• Korean Journal of Plant Resources
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• v.34 no.6
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• pp.536-541
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• 2021

In this study, we investigated whether Hibiscus manihot flowers (HMF) exhibits immunostimulatory activity in RAW264.7 cells. HMF increased the production of immunostimulatory factors such as NO, iNOS, IL-1β and TNF-α in RAW264.7 cells. TLR2 and TLR4 blocked HMF-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of MAPK signaling pathway reduced HMF-mediated production of immunostimulatory factors. From these results, HMF is thought to promote the production of immunostimulatory factors through activating TLR2/4/MAPK signaling in macrophages. It is believed that HMF can be developed as an agent related to immune enhancement in the future.

• Korean Journal of Plant Resources
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• v.34 no.4
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• pp.377-383
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• 2021

In this study, we investigated in vitro immunostimulatory activity of fruit extracts from Pyrus ussuriensis var. hakunensis (Nakai) T.B. Lee (PUF) using mouse macrophage RAW264.7 cells. PUF increased the production of immunostimulatory factors such as NO, iNOS, IL-1β, IL-6 and TNF-α, and phagocytic activity in RAW264.7 cells. The inhibition of TLR2 and TLR4 blocked PUF-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of MAPKs signaling pathway reduced PUF-mediated production of immunostimulatory factors. From these results, PUF may have immunostimulatory activity via TLR2/4-mediated activation of MAPKs signaling pathway. Therefore, PUF expected to be used as a potential immune-enhancing agent.

• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.934-940
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• 2023

Syneilesis palmata (SP) is a traditional medicinal plant. SP has been reported to have anti-inflammatory, anticancer, and anti-human immunodeficiency virus (HIV) activities. However, there is currently no research available on the immunostimulatory activity of SP. Therefore, in this study, we report that S. palmata leaves (SPL) activate macrophages. Increased secretion of both immunostimulatory mediators and phagocytic activity was observed in SPL-treated RAW264.7 cells. However, this effect was reversed by the inhibition of TLR2/4. In addition, inhibition of p38 decreased the secretion of immunostimulatory mediators induced by SPL, and inhibition of TLR2/4 decreased the phosphorylation of p38 induced by SPL. SPL augmented p62/SQSTM1 and LC3-II expression. The increase in protein levels of p62/SQSTM1 and LC3-II induced by SPL was decreased by the inhibition of TLR2/4. The results obtained from this study suggest that SPL activates macrophages via TLR2/4-dependent p38 activation and induces autophagy in macrophages via TLR2/4 stimulation.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.87-87
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• 2022

Under the COVID-19 pandemic, interest in immune enhancement and anti-obesity is increasing. Thus, in this study, we investigated whether Kadsura japonica fruits (KJF) exhibits immunostimulatory activity and anti-obesity activity. KJF increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. Inhibition of TLR2 and TLR4 blocked KJF-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of MAPK and PI3K/AKT signaling pathway reduced KJF-mediated production of immunostimulatory factors, and the activation of MAPK and PI3K/AKT signaling pathway by KJF suppressed the inhibition of TLR2/4. KJF attenuated the lipid accumulation and the protein expression such as CEBPα, PPARγ, perilipin-1, adiponectin, and FABP4 related to the lipid accumulation in 3T3-L1 cells. In addition, KJF inhibited excessive proliferation of 3T3-L1 cells and protein expressions such as β-catenin and cyclin D1 related to cell growth. These findings indicate that KJF may have immunostimulatory activity and anti-obesity activity.

• The Journal of Korean Medicine
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• v.41 no.4
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• pp.52-63
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• 2020

Objectives: In this study, we investigated the effects of Pueraria lobata Ohwi flower extracts (PLFE) on macrophages and their underlying mechanism(s) of action. PLFE increased the production of NO and cytokines (IL-6 and TNF-𝛼) in a dose-dependent manner, indicating its immunostimulatory property. Furthermore, PLFE upregulated iNOS, COX-2, and mitogen-activated protein kinase (MAPK) signaling in RAW264.7 cells. Additionally, PLFE enhanced the phosphorylation of I𝜅B𝛼 and subsequent I𝜅B𝛼 degradation, thereby enabling the nuclear translocation of NF-𝜅B. Taken together, these findings demonstrate that the immunostimulatory effects of PLFE are mediated by the nuclear translocation of the p65 subunit of NF-𝜅B and subsequent secretion of cytokines (IL-6 and TNF-𝛼), upregulation of iNOS and COX-2, and stimulation of MAPK signaling (JNK, ERK, and p38). Thus, PLFE may be a potential immunostimulatory therapeutic.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2022.09a
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• pp.88-88
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• 2022

In this study, we investigated the effect of Solanum nigrum aerial parts (SNAP) on macrophage activation and macrophage autophagy in RAW264.7 cells. SNAP increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. TLR4 inhibition blocked SNAP-mediated production of immunostimulatory factors. In addition, the JNK inhibition reduced the SNAP-mediated production of immunostimulatory factors, and the SNAP-mediated JNK activation was blocked by the TLR4 inhibition. SNAP activated macrophage autophagy. TLR4 inhibition blocked SNAP-mediated macrophage autophagy and inhibition of p38 and JNK attenuated SNAP-mediated macrophage autophagy. These findings indicate that SNAP may induce TLR4/JNK-mediated macrophage activation and TLR4/p38 and JNK-mediated macrophage autophagy.

