• IMMUNE NETWORK
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• v.3 no.1
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• pp.38-46
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• 2003

Background: Platelet-activating factor (PAF) induces nuclear factor $(NF)-{\kappa}B$ activation and angiogenesis and increases tumor growth and pulmonary tumor metastasis in vivo. The role of $NF-{\kappa}B$ activation in PAF-induced angiogenesis in a mouse model of Matrigel implantation, and in PAF-mediated pulmonary tumor metastasis were investigated. Methods: Angiogenesis using Matrigel and experimental pulmonary tumor metastasis were tested in a mouse model. Electrophoretic mobility shift assay was done for the assessment of $NF-{\kappa}B$ translocation to the nucleus. Expression of angiogenic factors, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\alpha}$, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were tested by RT-PCR and ELISA. Results: PAF induced a dose- and time-dependent angiogenic response. PAF-induced angiogenesis was significantly blocked by PAF antagonist, CV6209, and inhibitors of $NF-{\kappa}B$ expression or action, including antisense oligonucleotides to p65 subunit of $NF-{\kappa}B$ (p65 AS) and antioxidants such as ${\alpha}$-tocopherol and N-acetyl-L-cysteine. In vitro, PAF activated the transcription factor, $NF-{\kappa}B$ and induced mRNA expression of $TNF-{\alpha}$, $IL-1{\alpha}$, bFGF, VEGF, and its receptor, KDR. The PAF-induced expression of the above mentioned factors was inhibited by p65 AS or antioxidants. Also, protein synthesis of VEGF was increased by PAF and inhibited by p65 AS or antioxidants. The angiogenic effect of PAF was blocked when anti-VEGF antibodies was treated or antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF was co-administrated, but not by antibodies against $TNF-{\alpha}$, $IL-1{\alpha}$, and bFGF each alone. PAF-augmented pulmonary tumor metastasis was inhibited by p65 AS or antioxidants. Conclusion: These data indicate that PAF increases angiogenesis and pulmonary tumor metastasis through $NF-{\kappa}B$ activation and expression of $NF-{\kappa}B$-dependent angiogenic factors.

• Journal of Life Science
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• v.22 no.6
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• pp.828-836
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• 2012

The objective of the current study was to determine the effects of the extracts isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants on mouse splenocytes, B cells, and macrophages in vitro. The ethanol extracts B (EEB) directly induced the proliferation of spleen cells in a dose-dependent manner and increased IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ synthesis. The EEB also increased the proliferation of B cells in a dose-dependent manner. The production of immunoglobulin M, G1, G2a, G2b, and IgG3 in the presence of the EEB increased progressively in the culture supernatant. When the EEB were used in macrophage cell line (RAW264.7) stimulation, there was a marked induction of NO synthesis in a dose-dependent manner and an increased IL-6, TNF-${\alpha}$, and GM-CSF synthesis. Intraperitoneal injection with EBB showed life prolongation effect of 16.1% in mice previously inoculated with sarcoma-180, respectively. These results suggest that the capacity of the EEB isolated from the fruit body of Flammulina velutipes cultivated with oriental herbal plants seems to act as a potent immunomodulator causing augmentation of immune cell activity, and with the absence of notable side-effects, Flammulina velutipes EEB could be used as a biological response modifier having possible therapeutic effects against immunological disorders. This study also showed that functional components of Flammulina velutipes were possibly improved by incorporating oriental herbal plants in a growth medium.

