Anti-inflammatory potential of carotenoid meso-zeaxanthin and its mode of action

Figures & data

Table 1. Oligonucleotide primer sequences.

Genes	Primer sequence	Product size (bp)
iNOS	Forward 5′-CTTCAACACCAAGGTTGTCTGCAT-3′	231
	Reverse 5′-CAAGACGCGGAAACGAGTACTGTA-3′
COX-2	Forward 5′-GTGGAAAAACCTCGTCCAGA-3′	256
	Reverse 5′-TGATGGTGGCTGTTTTGGTA-3′
TNF-α	Forward 5′-ACTCCCAGAAAAGCAAGCAA-3′	688
	Reverse 5′-T G GAAGACTCCTCCCAGGTA-3′
GAPDH	Forward 5′-CGTCCCGTAGACAAAATGGT-3′	557
	Reverse 5′-CCTTCCACAATGCCAAAGTT-3′

Figure 1. Effect of MZ on carrageenan-induced paw edema formation.

Figure 2. Effect of MZ on dextran-induced paw edema formation.

Figure 3. Effect of MZ on the formalin-induced paw edema formation.

Table 2. Effect of MZ on the production of C-reactive protein and various proinflammatory cytokines by LPS-stimulated macrophages.

	TNF-α (pg/ml)	IL-6 (pg/ml)	IL-1β (pg/ml)	C-reactive protein (pg/ml)
Macrophage alone	34.8 ± 1.1	54.8 ± 2.02	73.7 ± 2.3	248.6 ± 14
Macrophage + LPS	133.2 ± 1.7***	231.1 ± 2***	162.7 ± 1***	1107.9 ± 5***
Macrophage + LPS + vehicle	132.2 ± 0.8^ns	227.1 ± 2.4^ns	165 ± 0.9^ns	1072 ± 6.1^ns
Macrophage + LPS + MZ (5 µg/ml)	60.6 ± 2.2***	87.7 ± 3.2***	95 ± 0.3***	618 ± 3.8***
Macrophage + LPS + MZ (10 µg/ml)	51.2 ± 2.3***	68.9 ± 1.2***	88 ± 0.7***	475 ± 7.4***
Macrophage + LPS + MZ (25 µg/ml)	41.9 ± 1.3***	62.8 ± 1.2***	81 ± 0.8***	403 ± 5.6***
Macrophage + MZ (25 µg/ml)	33.8 ± 0.8***	52.2 ± 0.5***	73 ± 1.2***	243 ± 2.1***

The values were expressed as mean ± standard deviation (SD); n = 6. The mean values were statistically analyzed by one-way analysis of variance (ANOVA) followed by appropriate post hoc test (Dunnett's multiple comparison test). ns, not significant (p > 0.05); ***p < 0.001 significant.

Table 3. Effect of MZ on the production of NO by LPS-stimulated macrophages.

	NO (pg/ml)
Macrophage alone	16.3 ± 0.28
Macrophage + LPS	71.2 ± 2.8***
Macrophage + LPS + vehicle	70.2 ± 1.6^ns
Macrophage + LPS + MZ (5 µg/ml)	48.3 ± 1.9***
Macrophage + LPS + MZ (10 µg/ml)	36 ± 0.7***
Macrophage + LPS + MZ (25 µg/ml)	31.1 ± 1.3***
Macrophage + MZ (25 µg/ml)	16.6 ± 0.2***

The values were expressed as mean ± standard deviation (SD); n = 6. The mean values were statistically analyzed by one-way analysis of variance (ANOVA) followed by appropriate post hoc test (Dunnett's multiple comparison test). ns, not significant (p > 0.05), ***p < 0.001 significant.

Figure 4. Effect of MZ on the expression of TNF-α, iNOS and COX-2 genes in LPS-stimulated macrophages. Lane 1 – Expression of genes in macrophages; Lane 2 – up-regulated expression of genes in LPS-stimulated macrophages; Lane 3 – Down-regulated expression of proinflammatory genes in MZ-treated LPS-stimulated macrophages.