The leaves and stalks of sweet potato have attracted considerable interest as a health food due to numerous studies reporting the presence of functional compounds and various physiological activities. This study analyzed the functional components in the aerial parts of six domestically developed sweet potato cultivars and compared their antioxidant activities. The total polyphenol content, total flavonoid content, and total phenolic acid content ranged from 76.9 to 148.6 mg GAE/g, 3.98 to 11.90 mg CE/g, and 44.19 to 93.56 mg/100 g, respectively. Among the cultivars examined, 'Gogeonmi' and 'Tongchaeru' exhibited high levels of these compounds, and their DPPH and ABTS radical scavenging activities were superior to those of the other cultivars. The GABA content ranged from 0.59 to 2.55 mg/g, with 'Tongchaeru' and 'Jinhongmi' showing the highest levels. Lutein content ranged from 0.10 to 0.24 mg/g, with 'Tongchaeru' reaching its maximum concentration 90 days post-cultivation. Extracts from 'Tongchaeru' significantly inhibited the secretion of interleukin-6 (IL-6) inflammatory cytokines, with the water extract demonstrating a stronger effect than the pretanol extract. These findings suggest that the aerial parts of sweet potato could serve as excellent functional vegetables and bioactive ingredients for health food applications.

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.

The risk of inflammatory conditions caused by obesity is associated with an increased predisposition for additional pathological conditions, including cardiovascular risk factors. Adipose tissue stores energy and contributes to endocrine and immune functions that regulate homeostasis throughout the body. The effects of honokiol on vascular homeostasis in adipose tissue in high-fat diet (HFD)-induced obese mice are unclear. This study examined the protective effect of honokiol, an extract of traditional alkaloid herbs, on vascular endothelial cells in epididymal adipose tissue (EAT) and its regulatory effect on other metabolic parameters, such as the lipid droplet diameter, macrophage infiltration, and inflammation in HFDinduced obese mice. A HFD increased the density of platelet endothelial cell adhesion molecule-1 (PECAM-1)-1-positive vascular endothelial cells in EAT, which was decreased significantly by the honokiol treatment. Honokiol ameliorated the HFD-induced increase in lipid droplet diameter and increased macrophage infiltration in adipose tissue. Honokiol ameliorated the up-regulation of pro-inflammatory molecules and F4/80-positive macrophage infiltration in the adipose tissue of HFD-induced obese mice. Obese mice administered honokiol exhibited reduced mRNA expression of M1 macrophage (F4/80, TNF-, mIL-1, CD11c, and CCL2) and M2 macrophage (Arginase-1, FIZZ1, CD206, and TGF-1) markers in EAT. The vascular permeability was detected by Evans blue dye leakage in EAT of obese mice and treated mice with honokiol. These data suggest that honokiol regulates the angiogenic effects in adipose tissue and inflammation in HFDinduced obese mice.

This study investigated major constituents and anti-inflammatory effects of an ethanol extract of Platycodon grandiflorum leaves. Through HPLC analysis, chlorogenic acid and luteolin-7-O-glucoside were identified as predominant constituents in the ethanol extract. Their anti-inflammatory effects were evaluated using murine macrophage (RAW 264.7 cells) and human lung carcinoma cells (NCI-H292 & A549). The ethanol extract significantly (p<0.01) inhibited the production of nitrite, interleukin-6 (IL-6), and prostaglandin E2 (PGE2) induced by lipopolysaccharide (LPS) in RAW 264.7 cells. Furthermore, the ethanol extract suppressed the expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) proteins in RAW 264.7 cells stimulated with LPS. In NCI-H292 and A549 cells, treatment with the ethanol extract significantly (p<0.05) decreased levels of pro-inflammatory cytokines IL-6 and IL-8 induced by IL-1β. The phosphorylation of ERK rather than JNK in the mitogen-activated protein kinase signaling pathway was observed to be a more important mediator in the down-regulation of pro-inflammatory cytokines in NCI-H292 cells. These findings suggest that the ethanol extract of Platycodon grandiflorum leaves containing luteolin-7-O-glucoside exhibits promising anti-inflammatory properties.

