• Journal of Haehwa Medicine
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• v.9 no.1
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• pp.215-223
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• 2000

To understand immunopharmacologic effects on Cervi Pantotrichum Cornu, Morindae Officinalis Radix, Cistanches Herba, Curculginis Rhizoma, Epimedii Herba, Eucommiae Cortex, we investigated chinese experimental documents, and we could reach conclusions as follows : 1. The effects on cell-mediated immune system were as follows. 1) The effects on macrophage (1) The herbal medicines promoting to increase the number of WBC in the peripheral blood were Morindae Officinalis Radix, Epimedii Herba and that promoting to reinforce the phagocytic functions of neutrophil was Curculginis Rhizoma. (2) The herbal medicines promoting the phagocytic functions of mononuclear, macrophage were Cervi Pantotrichum Cornu, Cistanches Herba, Eucommiae Cortex. 2) The herbal medicines stimulating the activities of T lymphocytes were Cervi Pantotrichum Cornu, Curculginis Rhizoma, Epimedii Herba, Eucommiae Cortex. 2. The effects on humoral immune system were as follows. 1) The herbal medicines increasing the activity of complement receptor were Cervi Pantotrichum Cornu, Curculginis Rhizoma. 2) The herbal medicines reinforcing immunity of spleen cells were Cervi Pantotrichum Cornu, Cistanches Herba, Epimedii Herba. 3) The herbal medicines promoting proliferation of spleen cells that produce antibody after having been immunized by SRBC were Cervi Pantotrichum Cornu, Cistanches Herba, Epimedii Herba. 3. The herbal medicines, reinforcing immunity on delayed type hypersensitivity were Cervi Pantotrichum Cornu, Cistanches Herba, Eucommiae Cortex. As you know in the many bibliological documents, the studies on the effects of Drugs for Tonifying Yang were started along right lines. Recently the studies on those were accomplished more rapidly and applied many immune diseases. We thought that Drugs for Tonifying Yang could be important immunopotentiators. Therefore we can apply those herbal medicines not only to immune diseases but also inflammatory diseases, senile infirmity and all sorts of tumor.

• The Korea Journal of Herbology
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• v.37 no.5
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• pp.53-61
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• 2022

Objectives : The aim of this study was to investigate the effect of Baicalein (BA) on the production of hydrogen peroxide in lipoteichoic acid-stimulated RAW 264.7 mouse macrophages. Methods : Lipoteichoic acid-stressed RAW 264.7 mouse macrophages were incubated with baicalein at concentrations of 50 and 100 µM. Incubation time is 30 minutes, 2 h, 12 h, and 18 h. After incubation, The production of hydrogen peroxide in RAW 264.7 mouse macrophages was measured with dihydrorhodamine 123 assay. Streptococcus aureus lipoteichoic acid and Streptococcus pyogenes lipoteichoic acid were used as cell-stimulating lipoteichoic acid. Cell viabilities were measured with a modified MTT assay. Berberine, indomethacin, and gallic acid were incubated for the same time as the comparative materials. Results : BA at the concentration of 50 and 100 µM did not show cytotoxicity on RAW 264.7 mouse macrophages for 24 h incubation. For 30 minutes, 2 h, 12 h, and 18 h incubation, BA at the concentration of 50 and 100 µM significantly inhibited the production of hydrogen peroxide in RAW 264.7 mouse macrophages stimulated by Streptococcus aureus lipoteichoic acid (p < 0.05); also, BA at the concentration of 50 and 100 µM also inhibited the productions of hydrogen peroxide in RAW 264.7 mouse macrophages stimulated by Streptococcus pyogenes lipoteichoic acid significantly (p < 0.05). Conclusions : BA might have anti-bacterial activity related to its inhibition of hydrogen peroxide production in lipoteichoic acid-stimulated RAW 264.7 mouse macrophages.

