This study investigated immunomodulatory and antimetastatic properties of polysaccharides extracted from hallabong (Citrus hybrid) peel, focusing on their effects on macrophage activation. Crude polysaccharides (HPWP) were obtained through hot water extraction and ethanol precipitation from dried hallabong peel powder. Macrophages isolated from mouse peritoneum were used to evaluate cytotoxicity of HPWP against normal cells and Colon 26-M3.1 carcinoma cells. Our findings revealed no cytotoxic effects of HPWP on either normal cells or Colon 26-M3.1 carcinoma cells. Immunoactivity was assessed by measuring cytokine production, specifically IL-6 and IL-12, using ELISA. Results indicated a significant, concentration-dependent increases of cytokine levels. Tumoricidal activity of NK cells was also evaluated, demonstrating enhanced tumor cell killing induced by HPWP, in a concentration-dependent manner. This increased activity was due to HPWP’s potent antimetastatic effects, as observed in a mouse model of Colon 26-M3.1 carcinoma. In comparison with a the negative control (NC, 100%), HPWP at concentrations of 10, 100, and 1000 μg/mouse showed antimetastatic effects of 49.4%, 61.3%, and 74.3%, respectively. These effects were attributed to only NK cell activation. These findings suggest that HPWP has potential as a functional ingredient in food products aimed at enhancing immune response and inhibiting cancer metastasis.

In a previous study, we fractionated crude polysaccharide (AME-CP) with macrophage-stimulating activity from a hot-water extract (AME) of Astragalus membranaceus. AME-CP contained glucose (Glc) as a main component sugar, suggesting that it might be rich in starch-like compounds (SLC). To enhance the immunostimulating activity of AME-CP by pruning SLC rarely known to contribute to activity, hydrolysate (AME-SH) was prepared by digesting with starch-related enzymes, including α-amylase and amyloglucosidase. AME-SH was found to contain substances with molecular weights ranging from 3.9 to 84.4 kDa. These substances were primarily composed of galactose, galacturonic acid, Glc, arabinose, rhamnose, and mannose. AME-SH significantly enhanced the production of macrophage-stimulating factors, including nitric oxide (NO), interleukin (IL)-6, and IL-12, in RAW 264.7 cells compared to AME-CP. Treatment of splenocytes isolated from C3H/HeN mice with AME-SH not only promoted IL-6 secretion, but also induced mitogenic activity. In addition, AME-SH promoted the secretion of hematopoietic growth factors including IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in Peyer's patch (PP) cells and stimulated bone marrow cell proliferation through these PP cells. In conclusion, hydrolysate (AME-SH) digested from AME-CP with starch-related enzymes could be used as a potential immunostimulant.

To utilize pepper (Piper nigrum) as an immunostimulatory agent, we isolated macrophage stimulating polysaccharides from pepper and investigated their macrophage activating activities. Hot-water extracts (HW) of black pepper (BP) and white pepper (WP) were prepared, and their macrophage stimulating activities were evaluated using RAW 264.7 cells. BP-HW significantly promoted the secretion of macrophage stimulating factors such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-12 compared to WP-HW. When BP and WP-HW were fractionated into crude polysaccharides (CP) and low molecules (LM) by ethanol precipitation, BP-CP demonstrated significantly more potent activity. Furthermore, BP-CP not only induced mRNA gene expression of macrophage activation factors, but also promoted nuclear localization of p65 and c-Jun. In addition, component sugar analysis revealed that glucan-type polysaccharides in BP-CP played a crucial role in macrophage activation. Taken together, these findings suggest that black pepper has industrial applicability not only as a spice, but also as an immunostimulatory functional material.

This research investigated the immunoenhancing effect through the intracellular MAPKs and NF-B signaling pathways in macrophages activated by crude polysaccharides (YBP) of barley sprouts. YBP extracted from barley sprouts is composed of xylose (25.8%), arabinose (24.1%), galactose (23.4%), and galacturonic acid (11.7%). YBP did not affect the cytotoxicity and showed superior secretion of nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)- by RAW264.7 cells. Also, YBP dose dependently increased IL-6, TNF-, and inducible nitric oxide synthase (iNOS) mRNA gene expression. In the western blot, YBP strongly induced the phosphorylation of the p38, JNK, ERK, and IB pathways in RAW 264.7 cells. In the anti-pattern recognition receptor (anti-PRRs) assay, the effect of YBP on NO secretion strongly decreased toll-like receptor (TLR) 4 and Dectin1 antibodies, whereas IL-6 and TNF- secretion by YBP mainly decreased SR and CD14. Therefore, we concluded that YBPinduced NO, IL-6, and TNF- were secreted via the MAPKs, while NF-B pathways through TLR4, Dectin1, SR, and CD14 receptors existed in a macrophage surface and were involved in the immunoenhancing effect.

