• 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.

• 생명과학회지
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• 제27권5호
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• pp.509-516
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• 2017

들깨(Perilla frutescents (L.) Britton var.) 새싹은 꿀풀과에 속하는 1년생 초본이다. 본 연구의 목적은 들깨 새싹 에탄올 추출물이 사이토카인으로 유도된 췌장 베타 세포 손상에 대한 예방 효과를 평가하기 위함이다. 췌장 소도 주위에 염증 세포 침습으로 의해 분비되는 사이토카인은 1형 당뇨병의 발병원인에 해당된다. 인터루킨-$1{\beta}$ (IL-$1{\beta}$), 인터페론-${\gamma}$ (IFN-${\gamma}$), 종양괴사인자-${\alpha}$ (TNF-${\alpha}$) 등의 사이토카인은 활성산소 형성을 유도한다. 세포 내 활성산소 축적은 췌장 베타 세포 기능장애와 세포사멸을 이끈다. 들깨 새싹 추출물은 항산화 효과를 증가 시켰으며 활성산소 생성을 억제하였다. 사이토카인은 세포생존율을 감소시켰고, iNOS와 COX-2의 발현을 증가시키고 산화질소 생성을 유도하였다. 들깨 새싹 추출물은 사이토카인으로 유도된 세포생존을 농도 의존적으로 예방하였다. 또한, 사이토카인에 의한 산화질소 생성과 iNOS와 COX-2의 단백질 발현 증가를 억제하였다. 더 나아가 들깨 새싹 추출물은 췌장 베타 세포주(RIN-m5F)에서 $I{\kappa}B{\alpha}$ 인산화 억제를 통해서 NF-${\kappa}B$의 활성화를 상당히 감소시켰다. 요약하자면, 본 연구 결과는 들깨 새싹 추출물이 사이토카인으로 유도된 췌장 베타 세포 손상에 대한 보호 효과를 가지고 있다는 것이 확인되었다. 결과적으로 들깨 새싹은 혈당 증가에 의한 산화 스트레스와 염증성 사이토카인에 의한 베타 세포 손상을 완화하여 당뇨에 유익할 것으로 사료된다.

• 동의생리병리학회지
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• 제18권3호
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• pp.880-886
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• 2004

Phellinus igniarius has been clinically used for the treatment of hemorrhoidal fistula, dysmenorrhea and the prevention of cancer in traditional oriental medicine. Recent studies showed that Phellinus igniarius produced anti-cancer, anti-metastasis and immuno-modulatory effects, There is lack of studies regarding the effects of Phellinus igniarius on the immunological activities. The present study was conducted to evaluate the effect of Phellinus igniarius on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in Raw 264,7 cells. After the treatment of Phellinus igniarius water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. COX-2 and iNOS were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that Phellinus igniarius inhibited the production of nitrite and nitrate (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β) and interleukin-6 (IL-6), and the activation of phospholylation of inhibitor κBα (p-IκBα) in Raw 264.7 cells activated with lipopolysaccharide (LPS). These findings suggest that Phellinus igniarius can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

• 생명과학회지
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• 제19권4호
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• pp.479-485
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• 2009