• Journal of Microbiology
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• v.45 no.4
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• pp.305-310
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• 2007

The principal objective of this study was to compare the effects of whole and hydrolyzed cells (bifidobacteria) treated with gastrointestinal digestive enzymes on the activation of cloned macrophages. Seven different strains of Bifidobacterium obtained from swine, chickens, and rats, were digested with pepsin followed by pancreatin and the precipitate (insoluble fraction) and supernatant (soluble fraction) obtained via centrifugation. The RAW 264.7 murine macrophages were incubated with either whole cells, the precipitate, or supernatant at various concentrations. Pronounced increases in the levels of nitric oxide (NO), interleukin $(IL)-1{\beta}$, IL-6, IL-12, and tumor necrosis factor $(TNF)-{\alpha}$ were observed in the whole cells and precipitates, but these effects were less profound in the supernatants. The precipitates also evidenced a slight, but significant, inductive activity for NO and all tested cytokines, with the exception of $(TNF)-{\alpha}$ in the macrophage model as compared with the whole cells. By way of contrast, $(TNF)-{\alpha}$ production when cultured with whole cells (100 ng/ml) resulted in marked increases as compared with what was observed with the precipitates. The results of this study indicated, for the first time, that digested Bifidobacterium sp. can induce the production of NO and several cytokines in RAW 264.7 murine macrophage cells. In the current study, it was demonstrated that Bifidobacterium strains treated with digestive enzymes, as compared with whole cells, are capable of stimulating the induction of macrophage mediators, which reflects that they may be able to modulate the gastrointestinal immune functions of the host.

• KSBB Journal
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• v.23 no.2
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• pp.164-168
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• 2008

The purpose of this study was to confirm the content of total polyphenol, antioxidative and immune activities of the extracts of Cedrela sinensis leaf. The content of total polyphenol of water extracts ranged from 46.5-59.6 mg/100 g, which was higher than other extracts using organic solvents such as EtOAc, $CH_2Cl_2$ and $C_6H_{14}$. The antioxidant activity of the water and organic solvents extracts showed 6-33% in terms of 2,2-diphenyl-picryl-hydrazyl (DPPH) scavenging activity. To analyze the immuno-stimulation activity of C. sinensis leaf extract, we investigated the effect of the extracts on NO synthesis which is important in host defense against bacterial infection. Hot water extracts significantly increased NO generation by RAW 264.7, macrophage cell line, while organic solvent extract has no significant effect on NO production. To further analyzed the anti-inflammatory effect of the extracts, we investigated the effects of the extracts on lipopolysaccharide(LPS)-induced NO generation. Organic solvent extracts of C. sinensis leaves showed strong inhibitory effect on NO production in LPS-stimulated RAW 264.7 cells. These results suggest that C. sinensis leaf extract may represent a useful immune stimulating agent and anti-inflammatory agent.

• Biomedical Science Letters
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• v.13 no.3
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• pp.161-168
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• 2007

Salicornia herbacea L. (Chenopodiaceae: S. herbacea) is a salt marsh plant, which has long been prescribed in traditional medicines for the treatment of intestinal ailments, nephropathy, and hepatitis in Oriental countries. In order to elucidate the mechanisms of this herb, we conducted an anti-oxidative activity, the inhibition of nitric oxide (NO) production, and the suppression of the pro-inflammatory cytokine genes, with the solvent-extracts of S. herbacea. We found that both ethyl acetate and n-butanol tractions showed potent anti-oxidative effects in comparison to other fractions using xanthine oxidase assay with $IC_{50}$ values of $66.0{\pm}0.5\;{\mu}g/ml$ and $82.5{\pm}3.8\;{\mu}g/ml$, respectively. In addition, both ethyl acetate and n-butanol fractions showed more electron donating activity (EDA) than other tractions, according to DPPH (2, 2-Diphenyl-lpicrylhydrazyl radical) assay. The EDA of ethyl acetate fraction ($IC_{50}$ values of $117.5{\pm}3.8\;{\mu}g/ml$) is more significant than that of n-butanol fraction ($IC_{50}$ values of $375.0{\pm}12.5\;{\mu}g/ml$). Among potential anti-oxidative tractions, ethyl acetate traction dose-dependently suppressed lipopolysaccharide (LPS, $0.1\;{\mu}g/ml$)-induced nitric oxide (NO) production in RAW264.7 cell, while n-butanol did not. As expected, ethyl acetate fraction suppressed the expression of inducible NO synthase (iNOS) in RAW264.7 cell stimulated by $0.1\;{\mu}g/ml$ of LPS. Moreover, the ethyl acetate traction suppressed the expression of interleukin-1 $(IL)-1{\beta}$ and granulocyte/macrophage colony-stimulating factor (GM-CSF) mRNA in LPS-stimulated RAW264.7 cells. Therefore, these results suggest that S. herbacea may have anti-oxidative and anti-inflammatory activities by modulating radical-induced toxicity and various pro-inflammatory responses.

