• Biomolecules & Therapeutics
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• 제22권1호
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• pp.10-16
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• 2014

Derivatives of caffeic acid have been reported to possess diverse pharmacological properties such as anti-inflammatory, anti-tumor, and neuroprotective effects. However, the biological activity of methyl p-hydroxycinnamate, an ester derivative of caffeic acid, has not been clearly demonstrated. This study aimed to elucidate the anti-inflammatory mechanism of methyl p-hydroxycinnamate in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Methyl p-hydroxycinnamate significantly inhibited LPS-induced excessive production of pro-inflammatory mediators such as nitric oxide (NO) and $PGE_2$ and the protein expression of iNOS and COX-2. Methyl p-hydroxycinnamate also suppressed LPS-induced overproduction of pro-inflammatory cytokines such as IL-$1{\beta}$ and TNF-${\alpha}$. In addition, methyl p-hydroxycinnamate significantly suppressed LPS-induced degradation of $I{\kappa}B$, which retains NF-${\kappa}B$ in the cytoplasm, consequently inhibiting the transcription of pro-inflammatory genes by NF-${\kappa}B$ in the nucleus. Methyl p-hydroxycinnamate exhibited significantly increased Akt phosphorylation in a concentration-dependent manner. Furthermore, inhibition of Akt signaling pathway with wortmaninn abolished methyl p-hydroxycinnamate-induced Akt phosphorylation. Taken together, the present study clearly demonstrates that methyl p-hydroxycinnamate exhibits anti-inflammatory activity through the activation of Akt signaling pathway in LPS-stimulated RAW264.7 macrophage cells.

• 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.

• Journal of Applied Biological Chemistry
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• 제63권1호
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• pp.103-110
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• 2020

Objective: The objective of our study was to investigate anti-cancer effects of Danggui Liuhuang Decoction extract bioconverted by protease liquid coenzyme of Aspergillus kawachii (DLD-BE), compared to a non-bioconverted DLD extract (DLD-E) and determine the underlying mechanisms. Methods: DLD-E and DLD-BE were evaluated for their ability to modulate these signaling pathways and suppress the proliferation of human colorectal cancer (CRC) cells, HCT-116, LoVo, and HT-29. The anti-cancer effects of DLD-E and DLD-BE were measured by using proliferation and migration assays, cell cycle analysis, Western blots, and real-time PCR. Results: In this study, treatment with DLD-E and DLD-BE at concentrations of 25-100 ㎍/mL inhibited proliferation and migration in human CRC cells. DLD-BE induced apoptotic cell death and decreased COX-2 expression in HT-29 cells. The mechanisms of action included modulation of the AKT and extracellular-signal-regulated kinase signaling cascades along with inhibition of COX-2 expression. The results demonstrate novel anti-cancer mechanisms of DLD-BE against the growth of human CRC cells. Thus, we propose that DLD-BE can be developed as a more potent supplement to inhibit colorectal tumor growth and intestinal inflammation than DLD-E.

• 한방안이비인후피부과학회지
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• 제23권1호
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• pp.23-43
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• 2010

Objective : This study was performed to assess the effects of Aloe and Violae herba extracts on skin disease and skin beauty. Methods : Anti-oxidant effects were measured by the scavenging for DPPH radical, xanthine oxidase activity. Anti-inflammatory effects were examined by relations in NO synthesis, IL-$1{\beta}$, IL-6, TNF-$\alpha$, NF-kB, COX-2, MAP kinase. The skin wrinkle formation effects were measured by collagenase and elastase activities. The whitening effects were examined by tyrosinase activities, melanin synthesis in MNT-1 cell. Results : 1. In an anti-oxidant test, Aloe and Violae herba extracts showed high radical scavenging activity. 2. In an anti-inflammatory test, Aloe and Violae herba extracts strongly inhibited the lipopolysaccharide(LPS)-induced nitric oxide(NO) release from the RAW 246.7 macrophage cells. Aloe and Violae herba extracts also inhibited the LPS-induced IL-$1{\beta}$ and COX-2 expressions. The inhibitory effects of Aloe and Violae herba extracts on macrophage activation were via the inhibition of NF-kB, evidenced by transient transfection assay. Furthermore, Aloe and Violae herba extracts weakly inhibited the activation of Jun-N-terminal kinase(JNK) but they did not have any effects on p38 MAP kinase in RAW 264.7 cells. 3. In the skin wrinkle formation assay, Aloe extract strongly inhibited collagenase and elastase, whose activity are tightly related with the wrinkle formation. 4. In the skin whitening assay, Aloe and Viloae herba extracts weakly inhibited tyrosinase activity, however, it was not statistically significant. Besides they did not have any effects on melanin synthesis, indicating that they could not be applicable for skin whitening. Conclusion : This study show that Aloe and Violae herba extracts may play a significant role in skin disease and skin beauty.