본 연구는 BS 열수추출물의 항염증 활성을 조사하기 위하여 LPS로 자극된 RAW 264.7세포에서 세포가 방출하는 NO, $PGE_2$, COX생성량과 염증성 사이토카인인 $IL-1{\beta}$, IL-6, $TNF-{\alpha}$의 생성량의 변화를 조사하여 항염증성 기능의 유무와 정도를 알아보고자 하였다. 마우스 대식세포인 RAW 264.7에 대한 BS추출물의 농도별(0, 50, 100, $200\;{\mu}g/ml$) 세포독성을 알아보기 위하여 MTT assay를 수행하여 $200\;{\mu}g/ml$ 이하의 농도에서는 세포독성이 낮아 기본적으로 세포의 생존율에 영향을 주지 않는다는 사실을 알 수 있었다. RAW 264.7세포에 BS추출물을 전처리하고 LPS를 18시간 동안 처리한 후 NO의 생성량은 정상세포에 비하여 약 8배 이상 증가되었으며, BS추출물을 $100\;{\mu}g/ml$ 농도 이상으로 처리한 실험군에서는 농도의존적으로 nitrite의 생성량이 유의하게 감소되었다. $PGE_2$의 생성량은 LPS처리에 의하여 정상세포에 비하여 약 14배 이상 증가되었으며 BS추출물을 100 및 $200\;{\mu}g/ml$의 농도로 처리한 실험군에서는 농도의존적으로 유의하게 각각 55.1%, 39.9% 감소되었다. COX의 생성량은 LPS처리에 의하여 정상세포에 비하여 약 5.6배 증가되었으며 $100\;{\mu}g/ml$ 이상의 농도처리군에서는 농도의존적으로 유의하게 감소되었다. LPS처리후 $IL-1{\beta}$, IL-6, $NF-{\alpha}$의 생성량은 정상세포에 비하여 급격하게 증가되었으며, 여러 농도(50, 100, $200\;{\mu}g/ml$)의 BS추출물로 처리한 실험군에서는 각각 아무런 처치를 하지 않은 세포에 비하여 농도의존적인 각각의 생성량이 감소되었다. 특히 $100\;{\mu}g/ml$ 농도 이상의 BS처리군에서는 모두 통계학적 유의성이 인정되었다. 이러한 결과는 BS에서 염증억제 효과가 있음을 in vitro 연구에서 확인할 수 있었고, 향후 유효성분 추출을 통한 항염증 물질의 연구 또는 예방하거나 치료할 수 있는 염증 억제 성분의 분리 및 그 작용기전 연구에 중요한 기초 자료가 될 것이라 사료된다.

• Natural Product Sciences
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• 제16권3호
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• pp.192-197
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• 2010

The seeds of Raphanus sativus L. (RSL) and Phaseolus calcaratus Roxburgh (PHCR), the root of Scutellaria baicalensis (SB), and the flower of Lonicera japonica (LJ) have been traditionally used as herbal medicines for anti-inflammation. Unlike the SB and LJ, little information is available for the scientific bases that show the anti-inflammatory mechanisms of RSL and PHCR. In this study, we prepared boiled water extracts from the medicines and determined their potentials in inhibiting nitric oxide (NO) production, cyclooxygenase-2 (COX-2) expression, and tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6 secretion in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The effects of the medicines on serum IgE levels in ovalbumin (OVA)-administrated mice were also studied. The medicines inhibited production of TNF-$\alpha$ and IL-6, and COX-2 expression in LPSstimulated macrophages. Especially, PHCR water extract showed more potent inhibition on TNF-$\alpha$ production than SB and LJ extracts, but RSL extract did not exert these effects. Similar to the cases of SB and LJ, PHCR extract prevented the phosphorylation of $I{\kappa}B{\alpha}$ and c-Jun, and the activation of NF-${\kappa}B$-DNA binding. Further, oral supplementation of PHCR extract attenuated significantly serum levels of total and OVA-specific IgE in OVAtreated animals. These results suggest a possibility that PHCR water extract can be used for the treatment of inflammatory and allergic diseases.

• 대한본초학회지
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• 제24권3호
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• pp.153-160
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• 2009

Objectives : The purpose of this study was to investigate the anti-inflammatory effects of extract from Trogopterorum Faeces (TF) on the RAW 264.7 cells. Methods : To prove the TF's anti-inflammatory effects, we investigated nitric oxide (NO) production and own cell viability. We examined the cytokine productions on lipopolysacchride (LPS)-induced RAW 264.7 cells and also cellular regulatory mechanisms. Results : TF does not have any cytotoxic effect. TF reduced LPS-induced NO production, interleukin (IL)-1b, IL-6, IL-10 and tumor necrosis factor-a (TNF-a) in RAW 264.7 cells. TF inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-Ba) in the LPS-stimulated RAW 264.7 cells. TF reduced the serum levels of IL-1b, IL-6, TNF-a. The survival rate of LPS-induced endotoxin shock was increased by TF administration. Conclusions : TF down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis for anti-inflammatory properties.

