• 생명과학회지
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• 제32권2호
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• pp.125-134
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• 2022

본 연구는 노니(Morinda citrifolia)에 함유된 주요 생리활성물질이 RAW 264.7 세포에서 JAK/STAT 신호전달 경로를 통하여 항염증 작용에 관여하는 것을 조사하였다. 건조된 M. citrifolia 열매의 MeOH 추출에 의한 유기용매의 순차 분획에서 얻은 EtOAc 분획물(Mc-EtOAc)에서 가장 높은 항산화 활성을 확인하였고, H₂O₂로 유도된 RAW 264.7 세포의 산화적 스트레스에 대한 세포보호 효과는 처리 농도의존적으로 세포독성을 차단하였다. LPS 처리로 71.6%의 RAW 264.7 세포 생존율에서 240 ㎍/ml의 Mc-EtOAc를 전처리한 군에서 84.5%의 세포 생존율 증가를 보였다. LPS로 유도된 RAW 264.7 세포의 NO 생성 저해활성은 240 ㎍/ml의 Mc-EtOAc에서 양성 대조군의 절반 수준으로 NO의 생성양을 감소시켰다. Mc-EtOAc 처리로 iNOS의 발현량은 농도의존적으로 유의하게 감소하였고, COX-2의 발현은 LPS 유도로 3배 정도로 증가되었으나, 120, 240 ㎍/ml의 Mc-EtOAc를 전처리시 농도의존적으로 유의하게 감소시켰다. 또한 pro-inflammatory cytokine IL-1β와 TNF-α의 mRNA 발현도 억제하였다. 이러한 Mc-EtOAc의 항염증 작용이 상위 신호전달 경로인 JAK/STAT 신호전달 경로 통해 일어나지를 조사한 결과, Mc-EtOAc의 전처리로 LPS로 유도된 RAW 264.7 세포에서 pJAK1과 pSTAT3의 인산화 정도가 유의성 있게 감소하였다. 따라서 RAW 264.7 세포에서 Mc-EtOAc의 항염증 효과는 JAK1/STAT3 신호전달 경로를 통해 염증반응을 억제하는 것으로 사료된다.

• 한국미생물·생명공학회지
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• 제45권2호
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• pp.101-109
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• 2017

본 실험에서는 참치 심장 70% ethanol 추출물을 dichloromethane으로 분획한 후 항염증 효과를 확인하기 위해 RAW 264.7 세포에 LPS로 염증을 유도시켜 염증 매개성 물질인 NO와 pro-inflammatory cytokine의 분비량의 변화를 확인하였다. 그 결과, 참치 심장 dichloromethane 분획물을 처리하였을 때, 농도 의존적으로 NO의 생성량을 감소시키는 것을 확인하였으며, 특히, $100{\mu}g/ml$에서 가장 높은 억제효과를 나타내었다. 따라서 dichloromethane 분획물의 억제 활성이 세포사멸에 의한 감소인지 알아보기 위해서 MTT assay를 하였을 때, 세포 생존율이 dichloromethane 분획물을 PBS 처리군과 비교하였을 때 유의적인 차이가 나타나지 않음을 확인하였고, 이를 통해 dichloromethane 분획물이 NO 및 전염증성 cytokine의 분비를 효과적으로 억제할 수 있는 물질임을 확인할 수 있었다. Dichloromethane 분획물을 처리하였을 때, 염증 관련 단백질 발현 정도를 western blot을 통해 확인한 결과, LPS에 의해 발현이 증가된 $NF-{\kappa}B$, iNOS 및 COX-2는 분획물을 처리함으로써 농도 의존적으로 감소되는 것을 확인할 수 있었다. 또한 dichloromethane 분획물의 처리가 인산화된 MAPKs의 발현을 저해함을 확인하여 참치 심장 dichloromethane 분획물이 $NF-{\kappa}B$와 MAPKs의 발현을 억제시킴으로써 NO 및 pro-inflammatory cytokine의 분비량을 감소시킴을 확인할 수 있었다. 동물 모델에서는, dichloromethane 분획물을 처리하였을 때 croton oil에 의한 귀 부종이 농도 의존적으로 감소함을 확인하였고, 특히, $250mg/kg{\cdot}body\;weight$ 농도로 투여시 시판 항염증제인 predinisolone을 $50mg/kg{\cdot}body\;weight$ 농도로 투여한 그룹과 유사한 효과를 나타내었다. 조직학적 변화를 확인한 결과에서는, 진피와 경피의 두께가 감소하였으며 진피내 mast cell 침윤이 감소되는 것을 확인할 수 있었다. 따라서 참치심장 dichloromethane 분획물이 효과적인 염증 예방 및 부종 완화를 위한 치료제로서 활용 가능성을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권12호
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• pp.1753-1759
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• 2008

Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

• 생명과학회지
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• 제26권6호
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• pp.640-645
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• 2016