• 농업과학연구
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• 제46권3호
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• pp.653-660
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• 2019

Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

• 대한한의학방제학회지
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• 제27권1호
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• pp.7-16
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• 2019

Objectives : Hemistepta lyrata Bunge (Bunge) has been used for treating wound, hemorrhage, fever in Korean traditional medicine. Present study investigated anti-inflammatory effect of H. lyrata chloroform extract (HLE) and its molecular mechanism involved. Methods : To assess anti-inflammatory effect of HLE, production of nitric oxide (NO) and expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines were measured on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Moreover, nuclear factor-${\kappa}B$ (NF-${\kappa}B$) signaling pathway was examined to elucidate its molecular mechanism. Results : Pretreatment of HLE inhibited NO production in a concentration dependent manner. HLE also decreased expression of iNOS and COX-2, and alleviated expressions of pro-inflammatory cytokines in LPS-stimulated RAW 264.7 cells. Moreover, HLE pretreatment inhibited phosphorylation of inhibitory-${\kappa}B{\alpha}$ and p65. Conclusions : These results suggest that HLE exhibits anti-inflammatory effect via inhibition of NF-${\kappa}B$.

• Journal of Applied Biological Chemistry
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• 제63권4호
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• pp.421-428
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• 2020

본 연구는 황기와 지치 복합물인 ALM16이 lipopolysaccharide 처리에 의해 자극된 RAW264.7 대식세포의 염증반응에 미치는 영향에 대하여 조사하였다. ALM16은 RAW264.7 대식세포에 대하여 최대 200 ㎍/mL의 농도까지 독성은 보이지 않았다. 항염증 활성을 검정하기 위해 nitric oxide (NO), prostaglandin E2 (PGE2) 및 pro-inflammatory cytokines 생성량을 측정한 결과, ALM16은 각각의 생성량을 농도의존적으로 감소시켰다. 또한 ALM16은 NO와 PGE2 생성에 관여하는 inducible nitric oxide synthase (iNOS)와 cyclooxygenase-2 (COX-2)의 단백질 발현을 억제하였다. 한편, 항염증 활성 조절 기전을 확인하기 위하여 NK-κB의 핵으로의 이동과 DNA-binding activity 및 MAPK 신호전달 경로에 대한 ALM16의 영향을 확인한 결과, ALM16은 NF-κB의 핵으로 이동과 DNA-binding activity를 유의적으로 억제하였으며, JNK와 ERK 특이적으로 인산화를 억제함으로써 MAPK 신호전달 경로 활성을 억제하였다. 이러한 결과를 종합하여 볼 때 ALM16이 MAPK와 NF-κB의 신호전달 경로 억제를 통한 iNOS와 COX-2의 발현을 조절하고, 이로 인하여 NO, PGE2 및 pro-inflammatory cytokines의 생성이 감소하여 염증 반응을 조절하는 능력이 있는 것으로 판단된다.

• 한국미생물·생명공학회지
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• 제48권1호
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• pp.79-89
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• 2020

본 연구는 단풍잎돼지풀 70% 에탄올 추출물의 항염증 활성을 검증하기 위해 수행되었다. 단풍잎돼지풀 70% 에탄올 추출물의 전자공여능 측정과 ABTS⁺ 라디칼 소거능을 측정한 결과, 1,000 ㎍/ml 농도에서 각각 84.1%와 92.5%의 효과를 나타냈고, 수렴활성 측정을 한 결과 1,000 ㎍/ml 농도에서 94.7%의 효과를 보였다. 단풍잎돼지풀 70% 에탄올 추출물의 항염증 효과를 측정하기 위해 lipopolysaccharide (LPS)로 유도된 RAW 264.7세포를 사용하여 효과를 측정하였다. 세포에서 단풍잎돼지풀 추출물의 세포독성을 측정하기 위해 MTT assay를 수행하였다. 그 결과, 500 ㎍/ml 농도에서 90% 이상의 생존율을 보였다. Nitric oxide 생산을 억제하는 효과를 측정한 결과, 단풍잎돼지풀 추출물에서 농도가 증가할수록 NO 생성이 감소되는 효과를 확인하였다. 단풍잎돼지풀 추출물의 단백질 발현효과를 western blot을 통해 25, 50, 100 ㎍/ml 농도에서 측정하였고, 양성 대조군으로 β-actin을 사용하였다. 그 결과, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 단백질 발현효과는 100 ㎍/ml 농도에서 8.6%, 25.1%의 감소됨을 확인하였다. ERK1/2, p38, JNK와 Iκ-Bα의 단백질발현 효과는 인산화를 통해 확인하였고, 농도의존적으로 감소하였음을 확인하였다. mRNA 발현 억제 효과를 RT-PCR을 통해 25, 50, 100 ㎍/ml의 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, LPS로 유도된 대식세포에서 iNOS, COX-2, interleukin (IL)-1β, IL-6, TNF-α의 mRNA 발현 억제 효과는 농도가 증가할수록 발현이 감소됨을 확인하였다. 결론적으로 단풍잎돼지풀 추출물은 염증을 억제할 수 있는 가능성이 있는 항염증 소재의 가능성을 증명하였다.