• Environmental Analysis Health and Toxicology
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• v.22 no.1 s.56
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• pp.65-71
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• 2007

유산균(Lactic acid bacteria)은 Escherichia coli와 Salmonella typhimurium과 같은 병원균에 대한 항균활성을 나타낼 뿐만 아니라 면역 증강효과를 나타내는 등 인체내에서 건강에 이로운 다양한 역할을 수행하는 것으로 알려졌다. 특히, Pediococcus pentosaseus와 몇몇 Lactobacillus 종으로부터 분리한 항균활성을 나타내는 peptide들인 safelac과 lactopad는 몇몇 암세포주의 성장을 억제하는 것으로 나타났다. 이에, 본 연구에서는 HT-29, SW 480 및 Caco-2와 같은 3종류의 인간의 결장암 세포주에 safelac과 lactopad를 투여하여 이들이 항암효과를 나타낼 수 있는 지를 분석하고자 하였다. XTT assay는 safelaf과 lactopad가 HT-29, SW 480 및 Caco-2의 성장을 억제하는 것으로 나타났으며, 특히, 이들 peptide들을 72시간동안 처리했을 때 나타나는 항암효과는 $3.1{\sim}100mg/mL$의 농도범위에서 유의한 결과를 나타내었으며, 분석한 농도 범위에서 용량 의존적인 방식으로 더 강한 효과를 나타내었다. RAW 264.7 세포주는 cytokine인 tumor-necrosis factor(TNF-${\alpha}$)의 생성에 미치는 이들 peptide들의 효과를 조사하기 위한 대식세포의 모델로써 이용되었다. RAW 264.7 세포주에서 TNF-${\alpha}$의 생성은 이들 peptide들에 의해 48시간 배양시 용량에 의존적인 방식으로 영향을 받는 것으로 나타났다. 따라서, 이러한 발견은 safelac과 lactopad와 같은 유산균으로부터 분리한 항균 peptide들이 결장암 세포에 대한 화학적 예방제로서의 잠재성을 갖고 있음을 시사하는 결과로서 주목된다.

• The Journal of Korean Medicine
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• v.42 no.3
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• pp.56-71
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• 2021

Objectives: This study is aimed to evaluate the protective effects of GGX on lung injury of Chronic Obstructive Lung Disease (COPD) mice model. Materials and Methods: C57BL/6 mice were challenged with lipopolysaccharide (LPS) and cigarette smoke extract (CSE) and then treated with vehicle only (Control group), dexamethasone 3 mg/kg (Dexa group), gam-gil-tang 200 mg/kg (GGT group), GGX 100, 200, and 400 mg/kg (GGX group). After sacrifice, its bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed with cytospin, Enzyme-Linked Immunosorbent Assay (ELISA), real-time polymerase chain reaction (PCR) and hematoxylin & eosin (H&E), and Masson's trichrome staining. Results: In the COPD model, GGX significantly inhibited the increase of neutrophils, TNF-𝛼, IL-17A, CXCL-1, MIP2 in BALF and TNF-𝛼, IL-1𝛽, IL-10 mRNA expression in lung tissue. It also decreased the severity of histological lung injury. Conclusion: This study suggests the usability of GGX for COPD patients by controlling lung tissue injury.

• Journal of Acupuncture Research
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• v.24 no.4
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• pp.167-181
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• 2007

Objectives: To evaluate expression of MIF mRNA, MIF, $TNF-{\alpha}$, $IL-6R-{\alpha}$, STAT-3, $NF-{\kappa}B$ p65, COX-2 and iNOS, MMP-9mRNA after injecting deer antler herbal acupuncture solution in a LPS rat model. Methods: The experiment was divided in category of the control group, RA group, and NA group. RA was induced in the mice via injecting 300ug/kg LPS. The deer antler herbal acupuncture solution 50ug/kg was applied on $ST_{35}$(犢鼻) and EX-LE201(內膝眼) for 19days from $3^{rd}$ day of RA inducement. Results: 1. In the deer antler herbal acupuncture solution treated RAW 264.7cell, the mRNA expression of cytokines, RA related inflammation factors, such as the MIF, COX- 2, iNOS, and MMP-g reduced concentration dependently. 2. In the deer antler herbal acupuncture treated mice's synovial membrane, decrease in the cell replication of synovial joint cells, regeneration of blood vessel, fibrosis and fibroblastic cells expansion were observed. 3. Positive reaction of RA-related cytokines MIF, $TNF-{\alpha}$, $IL-6R-{\alpha}$, STAT3, COX-2, iNOS, $NF-\;{\kappa}B$ p65, MMP-9 was reduced. Conclusion : On the basis of the results, it was concluded that deer antler herbal acupuncture extract has significant protecting ability against acute progressive RA by inhibiting the production of MIF, as a top in cytokines related to inflammation.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.10a
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• pp.340-344
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• 2004