본 연구는 국내산 가시오가피의 건강기능식품 소재로서 의 가능성을 확인하기 위해 산지별 채취한 가시오가피의 유효물질 함량 및 면역 증강 효과을 평가하였다. 태백, 철 원, 삼척, 강원도 농업기술원에서 수확한 가시오가피의 지 표성분인 eleutheroside B 및 eleutheroside E의 분석을 수 행하였으며, 면역 증강에 대한 효과를 관찰하기 위하여 MTT 세포독성 평가, NO 생성량과 cytokine 생성량을 측 정하였다. 지표성분 eleutheroside B의 함량은 채취 지역별 로 70% 에탄올 추출물에서는 2.96±0.11-6.24±0.05 mg/g로 태백에서 가장 높은 함량을 나타냈으며, 열수 추출물에서 는 1.11±0.05-2.11±0.03 mg/g로 태백에서 가장 높은 함량 을 나타냈다. Eleutheroside E 함량은 채취 지역별로 70% 에탄올 추출물에서는 4.93±0.20-10.79±0.03 mg/g을 나타냈으 며 철원에서 가장 높은 함량을 나타냈고, 열수 추출물에서 는 1.75±0.14-3.64±0.05 mg/g로 철원과 농업기술원에서 가장 높은 함량을 나타냈다. 또한, eleutheroside B 및 E 함량은 열수 추출물보다 70% 에탄올 추출물에서 더 높은 함량을 나타냈다. 채취 지역별 가시오가피의 70% 에탄올 추출물은 50-200 μg/mL 농도에서, 열수 추출물은 100-500 μg/mL 농도 에서 RAW 264.7 대식세포에 대한 세포독성을 나타내지 않 았으며, 대식세포의 활성화로 방출되는 NO 성성량을 측정 한 결과, 가시오가피 줄기 추출물에서 NO 생성량이 증가하 는 것을 확인하였으며, TNF-α, IL-6, IL-1β을 포함하는 cytokine의 방출을 측정한 결과 유의적인 증가를 나타냈다. 따라서 가시오가피 줄기는 면역 관련 질환의 개선을 위한 건강기능식품 소재로 활용 가능할 것으로 사료된다.

To increase industrial applicability of Astragalus membranaceus (AM) as immunostimulating materials, hot-water extract (AME) was prepared from AM and fermented with Kimchi-lactic acid bacteria (Lactobacillus sakei & Leuconostoc mesenteroides) to prepare fermented AM-postbiotics (FAME). Although FAME prepared from AM-postbiotics did not show a significant enhancement in macrophage stimulating activity compared to non-fermented AME, crude polysaccharide (FAME-CP) fractionated by EtOH precipitation from FAME showed significantly higher macrophage stimulating activity than AME-CP. Compared to AME-CP, FAME-CP showed dramatic changes in component sugar and molecular weight distribution. FAME-CP was a polysaccharide with a major molecular weight distribution of 113.4 kDa containing Man (44.2%), Glc (19.3%), Gal (10.2%), GalA (10.2%), and Ara (7.4%) as sugar components. FAME-CP with enhanced macrophage stimulatory activity not only increased expression levels of mRNA genes encoding macrophage-activated factors (iNOS, TNF-α, MCP-1, IL-6, and COX-2), but also led the nuclear translocation of activated p65 and c-Jun. In conclusion, crude polysaccharide from AM-postbiotics fermented with lactic acid bacteria could increase industrial applicability as a functional material with enhanced immunostimulating activity than AME-CP.

We investigated the anti-inflammatory effects of shiitake mushroom and kelp (SMK) mixture extracts in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 cells. Treatment of RAW 264.7 cells with LPS significantly increased NO (nitric oxide) production, pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and IL-1β), and inflammation-related genes (COX-2 and inducible nitric oxide synthase (iNOS)). In cytotoxicity testing using RAW 264.7 cells, SMK mixture extracts in the range of 1-16 μg/mL did not inhibit cell proliferation. However, SMK mixture extracts significantly inhibited NO production in a dose-dependent manner (p<0.05). SMK treatment significantly decreased TNF-α, IL-6, IFN-γ, and IL-1β levels compared to the LPS group, and similarly, pro-inflammatory cytokine mRNA levels also decreased. SMK mixture extracts reduced the mRNA expression of COX-2 and iNOS in RAW 264.7 cells compared to LPS (p<0.05). The above results show that SMK mixture extracts suppressed the inflammatory response induced by LPS. In particular, the extracts were shown to regulate the inflammatory response by suppressing the expression of inflammatory cytokines and inflammation-related enzymes.