본 연구에서는 녹차 유래 polyphenol 중의 하나인 epigallocatechine gallate (EGCG)를 이용한 신경염증 억제 효과를 확인하였다. LPS로 유도된 미세아교세포의 활성화로 분비되는 nitric oxide (NO)와 pro-inflammatory cytokine을 포함하여 iNOS, TNF-a와 IL-1b 유전자의 발현과 LPS 수용체인 TLR-4의 활성에 미치는 EGCG의 억제 효능을 확인하였다. Latex beads를 이용한 phagocytotic activity를 확인한 결과 LPS로 유도된 미세아교세포 활성에 의한 식균활성이 EGCG에 의해 억제되는 것을 볼 수 있었다. 뿐만 아니라, BV-2 미세아교세포 조건배지를 이용하여 도파민성 신경세포 SN4741의 세포 사멸확인에서도 EGCG에 의한 보호 효과를 확인하였다. 본 연구 결과는 녹차 유래 polyphenol인 EGCG의 신경염증 반응억제효능과 신경퇴행성 질환 제어 가능성을 확인하였다. 본 연구의 결과는 녹차 유래 polyphenol인 EGCG의 신경염증 반응과 그로 인한 신경 퇴행성 질환 제어 가능성을 제시하였다.

• 대한본초학회지
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• 제22권4호
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• pp.117-125
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• 2007

Objectives : Berberine, a main alkaloid component of Coptidis rhizoma, has an antimicrobial and anti-tumor activities and antiinflammatory effect. In the present study, we investigated effect of berberine on the production of inflammatory mediators such as nitric oxide(NO), prostaglandin E2(PGE2), TNF-${\alpha}$ and IL-1${\beta}$ in LPS-stimulated BV2 microglial cells, Methods : BV2 cells were pre-treated with berberine and then stimulated with LPS. The cytotoxicity of berberine was determined by MTT assay. The NO production was measured by Griess assay. The mRNA expression and protein levels of inducible nirtic oxide synthase(iNOS) were determined by RT-PCR and Western blot. The production of PGE2 and cytokines was measured by ELISA. Results : Berberine inhibited the production of NO, PGE2 and pro inflammatory cytokines, TNF-${\alpha}$ and IL-1${\beta}$ in a dose dependent manner in LPS-stimulated BV2 cells. In addition, berebrine greatly suppressed the mRNA expression and protein levels of iNOS and inflammatory cytokines induced by LPS stimulation. These results indicate that the post-transcriptional regulatory mechanism of iNOS and/or inflammatory cytokine gene expression by berberine is involved in its anti-inflammatory effects, respectively. Conclusion : The present study suggests that berberine can be useful as a potential anti-inflammatory agent for treatment of various neurodegenerative diseases such as Alsheimer's disease, Parkinson's disease and stroke.

• 대한본초학회지
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• 제31권1호
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• pp.69-75
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• 2016

Objectives : The aim of this study is to investigate the various effects of individual or combined extract of GamiSaengmaeksan (GSS) on cell viability, anti-inflammatory and antioxidant activityMethods : In order to evaluate cytotoxicity, MTT assay was performed. We investigated the levels of proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 and interleukin (IL)-1β, and nitric oxide(NO) in LPS-induced RAW 264.7 cells to check the effects on anti-inflammatory activity. The level of NO production in RAW 264.7 cells was measured by using Griess reagent. The levels of cytokines and ROS were measured by Luminex and Flow cytometry, respectively.Results : At concentration of 200 ㎍/㎖ GSS, cytotoxicity was observed in RAW 264.7 cells. However, at concentration less than 100 ㎍/㎖ of both combine and individual GSS, cytotoxicity was not observed in Raw 264.7 cells. However, the level of ROS in RAW 264.7 cells were decreased at both extract of 100 ㎍/㎖ GSS. Also, the level of NO in RAW 264.7 cells were decreased from extraction of concentration of 100 ug/ml in GSS and individual-extraction of Liriopis Tuber, White Ginseng and Glycyrrhizae Radix. In addition, productions of pro-inflammatory cytokines (TNF-α) in LPS-induced RAW 264.7 cells were decreased from extraction of concentration of 10 and 100 (㎍/㎖) in GSS and individual-extraction of Liriopis Tuber.Conclusions : It is concluded that combined extract of GSS appears to be more effective in anti-oxidation and anti-inflammatory effect than those in individual-extraction of GSS. These results may be developed as a raw material for new therapeutics to ease the symptoms related with inflammatory and oxidative stress.

• BMB Reports
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• 제51권6호
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• pp.308-313
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• 2018

Small-molecule inhibitors are widely used to treat a variety of inflammatory diseases. In this study, we found a novel anti-inflammatory compound, 1-[(2R,4S)-2-methyl-4-(phenylamino)-1,2,3,4-tetrahydroquinolin-1-yl]prop-2-en-1-one (MPQP). It showed strong anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. These effects were exerted through the inhibition of the production of NO and pro-inflammatory cytokines, such as interleukin (IL)-6, $IL-1{\beta}$, and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$). Furthermore, MPQP decreased the expression levels of inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2). Additionally, it mediated the inhibition of the phosphorylation of p38, c-Jun N-terminal kinase (JNK), the inhibitor of ${\kappa}B{\alpha}$ ($I{\kappa}B{\alpha}$), and their upstream kinases, $I{\kappa}B$ kinase (IKK) ${\alpha}/{\beta}$, mitogen-activated protein kinase kinase (MKK) 3/6, and MKK4. Furthermore, the expression of IL-1 receptor-associated kinase 1 (IRAK1) that regulates $NF-{\kappa}B$, p38, and the JNK signaling pathways, was also increased by MPQP. These results indicate that MPQP regulates the IRAK1-mediated inflammatory signaling pathways by targeting IRAK1 or its upstream factors.