젖소 초유 중에 함유된 IGF-I rich fraction을 30kDa와 1kDa ultrafiltration(UF) membrane을 이용하여 효과적으로 분리하였으며 분획내의 IGF-I은 SDS-PAGE와 Western blot으로 확인하였다. 분획한 IGF-I rich fraction이 murine macrophage의 면역 활성에 미치는 영향을 실험한 결과 $1{\mu}g$ 투여군의 경우 IL-6는 7.3ng/ml, NO는 $10.7{\mu}M$을 생성하였고 Phagocytosis는 대조구 대비하여 65% 증진시켰다. $TNF-{\alpha}$의 분비량은 L929세포의 증식 저해율로 측정한 결과 대조구에 대비하여 30% 더 높은 저해율을 나타내었고, $H_2O_2$는 대조구에 대비하여 6% 더 높은 과산화수소를 생산하였다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.107-107
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• 2002

ADAM은 metalloprotease/disintegrin domain을 가진 transmembrane glycoprotein 으로서 지금까지 30개 이상의 ADAM 및 10개 이상의 ADAMTS 단백질이 알려져 있다. 이들의 기능은 포유동물의 수정시 sperm-egg binding과 fusion, myoblast fusion, integrin과의 결합 등에 직접 관여하거나, TNF-alpha 등의 생체 신호전달물질이 세포로부터 분비될 때에 이들의 구조를 변화시켜 활성화시키는 효소작용, 그리고 dendritic cell differentiation 등에 관여하는 것으로 알려져있다. 그러나 자궁내막 조직에서의 유전자 및 단백질 발현 여부에 관해서는 거의 보고되어 있지 않고 있다. 먼저 RT-PCR 방법을 이용하여 자궁에서 mRNA의 양을 $\beta$-actin의 양에 대하여 상대적으로 측정한 결과, 임신 1일부터 5일까지는 ADAM-9, 10, 17, TS1의 mRNA 경우 거의 비슷하게 발현되었으며 ADAM-8과 15는 3일째, ADAM-12는 2일째와 4일째에 현저하게 증가하였다 임신 6일부터 8일까지의 자궁조직을 각각 embryo가 착상된 곳과 그렇지 않은 곳으로 나누어 조사한 결과 상대적으로 착상이 된 곳에서 mRNA가 많이 발현 되었으며 특히 ADAM-8, 12, 15의 mRNA의 경우 현저한 차이를 보였다. Immunoblotting 방법을 이용하여 임신 1일부터 5일까지의 단백질의 발현 양상을 조사한 결과 임신 ADAM-8은 3일 이후 감소되는 양상을 보였으며 ADAM-9, 15, TS1은 5일째에 증가하는 양상을 보였다. ADAM-10은 2일째 감소하다가 다시 증가하고 ADAM-12, 17은 3, 4일째 증가하다가 5일째 감소하는 상을 보였다. 한편 임신 6일부터 8일까지는 embryo가 착상된 곳이 그렇지 않은 곳보다 조사된 모든 ADAMs 단백질이 상대적으로 많이 발현되는 양상을 보였다. 이러한 결과로 미루어 ADAMs은 임신 초기 착상과정과 임신 단계에 따른 자궁의 조직 재구성에 중요한 역할을 할 것으로 생각된다.