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.

To enhance the bioavailability and bioactivities of mixed herbal medicines (RW), they were fermented with lactic-acid bacteria isolated from kimchi into postbiotics (FRW). Then, from the results of the 16s rRNA sequencing analysis, lactic acid bacteria isolated from kimchi were identified to be of two species, namely Lactobacillus sakei and Leuconostoc mesenteroides. The FRW prepared from the RW were extracted using hot water (HW) and 70% EtOH (EtOH) for comparison of their macrophage-stimulating activities. Based on a comparison of the activities of the FRW extracts, nitric oxide (NO) production of HW was significantly higher than that in EtOH. An analysis of the chemical properties of the extracts showed that HW had higher contents of neutral sugar and uronic acid than EtOH as well as contained a large amount of glucose. In addition, crude polysaccharide (CP) was prepared to enhance the macrophage-stimulating activity. The FRW-CP not only secreted immunostimulatory mediators but also increased the expression of immunostimulatory genes (iNOS, TNF-α, MCP-1, and IL-6). The fractionated FRW-CP contained about 90% neutral sugars, and these sugars were mainly composed of glucose, galacturonic acid, and arabinose. Thus, FRW prepared by fermentation of RW with kimchi lactic acid bacteria were found to be immunostimulatory modulators.

Asthma is a chronic inflammatory disease characterized by recurring symptoms, airflow obstruction, and bronchial hyper-responsiveness. The onset of asthma for most patients begins early in life, and current asthma treatment with anti-inflammatory agents can have adverse effects, eventually leading to impaired quality of life. In the pathogenesis of asthma, macrophages and basophils play a vital role during progression. Macrophages not only induce inflammation by secreting inflammatory cytokines but also promote DNA damage and mucus production through nitric oxide (NO) production. Basophils enhance eosinophil recruitment and aggravate asthma through the FcεRIα receptor with high affinity for histamine and IgE. Therefore, in this study, we investigated whether the activation of macrophages and basophils is suppressed by the individual extracts of 28 natural products. RAW 264.7 cells (mouse macrophages) were treated with the natural products in LPS, and 4 natural product extracts resulted in decreased NO production. In β-hexosaminidase assay using RBL-2H3 cells (rat basophils), 19 natural product extracts decreased β-hexosaminidase production. In NO production and β-hexosaminidase assay using macrophages and basophils, 3 natural product extracts (Plantago asiatica, Centella asiatica, and Perilla frutescens var. japonica) significantly inhibited NO production and β-hexosaminidase release. Overall, we examined the inhibitory effects of 28 natural product extracts on macrophage and basophil activity, and the findings demonstrated the potential of natural product extracts for treating asthma and macrophage- and basophil-related diseases.

The purpose of this study was to analyze the anti-inflammatory effect of Chung-Dae Indigo Pulverata Levis, indigo naturalis) produced during indigo dyeing. As a result of in vitro cytotoxicity experiments using RAW 264.7 cell, Chung-Dae extract did not inhibit cell proliferation in Raw 264.7 cells in the range of 1~32 μg/mL. NO production was significantly reduced when Chung-Dae extracts were treated at concentrations of 2, 8, and 32 μg/mL (p<0.05). The pro-inflammatory cytokines TNF-α, IL-6, IL-1β and IFN-γ significantly decreased when the Chung-Dae extract was treated at concentrations of 2, 8, and 32 μg/mL compared to the LPS group, and similarly, the TNFα and IL-6 mRNA levels also decreased. Additionally, the mRNA level of COX-2 was also suppressed. At the protein expression level, the expression of TNF-α, IL-6, iNOS and COX-2 were observed with LPS and Chung-Dae extract significantly decreased compared to the group treated with only LPS (p<0.05). From the above results, it shows that Chung-Dae extract, a plant-derived compound, inhibits the inflammatory response induced by LPS in RAW 264.7 cells. and in particular, regulates the inflammatory response by inhibiting the expression of pro-inflammatory cytokines and inflammation-related enzymes.

