• Archives of Pharmacal Research
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• 제27권2호
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• pp.217-224
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• 2004

Previously, we reported that water-extracted Acanthopanax senticasus exhibited anti-meta-static activity by stimulating the immune system. In this study, we fractionated glycoproteins (EN-SP) from the soluble protein layer (GF-AS) of A. senticasus and determined their basic chemical properties. We also investigated the anti-tumor and immunostimulating activities of the fractionated glycoprotein, EN-SP. We found that intravenous (i.v.) administration of GF-AS dramatically inhibited metastasis of colon26-M3.1 carcinoma cells to the lung in a dose-dependent manner. In vitro analysis showed GF-AS to enhance the proliferation of splenocytes. GF-AS also stimulated peritoneal macrophage, which was followed by the production of various cytokines such as IL-1$\beta$, TNF-$\alpha$, IL-12 and IFN-${\gamma}$. Furthermore, the production of these cytokines was partially blocked when peritoneal macrophage was cultured with the polyclonal antibodies against GF-AS. The depletion of NK cells by rabbit anti-asialo GM1 serum partly abolished the inhibitory effect of GF-AS on lung metastasis of colon26-M3.1 cells. Using gel filtration, EN-SP, an active glycoprotein fraction, is isolated from GF-AS. While both GF-AS and EN-SP stimulated the proliferatation of splenocytes of normal mice, EN-SP showed higher anti-metastatic activity and more potently stimulated the proliferation of splenocytes compared to GF-AS. These results suggest the use of EN-SP, the fractionated glycoprotein from A. senticasus, can be used as a therapeutical reagent to prevent or inhibit tumor metastasis.

• Mycobiology
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• 제38권2호
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• pp.144-148
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• 2010

$\beta$-Glucans have been known to exhibit antitumor activities by potentiating host immunity by an unknown mechanism. The C-type lectin dectin-1, a $\beta$-glucan receptor, is found on the macrophage and can recognize various $\beta$-glucans. Previously, we demonstrated the presence of $\beta$-glucan receptor, dectin-1, on the Raw 264.7 cells as well as on murine mucosal organs, such as the thymus, the lung, and the spleen. In order to investigate immunopotentiation of innate immunity by $\beta$-glucan, we stimulated a murine macrophage Raw 264.7 cell line with $\beta$-glucans from Pleurotus ostreatus, Saccharomyces cerevisiae, and Laminaria digitata. Then, we analyzed cytokines such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6 by reverse transcription-polymerase chain reaction (RT-PCR). In addition we analyzed gene expression patterns in $\beta$-glucan-treated Raw 264.7 cells by applying total mRNA to cDNA microarray to investigate the expression of 7,000 known genes. When stimulated with $\beta$-glucans, the macrophage cells increased TNF-$\alpha$ expression. When co-stimulation of the cells with $\beta$-glucan and lipopolysaccharide (LPS), a synergy effect was observed by increased TNF-$\alpha$ expression. In IL-6 expression, any of the $\beta$-glucans tested could not induce IL-6 expression by itself. However, when co-stimulation occurred with $\beta$-glucan and LPS, the cells showed strong synergistic effects by increased IL-6 expression. Chip analysis showed that $\beta$-glucan of P. ostreatus increased gene expressions of immunomodulating gene families such as kinases, lectin associated genes and TNF-related genes in the macrophage cell line. Induction of TNF receptor expression by FACS analysis was synergized only when co-stimulated with $\beta$-glucan and LPS, not with $\beta$-glucan alone. From these data, $\beta$-glucan increased expressions of immunomodulating genes and showed synergistic effect with LPS.