• Korean Journal of Plant Resources
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• v.31 no.4
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• pp.294-302
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• 2018

This study was conducted to compare anti-inflammatory effect of Robinia pseudoacacia L. using different extraction methods (water extraction, ethanol extraction and high temperature extraction). We investigated anti-inflammatory effect of Robinia pseudoacacia L. extract (RP1, water extract; RP2, ethanol extract; RP3, high temperature extract) on lipopolysaccharide (LPS)-stimulated inflammation using Raw 264.7 cell. Cells were treated with various concentrations (12.5, 25, 50, 100 or $200{\mu}g/m{\ell}$) of water extract, ethanol extract and high temperature extract. Cytotoxicity was not observed on Raw 264.7 cells, LPS-stimulated production of NO (nitric oxide), $PGE_2$ (prostaglandin $E_2$) and cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$) was reduced by RP3 treatment more than RP1 and RP2. In conclusion, these results indicated that inflammation on Raw 264.7 cells was improved by RP3. Treatment of RP3 could be used to natural medicine for improving inflammatory response. However, further experiment is required to observe how the high temperature extraction at $500^{\circ}C$ for 48 h influences on alteration of active ingredient in Robinia pseudoacacia L., and conducts the inflammation signal pathway on Raw 264.7 cells.

• Food Science and Preservation
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• v.25 no.1
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• pp.98-106
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• 2018

Portulaca oleracea L., a species of Portulacaceae, is ubiquitous. It is a well-known traditional Chinese medicine for removing heat, counteracting toxicity, cooling blood, and maintaining hemostasia; it is also used as antidysentery agent. This study investigated the anti-oxidative and anti-inflammatory activities of water and ethanol extracts from P. oleracea. The total polyphenol content ($21.08{\pm}0.03mg\;GAE/g$) and total flavonoid content ($5.45{\pm}0.76mg\;QE/g$) of the ethanolic extracts were higher than those of the water extracts. The antioxidative activities were determined by evaluating the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity and by the ferric reducing antioxidant potential (FRAP) assay. The ABTS radical scavenging activity of the water extract (75.53%) was higher in those of the water extract (67.03%) at concentration of $1,000{\mu}g/mL$. The DPPH radical scavenging activity and FRAP of the ethanol extract were higher than those of the water extract. We also investigated the anti-inflammatory activity of the P. oleracea extracts in LPS-stimulated Raw 264.7 cells. The production levels of nitric oxide (NO) and reactive oxygen species (ROS) significantly decreased with an increasing concentration of the extract. The expression levels of pro-inflammatory cytokines (tumor necrosis faction (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6) were significantly lower in the ethanol extract than in the LPS alone treatment group. Based on these results, ethanolic extract from P. oleracea could be an effective antioxidant and anti-inflammatory agent.

• Journal of Life Science
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• v.34 no.3
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• pp.160-169
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• 2024

In this study, Abies nephrolepis MAX. was divided into A. nephrolepis MAX. stem (AS) extract and A. nephrolepis MAX. leaf (AL) extract. Their anti-inflammatory abilities and applicability as cosmetic materials were determined. Tests of the cell survival rate measured using RAW 264.7 cells and extracts of AS and AL showed 97.8% and 95.6% cell viability at a 500 ㎍/ml concentration. To determine anti-inflammatory activity, we examined the inhibitory effects on the production of LPS-induced NO in RAW 264.7 cells by Griess assay. The results showed that the AS and AL extracts presented a concentration-dependent inhibition of NO production. The protein expression inhibitory effects of AS and AL extracts were measured by western blot at 25, 50, and 100 ㎍/ml concentrations. β-actin was used as a positive control. The results of western blot of extracts from AS showed that the expression inhibition rate of the iNOS protein was decreased by 50.1% at the 100 ㎍/ml concentration. Additionally, the results of western blot of AL extracts showed that the expression inhibition rate of COX-2 and iNOS protein was decreased by 66% and 8.2% at the 100 ㎍/ml concentration. The mRNA inhibitory effect was measured by RT-PCR at 25, 50, and 100 ㎍/ml concentrations. GAPDH was used as a positive control. Consequently, the iNOS mRNA expression effect by RT-PCR of AS extract demonstrated by RT-PCR decreased by 27.9% at the 100 ㎍/ml concentration, and the iNOS and IL-6 mRNA expression effect of AL extract measured by RT-PCR decreased by 48.6% and 48.7% at the 100 ㎍/ml concentration.