• The Journal of Korean Obstetrics and Gynecology
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• v.26 no.4
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• pp.66-81
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• 2013

Purpose: This study was carried out to investigate the anti-inflammatory effects of Ligustri Lucidi Fructus water extract (LF) in the lipopolysaccharide (LPS)-induced mouse macrophages RAW 264.7 cell. Methods: Ligustri Lucidi Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of LF, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. To investigate anti-inflammatory effects of LF, the concentration of nitric oxide (NO) was measured with NO assay, cytokine was measured by Bio-Plex cytokine assay, and intracellular calcium (Ca) was measured with Fluo-4 Ca assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically (P<0.05). Results: 1. LF showed no cytotoxicity. 2. LF inhibited significantly the production of NO at the concentration of 25, 50 and $100{\mu}g/ml$. 3. LF inhibited significantly the production of interleukin (IL)-4, macrophage inflammatory protein (MIP)-$1{\alpha}$, granulocyte colony stimulating factor (G-CSF) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. LF inhibited significantly the production of granulocyte macrophage-colony stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF) at the concentration of 50, 100 and $200{\mu}g/ml$, the interferon (IFN)-${\gamma}$ at 25, 50 and $100{\mu}g/ml$ respectively. 5. LF inhibited significantly the production of IL-$1{\beta}$ at the concentration of 50 and $200{\mu}g/ml$, the IL-5 at 25 and $100{\mu}g/ml$, the IL-12p70, MIP-$1{\beta}$ at 50 and $100{\mu}g/ml$, the regulated on activation, normal T cell expressed and secrete d (RANTES) at 100 and $200{\mu}g/ml$ respectively. 6. LF inhibited significantly the production of IL-10, interferon gamma-induced protein (IP)-10 at the concentration of $200{\mu}g/ml$. 7. LF inhibited significantly the production of intracellular Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. Conclusions: These results suggest that LF has anti-inflammatory effect and immuno-modulating activity.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.329-333
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• 2003

Human granulocyte-macrophage colony-stimulating factor (hGM-CSF) is a glycoprotein that stimulates the production of granulocytes, macrophages and white blood cells. hGM-CSF secreted by transgenic Nicotiana tabacum suspension cells was unstable in the culture medium and rapidly degraded by extracellular preteases. In order to reduce extracellular pretense activity, culture temperature was lowered. Then, the production of hGM-CSF by transgenic plant suspension cell cultures could be enhanced by reduced degradation of hGM-CSF at low temperature.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.341-345
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• 2003

Effect of immobilization on the production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) by Nicotiana tabacum cells was investigated using polyurethane foam as immobilization matrices. The cell activity and the hGM-CSF production were maintained for 16 days in spite of 3 times of media exchange. Under the same conditions of temperature and agitation rate, maximum concentrations of hGM-CSF in a 500-mL spinner flask and 100-mL Erleuneyer flasks were 17.3 ${\mu}g/L$ and 9.8 ${\mu}g/L$, respectively. Consequently high hGM-CSF production could be possible in spinner flask when the rate and amount of media exchange were optimized.

• Food Science and Biotechnology
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• v.14 no.1
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• pp.147-151
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• 2005

Purified compound with intestinal immune system-modulating properties, GWE-2c, was isolated from methanol extract of Gelidium amansii by sequential procedures with silica gel column, LH-20 Sephadex gel column, and thin-layer chromatographies. In the presence of GWE-2c, strong immunoactivity in Peyers patch cell-mediated bone marrow cells was observed in vitro. In vivo intestinal immune-modulating activity was also enhanced by crude phenolic compound (GWE) of G. amansii in a dose-dependent manner. Investigation of production of several cytokines in Peyer's patch cells upon stimulation with GWE in vivo revealed the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-6 increased. Results suggest that the phenolic compound from G. amansii represents immunopotentiator and biological response modifier at in vitro and in vivo levels.