• 대한약침학회지
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• 제14권3호
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• pp.81-90
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• 2011

Objectives : Angelicae Gigantis Radix has been known traditional medicine with antimicrobial activities and it has been widely used for treatment of blood and inflammatory diseases. In the present study, some studies examined anti-inflammation effects of Angelicae Gigantis Radix but they usually were performed by ethanol extracted Angelicae Gigantis Radix pharmacopuncture. So We investigated the inhibitory effects of Angelicae Gigantis Radix pharmacopuncture by hot water and ethanol extract on Nitric oxide(NO) and Prostaglandin $E_2$($PGE_2$) production in lipopolysaccharide(LPS) induced macrophage cell. Methods : Angelicae Gigantis Radix was extracted by ethanol and hot water. Cell viability was determined by MTT assay. To evaluate anti-inflammation effects of Angelicae Gigantis Radix pharmacopuncture, we examined NO and $PGE_2$ production in LPS induced macrophages. The concentrations of NO and $PGE_2$ were measured by Griess assay and Enzyme Immuno-Assay. Results : 1) The MTT assay demonstrated that cytotoxic effect of Angelicae Gigantis Radix pharmacopuncture by hot water extract and ethanol extract in RAW 264.7 macrophage cells were not appeared. 2) Angelicae Gigantis Radix pharmacopuncture by ethanol extract and hot water extract inhibited NO production in LPS induced macrophages significantly. 3) Angelicae Gigantis Radix pharmacopuncture by ethanol extract tended to inhibiting $PGE_2$ production in LPS induced macrophages. And Angelicae Gigantis Radix pharmacopuncture by hot water extract inhibited LPS induced production of $PGE_2$ in RAW 264.7 macrophage cells significantly. Conclusions : This study suggests that Angelicae Gigantis Radix pharmacopuncture may have an anti-inflammatory property through the inhibition of NO and $PGE_2$ production in LPS induced macrophages. It may have a therapeutic potential for the treatment of various inflammatory diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제27권3호
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• pp.257-265
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• 2023

Kidney ischemia/reperfusion (I/R) injury, a common cause of acute kidney injury (AKI), is associated with the migration of inflammatory cells into the kidney. Ras-related C3 botulinum toxin substrate 1 (Rac1), a member of the Rho family of small GTPase, plays an important role in inflammatory cell migration by cytoskeleton rearrangement. Here, we investigated the role of Rac1 on kidney I/R injury and macrophage migration. Male mice were subjected to either 25 min of bilateral ischemia followed by reperfusion (I/R) or a sham operation. Some mice were administrated with either NSC23766, an inhibitor of Rac1, or 0.9% NaCl (vehicle). Kidney damage and Rac1 activity and expression were measured. The migration and lamellipodia formation of RAW264.7 cells, mouse monocyte/macrophage, induced by monocyte chemoattractant protein-1 (MCP-1, a chemokine) were determined using transwell migration assay and phalloidin staining, respectively. In sham-operated kidneys, Rac1 was expressed in tubular cells and interstitial cells. In I/R-injured kidneys, Rac1 expression was decreased in tubule cells in correlation with the damage of tubular cells, whereas Rac1 expression increased in the interstitium in correlation with an increased population of F4/80 cells, monocytes/macrophages. I/R increased Rac1 activity without changing total Rac1 expression in the whole kidney lysates. NSC23766 administration blocked Rac1 activation and protected the kidney against I/R-induced kidney damage and interstitial F4/80 cell increase. NSC23766 suppressed monocyte MCP-1-induced lamellipodia and filopodia formation and migration of RAW 264.7 cells. These results indicate Rac1 inhibition protects the kidney against I/R via inhibition of monocytes/macrophages migration into the kidney.

• Journal of Ginseng Research
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• 제48권2호
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• pp.181-189
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• 2024