This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

버섯 다당류는 면역 조절 기능, 항암 및 항산화 활성을 비롯하여 항바이러스 활성과 방사선 스트레스의 경감 등 인체에 유익한 다양한 생리활성을 나타낸다. 본 연구에서는 분홍느타리버섯 (Pleurotus djamor var. roseus Corner)으 로부터 뜨거운 물과 에탄올 침전을 이용하여 5.6%의 수율로 갈색을 띠는 경화된 다당류(CPs)를 순차적으로 추출하였다. 이렇게 추출된 CP는 Diethylaminoethyl cellulose (DEAE) 및 sepharose-6B 컬럼 분리를 통해 4개의 분획을 얻었고 각 분획의 총 글루칸 함량은 각각 76.85%, 2.95%, 75.08%, 1.46%로 밝혀졌다. 이중 가장 높은 수율의 분획 (PP)으로부터 300 mg의 백색 분말이 얻어 졌으며, 박층 크로마토그래피(TLC)와 푸리에 변환 적외선 분광법(FTIR) 의 결과로부터 자일로펜토스 유형의 화합물과 함께 다당류 부분의 존재를 확인하였다. PP의 항산화 활성은 1,1- diphenyl-2-picryl-hydrazyl(DPPH) 자유라디칼 소거 분석 및 슈퍼옥사이드 라디칼 소거 분석을 통하여 높은 활성을 나타냄을 확인하였다. PP분획에는 페놀, 단백질 및 단순 탄수화물이 없는 정제된 베타글루칸이 주 구성성분으로, 정 제된 다당류가 천연 항산화제로 사용될 수 있음을 알 수 있었다.

This study observed the anti-inflammatory effect of the polysaccharide derived from the mycelium of Tremella fuciformis in mice with colitis induced with dextran sulfate sodium (DSS). The experimental groups were normal, DSS, DSS-TFL50, DSS-TFH100, and suflasalazine. Body weights, colon lengths, and organ weights were measured, and the plasma level of pro-inflammatory cytokine and mRNA and protein expression in colon tissue were analyzed. Body weight loss, a symptom of DSS-induced colitis, was suppressed by DSS-TF and the speed of weight recovery proceeded rapidly. In addition, DSS-TF showed a significant inhibitory effect on the decrease of colon length typically caused by colon damage. TNF-α, IL-6 and IL-1β cytokine levels in plasma were reduced in DSS-TF and positive control groups. TNF-α, COX-2 and IL-1β mRNA expression in colon tissue were inhibited in DSS-TF and positive control, and it was significantly different from that of the DSS group. The protein expression of inflammation-related genes (IL-6, TNF-α and COX-2) in the colon tissue was significantly increased by DSS compared to that of the normal group, but by DSS-TFL50, DSS-TFH100 and sulfasalarin decreased. In conclusion, the polysaccharide derived from the mycelium of Tremella fuciformis showed the anti-inflammatory effect on DSS-induced colitis in mice.

본 연구에서는, 표고버섯 균사체 발효 생물전환공법으로 생산된 미강생물전환소재(BPP-RB)가 가금티푸스의 주요 원인균인 S. Gallinarum에 감염된 닭 유래 대식세포주 HD- 11에 미치는 효과를 조사하였다. 그 결과, 미강생물전환소 재 추출액은 S. Gallinarum 277에 대한 직접적인 성장억제 효과를 보여주지 않았으며, 총단백질 및 분비단밸질 발현 양상에 어떠한 변화도 유도하지 못하였다. 하지만, 미강생 물전환소재 추출액은 (i) HD-11 대식세포의 탐식 능력 (phagocytic activity)을 활성화하였고, (ii) Th1-type cytokines (tumor necrosis factor-α, interleukin (IL)-1β, iNOS)과 immunosuppressive cytokine IL-10의 발현 증가를 유도하였으며, (iii) Th2-type cytokines (IL-4, IL-6)의 발현은 감 소시키는 것으로 확인되었다. 이러한 결과를 종합하면, 미강생물전환소재는 가금 농장에서 가금티푸스 및 다른 Salmonella종의 감염을 예방하기 위한 사료첨가제로서의 가능성을 가지고 있다고 사료된다.

