• IMMUNE NETWORK
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• 제8권1호
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• pp.7-12
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• 2008

Background: Members belonging to the interferon-lambda (IFN-${\lambda}$) family exert protective action against viral infection; however, the mechanisms of their action have remained elusive. To study IFN-${\lambda}$ biology, such as endocytosis of IFN-${\lambda}$, we produced monoclonal antibodies (Abs) against human IFN-${\lambda}$ and examined their usefulness. Methods: We purified recombinant human IFN-${\lambda}$1 expressed in Escherichia coli by using affinity columns. Then, we generated hybridoma cells by fusing myeloma cells with splenocytes from IFN-${\lambda}$1-immunized mice. For evaluating the neutralizing activity of the monoclonal Abs against IFN-${\lambda}$1, we performed RT-PCR for the MxA transcript. In order to study the binding activity of IFN-${\lambda}$ and the monoclonal Ab complex on HepG2 cells, we labeled the monoclonal Ab with rhodamine and determined the fluorescence intensity. Results: Four hybridoma clones secreting Abs specific to IFN-${\lambda}$1 were generated and designated as HL1, HL2, HL3, and HL4. All the Abs reacted with IFN-${\lambda}$1 in the denatured form as well as in the native form. Abs produced by HL1, HL3, and HL4 did not neutralize the induction of the MxA gene by IFN-${\lambda}$1. We also demonstrated the binding of the HL1 monoclonal anbitody and IFN-${\lambda}$ complex on HepG2 cells. Conclusion: Monoclonal Abs against IFN-${\lambda}$1 were produced. These Abs can be used to study the cellular binding and internalization of IFN-${\lambda}$.

• Archives of Pharmacal Research
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• 제29권10호
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• pp.890-897
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• 2006

In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.

• Biomolecules & Therapeutics
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• 제29권4호
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• pp.410-418
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• 2021

Helicobacter pylori causes chronic gastritis through cag pathogenicity island (cagPAI), vacuolating cytotoxin A (VacA), lipopolysaccharides (LPS), and flagellin as pathogen-related molecular patterns (PAMPs), which, in combination with the pattern recognition receptors (PRRs) of host cells promotes the expression and secretion of inflammation-causing cytokines and activates innate immune responses such as inflammasomes. To identify useful compounds against H. pylori-associated gastric disorders, the effect of chalcone derivatives to activate the nucleotide-binding oligomerization domain (NOD)-like receptor family, pyrin domain-containing 3 (NLRP3) inflammasome was examined in an H. pylori-infected human monocytic THP-1 cell line in this study. Among the five synthetic structurally-related chalcone derivatives examined, 2'-hydroxy-4',6'-dimethoxychalcone (8) and 2'-hydroxy-3,4,5-trimethoxychalcone (12) strongly blocked the NLRP3 inflammasome in H. pylori-infected THP-1 cells. At 10 μM, these compounds inhibited the production of active IL-1β, IL-18, and caspase-1, and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) oligomerization, but did not affect the expression levels of NLRP3, ASC, and pro-caspase-1. The interruption of NLRP3 inflammasome activation by these compounds was found to be mediated via the inhibition of the interleukin-1 receptor-associated kinase 4 (IRAK4)/IκBα/NF-κB signaling pathway. These compounds also inhibited caspase-4 production associated with non-canonical NLRP3 inflammasome activation. These results show for the first time that certain chalcones could interrupt the activation of the NLRP3 inflammasome in H. pylori-infected THP-1 cells. Therefore, these chalcones may be helpful in alleviating H. pylori-related inflammatory disorders including chronic gastritis.

• 대한본초학회지
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• 제29권6호
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• pp.117-123
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• 2014