Maca roots (Lepidium meyenii) are an important medicinal herb that have long been used by the Andes-indigenous peoples and South Americans. In Korea, recently, it has attracted attention as a health food material because of nutritional values and physiological activities. The purpose of this study was to investigate the industrial applicability of maca (red and golden varieties; R&G) as immunostimulating materials. In the macrophage stimulating assay using RAW 264.7 cells at 125~500 μg/mL of non-cytotoxicity doses, G-HW showed the most potent production of TNF-α, IL-6 and nitric oxide compared to red maca or the other extracts. The general component analysis results showed that all extracts comprised more than 90% neutral sugars with small amounts of uronic acid and protein. Meanwhile, component sugar analysis showed the difference in the content of uronic acids of cold- and hot-water extract. Additionally, the further fractionation of G-HW into crude polysaccharide (G-CP) greatly enhanced the macrophage stimulating activity, and G-CP contained macromolecules over 144 kDa, comprised mainly of glucose and galacturonic acid (51.0 and 34.9%). In conclusion, crude polysaccharide from maca showed industrial applicability as immunostimulating material, and especially golden maca showed higher macrophage stimulating activity than red maca.

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.

Suaeda asparagoides (Miq.) is a salt marsh plant, long been prescribed in traditional medicine for the treatment of hypertension and liver toxification in Asian countries. The powder of S. asparagoides was added at the ratio of 0, 5, and 10%, respectively, of grain-type Meju to manufacture Doenjang in brine according to the salt concentration (8 and 12%). After 24 weeks of fermentation, the Doenjang samples were determined to have an anti-inflammatory effect on RAW 264.7 cells. Evaluation of the anti-inflammatory effect of Doenjang added S. asparagoides powder extracted using 80% EtOH, was performed to study the inhibition of pro-inflammatory factors such as NF-κB (nuclar factor κB), NO (nitric oxide), TNF-α (tumor necrosis factor alpha), IL-6 (interleukin-6), iNOS (inducible nitric oxide synthase), and COX-2 (cyclooxygenase-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. The results showed that the Doenjang extracts reduced the production of NO, IL-6, COX-2, and iNOS increased in the LPS-stimulated RAW cell without cytotoxicity. In the case of the NF-κB and TNF-α there was no significant difference between the control and samples. In conclusion, these results suggest that Doenjang added with the S. asparagoides powder acts as functional fermented food with anti-inflammation effect.

Previous studies have suggested that rice bran oil (RBO), an edible oil from the byproducts of rice milling, has antiinflammatory effects in inflammation inducing macrophages, known as M1 subsets. Yet the effects of RBO on the counterpart M2 subsets, the “healing” macrophages, were poorly investigated to date. In this regard, recent studies on the molecular/cellular anti-inflammatory mechanisms of dietary components have demonstrated that mitochondrial respiration contributes to macrophage functioning. Therefore, the current study examined whether RBO regulates cytokine secretion by modulating mitochondrial metabolism in wound healing M2 subsets. Palm oil (PO), enriched with medium-chain fatty acids, served as a positive control. C57BL/6 mice were fed a diet containing either corn oil (CO), PO or RBO for 4 weeks, followed by purification of bone marrow-derived macrophages (BMDM) from their tibias and femurs. Cells were further polarized to M2-BMDM, and the expression of M2 marker (CD206) on cellular surfaces were not affected by dietary intervention. In addition, the secretion of anti-inflammatory cytokine (IL-10) in the culture supernatant was not affected by dietary lipids. Oxygen consumption rate, the indicator of mitochondrial respiration in M2-BMDM was not regulated by RBO intervention and PO treatment. Taken together, this study imply that RBO did not intervene both the regulation of inflammatory responses and mitochondrial respiration in M2 macrophages.

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.