butter-mediated change in the lymph volume over 60 min. The control data is shown before the administration of cocoa butter. (a-2) Summarized data (n=4, the control; 232.5 + 13.8 μL/h). ** p < 0.01. (a-3) The effect of pretreatment with clodronate-containing liposomes on the cocoa butter-mediated change in the concentration of cells in the lymph (n=4, the control; 1.5 + 0.3 x 106/mL). * p < 0.05, ** p < 0.01; NS, not significant.

Figure 4: (a) The effect of pretreatment with clodronate-containing liposomes on the cocoa butter-mediated change in the concentration of IL-1β in the lymph. (n=4, the control; 0.1 + 0.1 pg/mL). NS, not significant. The control data is shown before the administration of cocoa butter. (b) The effect of pretreatment with clodronate-containing liposomes on the cocoa butter-mediated change in the concentration of IL-6 in the lymph. (n=4, the control; 1897.7 + 580.0 pg/mL). * p < 0.05; NS, not significant. (c) The effect of pretreatment with clodronate-containing liposomes on the cocoa butter-mediated response to the concentration of IL-10 in the lymph. (n=4, the control; 50.4 + 18.8 pg/mL). NS, not significant.

Figure 5: (a-1) Representative photomicrographs of the immunoreactivities of the macrophage marker, CD68 in the rat jejunal villi in the absence (left panel) or presence (right panel) of clodronate-containing liposomes (20 mg, 24 h). (a-2) Density measurement of CD68 the immunoreactivity in the absence (left column) or presence (right column) of clodronate (n=10). ** p < 0.01. (b-1) Representative photomicrographs of the immunoreactivity of the macrophage marker, F4/80 in the rat jejunal villi in the absence (left panel) or presence (right panel) of clodronate. (b-2) Density measurement of the immunoreactivity of F4/80 in the absence (left column) or presence (right column) of clodronate(n=10). ** p < 0.01.

Figure 6: (A-a) Two minutes incubation of linoleic acid released IL-1β in the cell medium (n=4, 100% = 43.5 + 13.3 pg/mL, n=4), which significantly reduced after 5 min pretreatment with TLR-2 and -4 inhibitor (5 x 10-6 M, 18.9 + 10.8 %, n=4). ** p < 0.01. The control data is shown before the administration of linoleic acid and TLR 2&4 inhibitor. (A-b) The incubation of linoleic acid released IL-6 in the cell medium (n=4, 100% = 141.9 + 8.9 pg/mL), which significantly reduced after 5 min pretreatment with TLR-2 and -4 inhibitor (5 x 10-6 M, 44.6 + 4.0 %, n=4). ** p < 0.01. (A-c) The incubation of linoleic acid released IL-10 in the cell medium (n=4, 100% = 442.6+ 93.9 pg/mL), which significantly reduced after 5 min pretreatment with TLR-2 and -4 inhibitor (5 x 10-6 M, 45.6 + 7.3 %, n=4). ** p < 0.01. (B-a) Two minutes incubation of linoleic acid released IL-1β in the cell medium (99.8 + 4.0 %, n=4; 100 % = 138.0 + 4.6 pg/mL), which significantly reduced after 5 min pretreatment with MyD 88 (5 x 10-6 M, 18.9 + 10.8 %, n=4). ** p < 0.01. The control data is shown before the administration of linoleic acid and MyD 88 inhibitor. (B-b) The incubation of linoleic acid released IL-6 in the cell medium (100.1 + 33.5 %, n=4; 100 % = 138.0 + 4.6 pg/mL), which was significantly inhibited by MyD 88 (3.2 + 3.2 %, n=4). **p < 0.01. (B-c) Incubation of linoleic acid released IL-10 in the cell medium (100.1 + 33.5 %, n=4; 100 % = 138.0 + 4.6 pg/mL). *p < 0.05. A small amount of IL-10 release was released by phosphate-buffered saline (PBS). (17.2 + 4.5 %, n=4).