• 대한한방부인과학회지
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• 제18권3호
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• pp.92-109
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• 2005

Purpose : The Purpose of this research was to investigate the effects of Cheongyeolhawlhyeoltanggagyehyeoldeung (CYHHT) on anti-inflammatory effects. Methods : As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators were determined in mouse lung fibroblast cells(mLFC) and RAW264.7 cells. Also, changes in pathological features by drug treatment were investigated in the in vivo edema-induced rats by carrageenin /arachidonic acid or in the colitis-induced mice by DSS treatment. Results : The cytotoxicity of CYHHT on mLFC and RAW264.7 cells was not observed at 100, 50, 10, and $1{\mu}g/ml$ of CYHHT treatments. $IL-1{\beta}$, IL-6 and NOS-IImRNA expression of RAW264.7 cells was inhibited by CYHHT treatments in a dose-dependent manner. CYHHT treatment of RAW264.7 cells inhibited $TNF-{\alpha}$ and COX-2 mRNA expression. CYHHT treatment of RAW264.7 cells significantly inhibited IL-6 and NO production. CYHHT treatment of RAW264.7 cells inhibited ROS production. CYHHT inhibited rat's paw edema induced by carrageenin or arachidonate treatment in all concentrations examined. The body weight and colon length of colitis-induced mice were recovered to a normal level by DSS treatment. Clinical disease levels were significantly improved compared to the control animals. CYHHT treatment of colitis-induced mice significantly increased hematological values such as WBC and RBC counts, Hgb and HCT levels, but decreased PLT values. CYHHT treatment of colitis-induced mice decreased IL-6 and $TNF-{\alpha}$ production significantly CYHHT treatment of colitis-induced mice significantly increased CD3+(T) cell counts. In contrast, CYHHT treatment decreased CD19+ B cell counts and CD3+/CD69+ significantly, and also decreased B/T ratio (%) though not significant. Conclusion : These results indicated that CYHHT could be used for treating diverse female diseases caused by the inflammation.

• 대한한방부인과학회지
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• 제18권4호
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• pp.36-53
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• 2005

Purpose : The Purpose of this research was to investigate the effects of Haingkyunghonghwatang (HKHHT) on anti-inflammatory effects. Methods : As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators were determined in mouse lung fibroblast cells(mLFC) and RAW264.7 cells. Also, changes in pathological features by drug treatment were investigated in the in vivo edema-induced rats by carrageenin/arachidonic acid or in the colitis-induced mice by DSS treatment. Results : The cytotoxicity of HKHHT on mLFC and RAW264.7 cells wasn't observed at 100, 50, 10, and $1{\mu}g/ml$ of The treatments. $IL-1{\beta}$, IL-6 and NOS-II mRNA expression of RAW264.7 cells was inhibited by The treatments in a dose-dependent manner. HKHHT treatment of RAW264.7cells(HtRc) inhibited $TNF-{\alpha}$ and COX-2 mRNA expression. HtRc significantly inhibited IL-6 and NO production. HtRc inhibited ROS production. HKHHT inhibited rat's paw edema induced by carrageenin or arachidonate treatment in all concentrations examined. The body weight and colon length of colitis-induced mice were recovered to a normal level by DSS treatment. Clinical disease levels were significantly improved compared to the control animals. HKHHT treatment of colitis-induced mice(HtCm) significantly increased hematological values such as WBC and RBC counts, Hgb and HCT levels, but decreased PLT values. HtCm decreased IL-6 and $TNF-{\alpha}$ production significantly HtCm significantly increased CD3+(T) cell counts. In contrast, HKHHT treatment decreased CD19+ B cell counts and CD3+/CD69+ significantly, and also decreased B/T ratio (%) though not significant. Conclusion : These results indicated that HKHHT could be used for treating diverse female diseases caused by the inflammation.