Background: Cigarette smoke is generally accepted as a major contributor to chronic obstructive pulmonary disease (COPD), which is characterized by emphysematous lesions. In this study, we investigated the protective effects of Korean Red Ginseng (KRG) against cigarette smoke condensate (CSC)-induced emphysema. Methods: Mice were instilled with 50 mg/kg of CSC intranasally once a week for 4 weeks, KRG was administered to the mice once daily for 4 weeks at doses of 100 or 300 mg/kg, and dexamethasone (DEX, positive control) was administered to the mice once daily for 2 weeks at 3 mg/kg. Results: KRG markedly decreased the macrophage population in bronchoalveolar lavage fluid and reduced emphysematous lesions in the lung tissues. KRG suppressed CSC-induced apoptosis as revealed by terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling staining and Caspase 3 immunohistochemistry. Additionally, KRG effectively inhibited CSC-mediated activation of Bcl-2-associated X protein/Caspase 3 signaling, followed by the induction of cell survival signaling, including vascular endothelial growth factor/phosphoinositide 3-kinase/protein kinase B in vivo and in vitro. The DEX group also showed similar improved results in vivo and in vitro. Conclusion: Taken together, KRG effectively inhibits macrophage-mediated emphysema induced by CSC exposure, possibly via the suppression of pro-apoptotic signaling, which results in cell survival pathway activation. These findings suggest that KRG has therapeutic potential for the prevention of emphysema in COPD patients.

• Journal of Oral Medicine and Pain
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• 제20권1호
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• pp.53-65
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• 1995

The growth and synthetic activities of fibroblasts are regulated by cytokines and growth factors derived from activated inflammatory cells. Stimulatory effect of low level laser (LLL) radiation on wound healing seems to be in part due to direct stimulatory action on cell proliferation and synthetic activities of fibroblasts. Also indirect stimulatory effect on the fibroblast function through inflammatory or immune cells is another possible mechanism of biostimulatory action of LLL. This study was performed to determine the growth rate of human gingival fibroblasts obtained biopsy and culture, fibroblast cell line, and immune cell line by using $[^3H]-$ thymidine incorporation test. And gene expression pattern was also analyzed by using the DNA probe such as Hsp70, IL-1$\beta$, MIP-1$\alpha$ and actin cDNA. Proliferation rate of gingival fibroblast was increased by LLL irradiation, but no more effect was added by LPS or IL-1$\beta$ pretreatment Enhanced Hsp70 gene expression was found from gingival fibroblasts and fibroblast cell line COS by LLL irradiation., which was not more increased by LPS or IL-1$\beta$ pretreatment. LLL-irradiated promyelcytic cell line HL-60 and macrophage cell line RAW264.7 showed significant stimulatory effect of proliferation rate when compared with respective control. However there were no changes in growth rate of other immune cell tested in this study, such as B cell line WR19n.l and 230, helper T cell line Jurkat and Hut78, cytolytic T cell line CTLL-r8. By LLL-irradiation Hsp70 gene expression was increased in RAW246.7 and HL-60, not in CTLL-R8. And IL-1$\beta$ and MIP-1$\alpha$ gene expression were induced only from LLL-irradiated RAW264.7. These results led us to presume that LLL radiation may affect to the immune cells, especially to macrophage, through which it might promote wound healing process.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1145-1149
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• 2004

Immunomodulatory feed additives might offer alternatives to antimicrobial growth promoters in poultry production. This experiment was carried out to test the effect of $\beta$-glucan supplementation on the growth performance and immune response in broilers. Total of 160 day-old broilers were randomly assigned to 4 treatment groups fed corn-soybean diets containing 0, 0.012, 0.025 or 0.05% of $\beta$-glucan supplement in a 6 week feeding experiment. Growth performance, antibody titer against New Castle vaccine, lymphocyte blastogensis, and peritoneal macrophage chemotaxis activity of broilers were evaluated. Results showed that there were no significant differences in weight gain and feed efficiency among the treatments, and no differences in antibody titer was observed. Supplementation of $\beta$-glucan did not elevate the lymphocyte blastogensis among treatments, following stimulation with different mitogens. However, supplementation with 0.025 and 0.05% $\beta$-glucan enhanced the macrophage chemotaxis activity of broilers. These results suggest that $\beta$-glucan may enhance some cell-mediated immune responses of chickens by modulate macrophages ability.