Sulfated polysaccharides are known to be immune-stimulators in mammals and can be used as food additives to enhance immunity. In this study, the immune-stimulating activity of water-soluble anionic macromolecules F2 fractionation isolated from Codium fragile using ion-exchange chromatography was tested in olive flounder, Paralichythys olivaceus, in vitro and in vivo. The gene expression of interleukin (IL)-1β was adopted to check the immune-affection. As a result, in vitro study revealed that the expression of IL-1β was significantly upregulated in head kidney cells by 1 and 5 μg/ml of polysaccharides 4 h and by 5 μg/ml of polysaccharides at 24 h. In vivo, IL-1β gene expression in head kidney was significantly upregulated by 20 and 100 μg of the polysaccharides at day 1 post-i.p. injection, while downregulated at day 3 but not significant. Meanwhile, in peritoneal cells, it was upregulated by 20 μg of the polysaccharides at day 1 but the upregulation was sustained until day 3 though it was not significant. These results indicate that the sulfated polysaccharides from C. fragile are an immune-stimulator and might be potential feed additives for olive flounder.

The objective of this study was to investigate the effects of combinations of tofu paste and non-starch polysaccharides (NSP) on the oil uptake reduction (OTR) of deep-fat fried cake doughnuts. OTR agents were tofu paste (from grinding tofu with deionized water, followed by passage through a 60 mesh sieve), and five neutral and nine anionic NSPs. A control doughnut (without tofu paste or NSP), tofu doughnut (with tofu paste) and NSP-tofu doughnut (with tofu paste and NSP) were prepared. The moisture and total lipid (TL) content, cross-section image, color characteristic, and specific volume were measured. The tofu and NSP-tofu doughnuts exhibited higher moisture and lower TL content than the control. OTR was 10.8% for the tofu doughnut, and between 13.2% and 41.2% for the NSP-tofu doughnut. The highest OTR (41.2%) was found in the NSP-tofu doughnut with a combination of tofu paste and sodium alginate (NaA). The specific volume of the NSP-tofu doughnuts with combinations of tofu paste with NaA (2.5 mL/g), locust bean gum (2.5 mL/g), and κ-carrageenan (2.4 mL/g) was very close to that of the control (2.6 mL/g). Considering the OTR and specific volume of doughnuts, the combination of tofu paste and NaA would be most effective in reducing the oil uptake of doughnuts during deep-fat frying.

Macrophages play a pivotal role in the innate and adaptive immune systems. This study investigated the immuno-modulatory activities of polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) in macrophages. Polysaccharides from Chrysanthemum zawadskii var. latilobum were extracted by the ethanol precipitation method. RAW 264.7 mouse macrophage cell line was treated with CZPS (4 to 128 μg/mL), and there was no cytotoxicity at a dose below 32 μg/mL. The levels of nitric oxide (NO) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-α and interleukin (IL)-6, IL-1β) production in the CZPS treated group (32 μg/mL) were 6.5±0.12 μM (NO), 1252.8±79.85 (TNF-α), 305.4±29.41 (IL-6), and 683.3±59.71 (IL-1β), respectively, and they were significantly increased when compared to the control group; 2.2±0.03 μM (NO), 452.3±38.34 (TNF-α), 31.7±5.75 (IL-6), and 184.1±11.52 (IL-1β). Additionally, protein expression of inducible nitric oxide synthase (iNOS) and phosphorylation of MAPKs and NF-κB expression were significantly increased upon CZPS treatment. Therefore, these results indicated that polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) may have a potential immunomodulatory activity in macrophages through MAPKs and NF-κB signaling, and this information is useful for the development of immune enhancing adjuvant materials using a natural ingredient.

For the last few decades, especially during summer Korean aquaculture industry has been huge economic loss due to massive blooms caused by Cochlodinium polykrikoides. Moreover, high dispersion speed along the coast and high cell densities (above 10,000 cells ml−1) of C. polykrikoides resuling in red tides for a longer duration (at least for 30 days). C. polykrikoides can be used as a prolific source of extracellular sulfated polysaccharides. Sulfated polysaccharides have showed strong antiviral properties against influenza virus. However, little has been investigated about industrial application of sulfated polysaccharides as a high valuable byproduct from C. polykrikoides. Current commercial microalgal biomass production systems are costly and require advanced instrumental and cultural facilities. From economic point of natural blooms of C. polykrikoides, therefore it is likely to be utilized as a cost-effective way of microalgal biomass production for commercial applications.