• 한국식품과학회지
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• 제43권1호
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• pp.65-71
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• 2011

등골나물(Eupatorium japonicum)은 식용으로 이용되어온 식물이지만 생리활성에 대한 연구가 없었다. 따라서 본 연구진은 등골나물 에탄올추출물이 쥐 대식세포인 Raw264.7 세포에 LPS로 유도된 염증 반응에 미치는 영향에 대하여 연구를 수행하였다. 등골나물 꽃 부분에 70% 에탄올을 가하여 얻은 등골나물 에탄올추출물(EJE)을 Raw264.7 세포에 LPS와 함께 0, 1, 2.5, 5, 10 mg/L로 처리하여 세포를 배양하였다. LPS에 의해 생성된 NO 및 $PGE_2$ 분비는 EJE를 처리함에 따라 감소하였고 이 결과는 EJE의 독성에 의한 것이 아님이 증명되었다. Raw264.7 세포에 LPS에 의해 생성된 iNOS, COX-2의 단백질과 mRNA의 발현이 EJE의 농도 의존적으로 억제되었으며, 염증 반응시 생성되는 IL-6, IL-${\beta}$, TNF-${\alpha}$의 mRNA 발현도 등골나물 추출물에 의해 현저히 억제되었다. 더욱이 EJE의 분획물 중 EA와 MC 분획물이 독성이 적으면서 효과적으로 NO의 생성을 억제하는 것으로 나타났다. NO 생성 억제효과가 뛰어난 이들 분획물 내의 생리활성 물질에 대한 연구가 추가적으로 수행되어야 한다고 사료된다. 등골나물 꽃을 추출물로 항염증효과를 연구한 논문이 없고 다른 식용 가능한 천연재료들의 항염증효과와 비교하여(data not shown) EJE 분획물의 항염증효과가 낮은 농도에서 탁월한 점을 미루어 볼 때, 등골나물 추출물은 상당히 낮은 농도에서 뛰어난 항염증 물질을 가진 단일물질을 포함할 가능성이 높다고 사료된다. 따라서 위의 결과는 등골나물 추출물이 독성과 부작용이 적은 염증 치료제로 활용될 수 있는 가능성이 높음을 제시한다.

• 한국식품영양과학회지
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• 제33권8호
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• pp.1237-1245
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• 2004

브라질 기원인 신령 버섯 (A. blazei murill)은 강력한 항암 및 면역강화 작용을 가진 것으로 알려져 있다. 본 연구에서는 신령버섯 수용성 추출물(water extracted A. blazei Murill, WEAB)이 A549 인체 폐암세포 증식에 미치는 영향을 조사하였으며, 증식억제와 연관된 기전 해석을 시도하였다. WEAB가 처리된 A549 세포는 처리 농도 의존적으로 생존율이 감소되었으며, WEAB 처리는 암세포의 다양한 형태적 변형을 유발하였다. Flow cytometry 분석 결과로서 WEAB 처리에 의한 A549 폐암세포의 증식억제는 세포주기 G2/M arrest 및 apoptosis 유발과 직접적으로 연관성이 있음을 알 수 있었다. WEAB가 처리된 암세포에서 전사 및 번역 수준에서 cyclin A 발현의 감소 및 Cdk inhibitor p21 발현의 증가 현상이 관찰되었으나, cyclin B1, Cdk2, Cdc2, Wee1, Cdc25c 및 p53 등의 발현에는 큰 변화가 관찰되지 못하였다. 또한 WEAB의 처리는 COX-2 선택적 발현 저하를 유발하였으나, telomere 조절 관련 유전자들의 발현에는 큰 영향을 주지 못하였다. 이상의 결과는 신령버섯 추출물이 강력한 항암 및 암 예방 효능의 잠재력을 가지고 있음을 의미하며, 이에 관한 지속적인 연구가 필요할 것으로 생각된다.