• Journal of Food Hygiene and Safety
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• v.31 no.2
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• pp.126-131
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• 2016

This study examined the immunostimulatory activities on a 70% ethanol extract of raw garlic (RG) and steamed garlic (SG) grown in Korea, Japan, and America using primary immune cells isolated from the blood of Soyangin, Taeumin, and Soeumin volunteers. The Soeumin showed the highest cell proliferation, NO and $TNF-{\alpha}$ production from the immune cells treated with a 70% ethanol raw garlic extract (RGE), followed Taeumin, and Soyangin. The constitution differences were not observed for the same ethanol streamed garlic extract (SGE) and the SGE showed stronger immunostimulatory activities than RGE. The immunostimulatory activities of RG and SG showed a significant difference among countries and the immunostimulatory activities were the highest in RG and SG grown in Korea, followed by RG and SG grown in Japan, RG and SG grown in America. The effect of RGE and SGE on antioxidant activity showed similar trends.

• Food Science of Animal Resources
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• v.42 no.6
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• pp.1031-1045
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• 2022

Postbiotics are defined as probiotics inactivated by heat, ultraviolet radiation, sonication, and other physical or chemical stresses. Postbiotics are more stable than probiotics, and these properties are advantageous for food additives and pharmacological agents. This study investigated the immunostimulatory effects of heat-treated Lactiplantibacillus plantarum LM1004 (HT-LM1004). Cellular fatty acid composition of L. plantarum LM1004 isolated form kimchi was analyzed by gas chromatography-mass spectrometry detection system. The nitric oxide (NO) content was estimated using Griess reagent. Immunostimulatory cytokines were evaluated using enzyme-linked immunosorbent assay. Relative protein expressions were evaluated by western blotting. Phagocytosis was measured using enzyme-labelled Escherichia coli particles. L. plantarum LM1004 showed 7 kinds of cellular fatty acids including palmitic acid (C16:0). The HT-LM1004 induced release of NO and upregulated the inducible NO synthase in RAW 264.7 macrophage cells. Tumor necrosis factor-α and interleukin-6 levels were also increased compared to control (non-treated macrophages). Furthermore, HT-LM1004 modulated mitogen-activated protein kinase (MAPK) subfamilies including p38 MAPK, extracellular signal-regulated kinase 1/2, and c-Jun N-terminal kinase. Therefore, these immunostimulatory effects were attributed to the production of transcriptional factors, such as nuclear factor kappa B (NF-κB) and the activator protein 1 family (AP-1). However, HT-LM1004 did not showed significant phagocytosis of RAW 264.7 macrophage cells. Overall, HT-LM1004 stimulated MAPK/AP-1 and NF-κB expression, resulting in the release of NO and cytokines. These results will contribute to the development of diverse types of food and pharmacological products for immunostimulatory agents with postbiotics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.9
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• pp.1325-1333
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• 2014

Sasang constitutional medicine is a Korean traditional medicine in which individuals are classified into four constitutional types, Taeyangin, Taeumin, Soyangin, and Soeumin. To develop new functional Saengsik products, we investigated the immunostimulatory activities of raw materials of Saengsik (milled rice, barley, Job's tears, soybean, adzuki bean, millet, eggplants, radish, carrot, Lentinus edodes, Agaricus bisporus, and Auricularia auricula-judae) and Saengsik products (Mix 1~6) in primary immune cells (lymphocytes and macrophages) isolated from blood of Taeumin, Soyangin, and Soeumin types. Increased cell proliferation as well as NO and TNF-${\alpha}$ protein production by immune cells treated with extracts of materials of Saengsik and Saengsik products were measured as immunostimulatory parameters. We chose milled rice, adzuki bean, radish, and Lentinus edodes to study the immunostimulatory activity of Teaumin, barley, soybean, eggplants, and Agaricus bisporus to study the immunostimulatory activity of Soyangin, as well as Job's tears, adzuki bean, carrot, and Auricularia auricula-judae to study the immunostimulatory activity of Soeumin. The mixtures (Mix 1~6) were made with different kinds and ratios of Saengsik materials based on the immunostimulatory activities of Saengsik materials. The immunostimulatory activity of Taeumin was highest in Mix 2 (rice 60%, adzuki bean 20%, radish 10%, and Lentinus edodes 10%) extract-treated immune cells among all six mixtures. The immunostimulatory activity of Soyangin responded best to Mix 4 (barley 60%, soybean 20%, eggplants 10%, and Agaricus bisporus 10%) among the six extracts (Mix 1~6). The increased immunostimulatory activity of Mix 6 (Job's tears 60%, adzuki bean 20%, carrot 10%, and Auricularia auricula-judae 10%)-treated immune cells was higher than the other five extracts (Mix 1~5). Accordingly, Mix 2, Mix 4, and Mix 6 may be useful as mixtures for Saengsik products having personalized immunostimulatory activities.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.44-44
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• 2023

Syneilesis palmata (SP) has been used as a traditional medicinal plant and vegetable. SP was reported to exert pharmacological activities such as anti-inflammation, anti-cancer, and anti-HIV. However, there are no studies on the immunostimulatory activity of SP. Thus, in this study, we report that S. palmata leaves (SPL) induce the activation of macrophages. An increase in both secretions of immunostimulatory mediators and phagocytotic activity was observed in SPL-treated RAW264.7 cells. However, this was reversed by inhibition of TLR2/4. In addition, the p38 inhibition reduced the SPL-mediated secretion of immunostimulatory mediators, and the SPL-mediated p38 activation was blocked by the TLR2/4 inhibition. SPL augmented both p62/SQSTM1 and LC3-II. TLR2/4 inhibition blocked the SPL-mediated increase of p62/SQSTM1 and LC3-II. These findings indicate that SPL may activate macrophages through TLR2/4-dependent p38 activation and activate autophagy through TLR2/4 stimulation.