• 한국자원식물학회지
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• 제35권5호
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• pp.574-585
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• 2022

순창군에서 생산된 두릅을 40% EtOH을 이용하여 추출한 후 chlorogenic acid 함량, 항산화, 항균, 항염 및 소화효소 활성을 측정하였다. HPLC를 이용하여 chlorogenic acid 함량을 측정한 결과 7.06±0.01 mg/g 함유되어 있었다. DPPH 라디칼 소거활성(SC₅₀)은 4.79±0.05 mg/mL, ABTS 라디칼 소거활성(SC₅₀)은 5.79±0.05 mg/mL, 총 폴리페놀 함량은 170.0±1.8 mgGAE/g, 총 플라보노이드 함량은 105.5±4.1 mgQE/g으로 분석되었다. RAW 264.7 세포, Caco-2 세포에서 세포독성이 나타나지 않았으며, 농도 의존적으로 NO 생성이 억제되었다. RAW 264.7 세포에서 염증성 cytokine인 TNF-α생성은 8.9±0.1 ng/mL, IL-6 생성은 15.2±0.8 ng/mL, IL-1β생성은 30.9±0.9 pg/mL으로 억제되었으며, AEE의 처리 농도가 증가함에 따라 TNF-α, IL-1β, IL-6 생성이 농도 의존적으로 억제되었다. S. typhimurium, L. monocytogenes, H. pylori에 대한 항균활성이 우수하게 나타났으며, 두릅을 EtOH 추출물의 처리농도가 증가함에 따라 효소 활성인 α-amylase와 protease 효소활성도 증가하였다. 순창군에서 생산된 두릅은 chlorogenic acid를 다량 함유하고 있으며, 항산화, 항염, 항균, 소화효소 활성이 우수하게 나타나 향후 건강기능성 소재로 개발할 수 있을 것으로 판단된다.

• 대한한의학회지
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• 제42권4호
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• pp.10-24
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• 2021

Objectives: Yongdamsagan-tang (YST) and Paljung-san (PJS) in traditional medicine and finasteride in modern medicine are used to treat benign prostatic hyperplasia (BPH). In recent, the use of combination herbal remedies with conventional drugs has been increasing. Therefore, we investigated the anti-inflammatory effects of these drugs to treat BPH and the influence of herbal formulas on finasteride metabolism. Methods: The inhibitory effects of the herbal formulas and finasteride on the production of inflammatory mediators and cytokines were determined in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Additionally, the influence of herbal formulas on activities of human drug metabolizing enzymes (DMEs) was assessed using human microsomal enzymes. Results: We observed that YST, PJS and finasteride inhibited the production of nitric oxide (NO), prostaglandin E₂ (PGE₂) and interleukin-6 (IL-6) in RAW 264.7 cells. The half maximal inhibitory concentration (IC₅₀) of YST on PGE₂ production was calculated to be below 25 ㎍/mL. YST inhibited the activity of uridine diphosphate-glucuronosyltransterase (UGT) 1A4 with an IC₅₀ value of 49.35 ㎍/mL. The activities of cytochrome P450 (CYP) 1A2, CYP2B6, CYP2C19, CYP3A4, and UGT1A1 were inhibited by PJS (IC₅₀ < 100 ㎍/mL, each). Although PJS and YST inhibited the activities of CYP3A4 and UGT1A4, respectively, these formulas may not influence the metabolism of finasteride because the IC₅₀ values of herbal formulas on DMEs are too high to affect metabolism. Conclusions: Our results suggest that the combination of finasteride and YST or PJS might not influence their drug metabolism and that the drugs may have synergistic effects against BPH.