Objectives : Sinhyowoldo-san (SHWDS) is said to be a traditional medicine used for shigellosis, abdominal pain, diarrhea. But mechanism of SHWDS mediated-modulation of immune function is not sufficiently understood. To ascertain the molecular mechanisms of SHWDS 70% EtOH extract on pharmacological and biochemical actions in inflammation, we researched the effect of pro-inflammatory mediators in phorbol-12-myristate-13-acetate (PMA)+ A23187-activated human mast cell line (HMC-1). Methods : In the present research, cell viability was measured by MTS assay. pro-inflammatory cytokine production was measured by performing enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western blot analysis to analyze the activation of mitogen-activated protein kinases (MAPKs), nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). The investigation focused on whether SHWDS inhibited the expressions of interleukin-6 (IL-6), interleukin-8 (IL-8), MAPKs and $NF-{\kappa}B$ in PMA+A23187-activated HMC-1 cells. Results : SHWDS has no cytotoxicity at measured concentration (50, 100, and $250{\mu}g/ml$). SHWDS ($250{\mu}g/ml$) inhibits pro-inflammatory cytokine expression in PMA+ A23187-activated HMC-1 cells. Moreover, SHWDS inhibited cyclooxygenase (COX)-2 expression. In activated HMC-1 cells, SHWDS suppressed phosphorylation of extracellular signal-regulated kinase (ERK 1/2) and c-jun N-terminal Kinase (JNK 1/2). Then, SHWDS suppressed activation of nuclear factor $NF-{\kappa}B$ in nuclear, degradation of IkB ${\alpha}$ in cytoplasm. Conclusions : We propose that SHWDS has an anti-inflammatory therapeutic potential, which may result from inhibition of ERK 1/2, JNK 1/2 phosphorylation and $NF-{\kappa}B$ activation, thereby decreasing the expression of pro-inflammatory genes.

• 한국식품영양학회지
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• 제29권3호
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• pp.431-437
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• 2016

본 연구는 전통약재로 주로 사용되는 구절초에서 분리한 다당류(CZPS)가 선천 및 적응면역에서 중추적인 역할을 수행하는 대식세포의 활성화에 미치는 영향에 관하여 관찰하였다. 구절초에서 분리한 다당류를 마우스 유래 대식세포주인 RAW 264.7 cell에 처리하였을 때, iNOS의 발현을 조절하여 NO의 생성이 증가되었으며, 또한 면역활성 물질인 TNF-${\alpha}$, IL-$1{\beta}$ 및 IL-6 등의 cytokine의 분비능이 증가되었으며, 이러한 면역활성 매개자인 NO 및 cytokine의 증가의 원인에 관한 정확한 면역세포내 신호전달에 관하여 알아본 결과, CZPS 처리는 MAPKs(ERK, p38)의 인산화를 촉진시키며, 이로 인한 전사인자인 NF-${\kappa}B$의 인산화를 증가시켜, 대식세포의 활성화를 유도시키는 것으로 관찰되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.749-756
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• 2014

The present study was carried out to investigate the effects of dietary antioxidants on pro-inflammatory cytokines, heat shock protein (HSP) and antioxidant status in broiler chicks under summer conditions. A total of 162, 3-d-old broiler chicks were randomly assigned to a basal diet (CON) and the basal diet supplemented with vitamin C (200 mg/kg diet, VCD) or vitamin E (100 mg/kg, VED) until 35 day of age. All birds were exposed to summer diurnal heat stress at average daily fluctuations of temperature between $32^{\circ}C$ to $34^{\circ}C$ at day to $27^{\circ}C$ to $29^{\circ}C$ at night for the entire feeding periods. There was no significant difference in body weight, feed to gain ratio and the relative organ weight except the thymus in response to dietary vitamin C or E supplementation. However, the mRNA expression of interleukin (IL)-$1{\beta}$, IL-6, interferon (IFN)-${\gamma}$, Toll like receptor (TLR)-4 and HSP70 in the liver of birds fed diet containing vitamin C significantly (p<0.05) decreased compared with those in birds fed basal diet. Dietary vitamin E also showed a significant (p<0.05) decrease in the mRNA expression of IL-6 and HSP70 compared with a basal diet. Total antioxidant status (TAS) in serum of birds fed vitamin C supplemented diet was significantly (p<0.05) higher with than that in birds a basal diet. Lipid peroxidation in serum and liver resulted in a significant (p<0.05) decrease in response to dietary vitamin C or E supplementation. In conclusion, dietary supplementation with antioxidant vitamins, especially vitamin C resulted in a significant decrease in the mRNA expression of pro-inflammatory cytokines and HSP70, and higher antioxidant parameters than that of birds on the basal diet under summer conditions.