• 대한본초학회지
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• 제23권3호
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• pp.93-102
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• 2008

Objectives : Forsythiae Fructus (Forsythia koreana Nakai) has been used anti-inflammatory, diuretics, antidote, and antibacterials in traditional herbal medicine. The present study is focused on the inhibitory effect of Forsythiae Fructus ethanol extract (FF-E) on the production of inflammatory mediators such as NO, iNOS and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$ and IL-6) in LPS-stimulated BV-2 cells, a mouse microglial cell line, and investigated the scavenging activity of FF-E. Methods : BV-2 cells were pre-incubated with FF-E for 30 min and then stimulated with LPS (1 ${\mu}g/m{\ell}$) at indicated times. Cell toxicity of GCF was determined by MTT assay. The levels of NO, PGE2 and cytokines were measured by Griess assay and ELISA. The mRNA and protein expressions of iNOS and cytokines were determined by RT-PCR and Western blotting. Free radical scavenging activity of GCF was determined by DPPH assay in tube test. Results : FF-E significantly inhibited the excessive production of NO, $PGE_2$, $TNF-{\alpha}$, and $IL-1{\beta}$ in LPS-stimulated BV-2 cells. In addition, FF-E attenuated the mRNA and protein expressions of iNOS, and proinflammatory cytokines. FF-E also significantly scavenged the DPPH free radicals in a dose-dependent manner. Conclusions : These results indicate that FF-E exhibits anti-inflammatory property by suppressing the transcription of inflammatory mediator genes, suggesting the anti-inflammatory property of FF-E may make it useful as a therapeutic agent for the treatment of human neurodegenerative diseases.

• 동의생리병리학회지
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• 제35권6호
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• pp.235-241
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• 2021

Sogunjung-tang (SGJT) is traditional herbal prescriptions used to treat abdominal pain. In this study, we evaluated the effect on inflammation and gastritis through SGJT formulation development. SGJT composition herbal medicine was boiled in water at 95~100℃ for 3 hours and then concentrated. Sogunjung-tang mix soft extract (STM) was prepared using pharmaceutical excipients such as purified water, sodium benzoate, β-cyclodextrin and CMC-Na. Anti-inflammatory experiment was conducted using STM and lipopolysaccharide (LPS) in RAW 264.7 cells. Cell survival was measured by MTT method. Nitric oxide (NO) was measured using griess reagents, and pro inflammatory cytokines were measured by enzyme-linked immunosorbent assay and RT-PCR. Also, we verified STM validity in acute gastritis using the ICR mouse. STM was administered oral for three days. And 150 mM HCl in 60% ethanol was oral administered 0.5 mL one hour after the last drug administration. Mice were sacrificed 1 hour after 150 mM HCl in 60% ethanol administration. The gastric mucosa was visually observed. STM were not toxic in RAW 264.7 cells. Treatment of STM inhibited the production of NO and inflammatory cytokine at the protein and mRNA levels. Also, in the acute gastritis model with the mouse, the treatment of STM improved gastric mucosal bleeding and edema. In summary, it was confirmed that the treatment of STM exerts anti-inflammatory and anti-gastritis effects. Therefore, we suggest that STM may provide a preclinical evidence for the prevention and treatment of inflammatory and gastritis diseases.

• 대한화장품학회지
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• 제46권2호
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• pp.195-204
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• 2020

본 연구에서는 피부각질형성세포에서 동의보감에 기록된 14 종의 충부약재에 대한 염증성 사이토카인 생성에 미치는 영향을 조사하였다. 이를 확인하기 위하여 멀티플렉스 사이토카인 분석으로 17 종에 대한 사이토카인을 스크리닝 하였다. 충부약재 14 종 중, 제조 및 전갈 추출물은 IL-5 생성을 억제하였고, 제조, 상표초 및 수질 추출물은 IL-6 생성을, 제조, 상표초, 지룡, 수질 및 문합 추출물은 IL-8 생성에 억제 효과를 보였다. 합개, 잠사, 선퇴 및 문합 추출물은 IL-13 생성을 억제하였으며, 귀판, 잠사 및 지룡 추출물은 MIP-1β 생성에 유의적인 효과를 확인하였다. 이러한 충부약재는 피부염증 억제를 위한 항염증제 활용 가능성에 의미를 둘 수 있지만, 그 가치를 증명하기 위해서는 피부개선을 위한 기전연구를 포함하여 인체피부모사판 모델에서 추가 실험을 통한 검증이 선행 되어야 한다.