J Rhinol.  2020 May;27(1):8-15. 10.18787/jr.2018.00277.

Polysaccharide from Hizikia Fusiformis Enhances the Immunomodulatory Activity of Macrophages

• Lee J
• Choi MH
• Jang Es
• Shin HJ
• Lee JH

• KMID: 2502788
• DOI: http://doi.org/10.18787/jr.2018.00277

Abstract

Background and Objectives
Hizikia fusiformis is widely used in oriental health food in Japan, China, and Korea, and is known for its anti-oxidation properties. Materials and Method: In this study, we investigated the anti-inflammatory and immune-modulatory effects and mechanisms of Hizikia fusiformis (H. fusiformis) extracts in lipopolysaccharide (LPS)-treated RAW 264.7 cells. RAW 264.7 cells were incubated in the presence of different concentrations of the viscozyme component of H. fusiformis (1, 2, 5, and 10 μg/mL), and changes in expression of pro-inflammatory cytokines (GM-CSF, iNOS, VEGF, and COX-2) were evaluated by real-time PCR and immunoblotting. In addition, the associated signaling pathway including phospho (p)-pNF-κB 65, p-pIkBa, p-p38, and p-p44/42 was also evaluated.
Results
The viscozyme component of H. fusiformis downregulated the expression of GM-CSF, iNOS, VEGF, and COX-2 mRNA. The augmented NO and ROS production was decreased by administration of H. fusiformis. The signal intensity of p-pNF-κB 65, p-pIkBa, p-p38, and p-p44/42 protein activated by LPS was ameliorated by administration of the viscozyme fraction in RAW 264.7 cells.
Conclusion
These results suggest that H. fusiformis has potential as a therapeutic agent for inflammatory diseases.

Keyword

Sargassum; Polysaccharides; NF-kappa B; Inflammation; Immunity

Figure

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  Fig. 1. Effect of H. fusiformis on the viability of RAW 264.7 cells. H. fusiformis extract per se did not affect the viability of airway epithelial cells until the concentration of 100 ug/mL. More than the concentration, the viability was slightly decreased.

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  Fig. 2. Effect of H. fusiformison the phagocytic activity of RAW 264.7 cells. Phagocytic activity of macrophage RAW 264.7cells was significantly increased in a dose-dependent manner (p＜0.05). HF: Hizikia Fusiformis, LPS: lipopolysaccharides.

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  Fig. 3. Effect of H. fusiformis on the NO & ROS production in RAW 264.7 cells. A: RAW 264.7cells were treated with various concentrations of H. fusiformis for 1 h and LPS for 24 h. Nitrite concentration in the supernatant was detected by Griess reagent, and NO production was measured by absorbance at 540 nm. LPS (100 ng/mL) was used as the positive control. Values are expressed as the mean ±SD (n=3, p＜0.05). B: Compared to positive control group, ROS level was decreased in H. fusiformis pre-treated group. NO: nitric oxide, ROS: reactive oxygen species, HF: Hiziki fusiformis, LPS: lipopolysaccharides.

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  Fig. 4. Effect of H. fusiformis on the various mRNA expression. The augmented iNOS mRNA expressions by LPS administration were decreased significantly after H. fusiformis pre-treatment. The elevated levels of GM-CSF and VEGF mRNA by LPS treatment were significantly decreased by H. fusiformis (HF: 10 ug/mL) (p＜0.05). HF: Hiziki fusiformis, LPS: lipopolysaccharides.

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  Fig. 5. Anti-inflammatory effect of H. fusiformis on the MAPK and the NF-κB pathway. The expression of p-p44/42 and p-p38 protein were markedly increased by the treatment of LPS in RAW 264.7 cells, however, pre-treatment of the 10 ug/mL of H. fusiformis decreased their signal intensity. The signal intensity of p-pNF-κB 65 and p-pIκBα was increased by the LPS treatment and their intensity was decreased by the H. Fusiformis administration.

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