• 대한본초학회지
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• 제38권4호
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• pp.21-29
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• 2023

Background : The purpose of this study was to investigate the anti-inflammatory properties of stems and leaves of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. (ES) from Gangwon-do. Methods and Results : Stems and leaves of ES were collected from two areas in Gangwon-do: Cheorwon-gun and Samcheok-si. Samples were extracted with water by using the pressurized liquid extraction method and with 70% prethanol A by using the heat reflux extraction method. The anti-inflammatory effects of ES were evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT), lactate dehydrogenase(LDH) assay, nitric oxide(NO) assay, enzyme-linked immunosorbent assay(ELISA), and Western blot analysis in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). 1) Results showed that ES leaf extractions were not cytotoxic at a concentration of up to 30 ㎍/㎖. The leaves of 70% prethanol A extractions of ES(30 ㎍/㎖) inhibited NO, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) production and decreased the protein level of cyclooxygenase 2(COX-2). There was no significant change in the protein level of inducible nitric oxide synthase(iNOS). The stem extractions of ES did not exhibit anti-inflammatory effects. Conclusions : In this study, the leaves of 70% prethanol A extractions of ES demonstrated anti-inflammatory effect on RAW 264.7 macrophages. The 70% prethanol A extractions have a relatively higher anti-inflammatory effect on RAW 264.7 macrophages than water extractions.

• 대한본초학회지
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• 제29권4호
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• pp.83-89
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• 2014

Objectives : The purpose of this study was to investigate effects of Houttuyniae Herba water extract (HC) on calcium release and production of various inflammatory mediators such as nitric oxide (NO), interferon-inducible protein (IP)-10, platelet derived growth factor (PDGF)-BB, keratinocyte-derived chemokine (KC), vascular endothelial growth factor (VEGF), interleukin (IL)-4, and IL-5 in lipopolysaccharide (LPS)-induced RAW 264.7 mouse macrophages. Methods : NO production was measured by Griess reagent assay. Intracellular calcium level was measured with Fluo-4 assay. Levels of cytokines were measured by High-throughput multiplex bead array cytokine assay based on xMAP (multi-analyte profiling beads) technology. Results : HC significantly decreased NO production for 24 hrs incubation at the concentrations of 10, 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC significantly decreased production of IP-10, KC, VEGF, and PDGF-BB for 24 hrs incubation at the concentrations of 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). HC also significantly decreased intracellular calcium release for 24 hrs incubation at the concentrations of 25, 50, 100, and $200{\mu}/mL$ in LPS-induced RAW 264.7 (P < 0.05). But HC did not show any significant effect on production of IL-4 and IL-5 in LPS-induced RAW 264.7. Conclusions : The results suggested that HC has anti-inflammatory property related with its inhibition on the production of NO, IP-10, KC, VEGF, and PDGF-BB in LPS-induced macrophages via calcium pathway.

• 한국식품영양학회지
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• 제29권6호
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• pp.998-1007
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• 2016

본 연구에서는 상추 추출물이 혈관내피세포의 염증반응에 미치는 영향 및 지질대사 개선 효과를 확인하기 위해 HUVEC을 이용한 in vitro 실험 및 고콜레스테롤혈증 모델을 이용한 동물실험을 수행하였다. 상추 추출물은 HUVEC에서 TNF-${\alpha}$에 의해 증가되는 세포접착단백질 ICAM-1, VCAM-1의 발현을 억제하였고 TNF-${\alpha}$에 의해 촉진되는 HUVEC와 단핵구 세포의 부착을 감소시켰다. 또한 상추 추출물은 TNF-${\alpha}$에 의해 증가되는 염증성 사이토카인 IL-6, IL-8 및 동맥경화 유발 케모카인 MCP-1의 생성을 농도 의존적으로 억제하였다. 고콜레스테롤혈증 동물모델에서 고콜레스테롤 식이군 및 상추 추출물이 첨가된 고콜레스테롤 식이군의 체중증가량 및 식이섭취량은 유의적인 차이를 보이지 않았다. 고콜레스테롤 식이군의 혈중 총 콜레스테롤 함량은 정상 식이군에 비해 크게 증가하였고 상추추출물이 첨가된 고콜레스테롤 식이군의 콜레스테롤 함량은 유의적으로 감소하였다. 고콜레스테롤 식이군의 혈중 중성지방 함량은 정상 식이군에 비해 약간 감소하였지만 유의적인 차이는 보이지 않았다. 상추 추출물이 첨가된 고콜레스테롤 식이군의 경우 혈중 중성지방 함량은 고콜레스테롤 식이군과 비교하였을 때 유의적인 차이를 보이지 않았다. 고콜레스테롤 식이군의 HDL-콜레스테롤 함량은 정상 식이군에 비해 감소하였고 상추 추출물이 첨가된 경우 HDL-콜레스테롤 함량은 고콜레스테롤 식이군에 비해 유의적으로 증가하였다. LDL-콜레스테롤 함량은 고콜레스테롤 식이군에서 크게 증가하였고 상추 추출물이 첨가된 경우 감소하였다. 혈청의 동맥경화지수 및 심혈관지수는 고콜레스테롤 식이군에서 정상 식이군에 비해 유의적으로 증가하였고 상추 추출물이 첨가된 고콜레스테롤 식이군에서 유의한 감소를 나타내었다. 이상의 연구결과는 상추 추출물이 혈관내피세포의 염증반응을 억제하여 혈관협착을 방지할 수 있고 혈중 지질대사의 개선 효과를 보여 혈액순환개선 및 동맥경화, 심혈관질환의 예방과 치료에 효과가 있다는 것을 의미한다.