• 한국자원식물학회지
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• 제33권3호
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• pp.183-193
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• 2020

Recently, as a natural substance has been emphasized interest in research to enhance the immune function. Green lettuce (Lactuca sativa L.) is a popular vegetable used fresh and it contains various phytochemicals and antioxidant compounds, and has been reported to have various physiological activities such as antibacterial, antioxidant, antitumor and anti-mutagenic. However, only a few studies have investigated on the mechanism of action of immune-enhancing activity of lettuce. Therefore, in this study, the immunomodulatory activities and potential mechanism of action of Green lettuce extracts (GLE) were evaluated in the murine macrophage cell line RAW264.7. GLE significantly increased NO levels by RAW264.7 cells, as well as expressions of immunomodulators such as iNOS, COX-2, IL-1β, IL-6, IL-12, TNF-α and MCP-1. Although GLE activated ERK1/2, p38, JNK and NF-κB, GLE-mediated expressions of immunomodulators was dependent on p38, JNK and NF-κB. In addition, TLR4 inhibition blocked GLE-mediated expressions of immunomodulators and activation of p38, JNK and NF-κB. Taken together, these results demonstrated that TLR4-MAPK/NF-κB signalling pathways participated in GLE-induced macrophage activation and GLE could be developed as a potential immunomodulating functional food.

• IMMUNE NETWORK
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• 제10권6호
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• pp.230-238
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• 2010

Background: The MK1 strain, a novel bacterial isolate from soft-rotten tissue of the Neungee mushroom, produces copious amounts of exopolysaccharide (EPS) in a dextrose minimal medium. This study examined the molecular characteristics and immunomodulatory activity of MK1 EPS. Methods: The EPS in the culture supernatant was purified by cold ethanol precipitation, and characterized by SDS- PAGE/silver staining and Bio-HPLC. The immunomodulatory activities of the EPS were examined using the mouse monocytic cell line, RAW 264.7 cells. Results: The molecular weights of the purified EPS were rather heterogeneous, ranging from 10.6 to 55 kDa. The EPS was composed of glucose, rhamnose, mannose, galactose, and glucosamine at an approximate molar ratio of 1.00 : 0.8 : 0.71 : 0.29 : 0.21. EPS activated the RAW cells to produce cytokines, such as TNF-${\alpha}$ and IL-$1{\beta}$, and nitric oxide (NO). EPS also induced the expression of co-stimulatory molecules, such as B7-1, B7-2 and ICAM-1, and increased the phagocytic activity. The macrophage-activating activity of EPS was not due to endotoxin contamination because the treatment of EPS with polymyin B did not reduce the macrophage-activating activity. Conclusion: The EPS produced from the MK1 strain exerts macrophage-activating activity.

• Journal of Nutrition and Health
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• 제49권5호
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• pp.288-294
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• 2016

Purpose: The aim of this work was to investigate the beneficial effects of rhamnazin against inflammation, reactive oxygen species (ROS)/reactive nitrogen species (RNS), and anti-oxidative activity in murine macrophage RAW264.7 cells. Methods: To examine the beneficial properties of rhamnazin on inflammation, ROS/ RNS, and anti-oxidative activity in the murine macrophage RAW264.7 cell model, several key markers, including COX and 5-LO activities, $NO^{\cdot}$, $ONOO^-$, total reactive species formation, lipid peroxidation, $^{\cdot}O_2$ levels, and catalase activity were estimated. Results: Results show that rhamnazin was protective against LPS-induced cytotoxicity in macrophage cells. The underlying action of rhamnazin might be through modulation of ROS/RNS and anti-oxidative activity through regulation of total reactive species production, lipid peroxidation, catalase activity, and $^{\cdot}O_2$, $NO^{\cdot}$, and $ONOO^{\cdot}$ levels. In addition, rhamnazin down-regulated the activities of pro-inflammatory COX and 5-LO. Conclusion: The plausible action by which rhamnazin renders its protective effects in macrophage cells is likely due to its capability to regulate LPS-induced inflammation, ROS/ RNS, and anti-oxidative activity.