• Tuberculosis and Respiratory Diseases
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• 제48권6호
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• pp.922-931
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• 2000

배경 및 목적 : 기관지천식의 중요한 특징적인 병인인 기도의 만성염증 상태와 증상의 변화를 판단하는 객관적인 지표로서 여러 가지 방법이 시도되고 있는데 최근에는 비침습적이고 비교적 간단한 유도객담 검사로 침습적인 기관지 조직 검사나 기관지폐포세척술을 대신하려는 경향이 있다. 저자들은 기관지천식 환자의 유도객담에서 호산구, eosinophil cationic protein(ECP), interleukin(IL)-3, IL-5, granulocyte-macrophage colony-stimulating factor(GM-CSF), 그리고, nitric oxide(NO)의 유도체를 측정하고, 객담내 NO 및 호산구와 관계가 있다고 알려진 cytokine들의 농도가 증상 및 기도의 염증상태를 반영하는 지표로서 이용될 수 있는지를 알아보고자 하였다. 방법 : 기관지천식 환자 30명을 대상으로 2주간 경구용 프레드니솔론 30mg을 투여하고, 약제 투여 전후로 1초간 노력성 호기량($FEV_1$과 혈중 총 호산구수, 그리고 유도객담 검사를 시행하여 객담내 호산구 비율과 ECP, IL-3, IL-5, GM-CSF, 그리고, NO 유도체를 측정하였다. 결과 : 대상환자중 남자는 13a여(43.3%), 여자는 17명(56.7%)이었고 연령분포는 19세부터 64세까지였으며 평균연령은 41.8세였다. 30명중 2명은 두 번째 검사시 객담이 채취되지 않았고, 3명은 두 번째 방문을 하지 않았다. 프레드니솔론 투여후 평균 1초당 노력성 호기량(% of predicted value)은 78.1$\pm$27.2%에서 90.3$\pm$18.3%로 개선되었고(P<0.001), 유도객담내 평균 호산구비율은 56.1$\pm$27.2%에서 29.6$\pm$21.3%로 (P<0.001), 객담내 평균 ECP 농도는 $134.5\pm68.1{\mu}\;g/L$에서 $41.5\pm42.4\;{\mu}g/L$로 유의하게 감소하였고(P<0.001), 평균 혈중 총 호산구수는 425.7$\pm$265.9에서 287.7$\pm$294.7로 감소하는 경향을 보였지만 통계적으로 유의하지 않았다. 유도객담내 NO 유도체의 평균 농도는 프레드니솔론 투여 전호로 유의한 차이가 없었으며($70.4\pm44.6\;{\mu}mol/L$ vs $91.5\pm48.3\;{\mu}mol/L$) 평균 eotaxin 농도는 27.7$\pm$12.8 pg/ml에서 21.7$\pm$8.7 pg/ml로 감소하는 경향을 보였다. IL-3, IL-5, CSF는 대부분의 경우에서 측정되지 않았다. 치료전의 1초당 노력성 호기량과 객담내 ECP의 농도 사이에 의미있는 역상관관계가 있었고(r=0.369, P<0.05). 결론 : 유도객담내에서 측정 가능한 여러 인자중 eotaxin 및 ECP가 치료에 따른 기도염증 상태의 변화를 반영하는 지표로 이용될수 있을 것으로 생각된다. 그 외 유도객담내 호산구비율 측정도 치료에 따른 증상이나 염증의 변화를 손쉽게 관찰할수 있는 간단한 방법으로 생각되지만 유도객담에서 Griess reagent를 이용한 NO 유도체의 측정은 기관지천식의 증상이나 염증상태의 변화를 반영하는 지표로서 유용하지 않을 것으로 생각된다.