Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2 , Gsr , HO-1, and catalase , were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2 , HO-1 , and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

We investigated the anti-inflammatory effects of shiitake mushroom and kelp (SMK) mixture extracts in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 cells. Treatment of RAW 264.7 cells with LPS significantly increased NO (nitric oxide) production, pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and IL-1β), and inflammation-related genes (COX-2 and inducible nitric oxide synthase (iNOS)). In cytotoxicity testing using RAW 264.7 cells, SMK mixture extracts in the range of 1-16 μg/mL did not inhibit cell proliferation. However, SMK mixture extracts significantly inhibited NO production in a dose-dependent manner (p<0.05). SMK treatment significantly decreased TNF-α, IL-6, IFN-γ, and IL-1β levels compared to the LPS group, and similarly, pro-inflammatory cytokine mRNA levels also decreased. SMK mixture extracts reduced the mRNA expression of COX-2 and iNOS in RAW 264.7 cells compared to LPS (p<0.05). The above results show that SMK mixture extracts suppressed the inflammatory response induced by LPS. In particular, the extracts were shown to regulate the inflammatory response by suppressing the expression of inflammatory cytokines and inflammation-related enzymes.

We investigated the effect of a synthetic complement peptide C3a on the outcome of Brucella abortus 544 infection in a murine macrophage cell line RAW264.7 cell. First, we determined the highest non-cytotoxic concentration of the peptide in the cell line. We also found that the peptide significantly increased the growth of the bacteria at 8 and 24 h. Although the number of bacterial CFU was also elevated at 48 and 72 h, the increases were not significant as compared to controls. We further investigated the effect of C3a peptide on the growth of Brucella by pre-incubating the peptide at various temperatures and found that the effect was reversed at 24 h post-incubation suggesting that incubation of peptide at high temperatures including 65°C or 95°C could inactivate its action. This also could indicate the beneficial effect of high temperature during infection. Although several studies reported the inhibitory effect of different antimicrobial peptides including C3a, the present study preliminarily revealed that it had no positive contribution on the control of B. abortus 544 infection in vitro and indirectly to its receptor, CD88, which belongs to GPCR. Moreover, the encouraged further exploration of the effect of other similar peptides would be performed for the purpose of finding Brucella-host cell interaction for the control of disease progression.

본 연구는 홍삼추출물의 화장품소재로서의 항염증 효과의 가능성을 확인하는 것을 목적으로 한다. 본 연구에서는 홍삼 추출물을 사용하여 항염증에 대한 생물학적 활성평가를 수행하였다. 대식세포인 RAW 264.7 세포에서 시료의 항염증 효과를 평가하기 위해 MTT assay를 이용한 홍삼 추출물의 독성평가와 nitric oxide 생성 저해 활성 및 염증관련 단백질 및 유전자의 발현량을 확인하였다. LPS로 유도된 RAW 264.7 세포 내에서 시료의 nitric oxide 저해활성은 25 μg/ml에서 71.2 %의 우수한 효능을 나타내었으며, western blot 시험결과 iNOS, COX-2 단백질의 발현은 농도 의존적으로 감소하는 것으로 확인하였다. 이러한 실험 결과를 바탕으로 홍삼추출물이 항염 효과를 가진 화장품 소재로서의 가치를 제안할 수 있다.

This study investigated the anti-inflammatory effects of processed (Beopje) curly dock (Rumex crispus L.) in LPS (lipopolysaccharide)-stimulated murine RAW 264.7 cells. The experimental group was classified into five groups : LPS no treatment, CD (curly dock), CD-B (CD processed through Beopje), LPS, LPS+CD-B (LPS+CD processed through Beopje) and LPS+CD (LPS+CD). Treatment of the Raw 264.7 cell lines using LPS led to a significant increase in NO production, pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β), and inflammation related genes (COX-2 and iNOS). Investigation of the inhibitory effects of CD and processed CD on NO production and expression of iNOS and COX-2 was done in LPS-induced RAW 264.7 cells. There was significant inhibition of NO production by LPS+CD and LPS+CD-B in a dose-dependent manner (p<0.05). Particularly, LPS+CD-B exhibited reduced mRNA expression of iNOS and COX-2 and NO production as compared to LPS+CD in Raw 264.7 cell lines (p<0.05). These results may explain some known biological activities of curly dock including the anti-inflammatory effects. CD-B in particular exhibited the highest anti-inflammatory effects of inhibiting production of NO, through the regulation of inflammatory related genes and pro-inflammatory cytokines. These results of Beopje processing might help decrease the anti-biological effects and increase several active substances of curly dock

In order to perform the biological investigation of coffee extract containing different molecules, it would be necessary to develop in vitro experimental system rather than animal experiment. Although the animal experiment treated via oral intake or intravenous injection may disclose the whole systemic effect, the in vitro cell culture experiment would be more convenient to analyze direct cellular effect of coffee extract than animal experiment. Therefore, this study was aimed to develop a dialysis method for the crude coffee extract to perform the biological investigation using murine macrophage cell line, RAW 264.7. First of all, the RAW 264.7 cells treated with dialyzed coffee extract were observed, and subsequently their protein extracts were analyzed by gel filtration chromatography, thin layer chromatography, and immunoprecipitation high performance liquid chromatography (IP-HPLC). Resultantly, it was found that the low dose (20μg/mL) of dialyzed coffee extract, about 5 cups of ordinary coffee drinking for human adult, enhanced the growth of RAW 264.7 cells by increased expression of β-actin and Ki-67, and also induced the anti-inflammatory effect by decreased expression of NFkB, TNFα, and LC3 contrast to the high dose (40μg/mL) of dialyzed coffee extract. The low dose of dialyzed coffee extract produced almost no harmful effect on RAW cell culture for 12 hours, rather than it produced stimulatory effect on RAW cells by increasing the cell number and enhancing the protein expression of β-actin, Ki-67. Therefore, it was thought that the low dose of dialyzed coffee extract is applicable to cell culture experiment without difficult purification procedures of coffee elements. In addition, as the contrast cellular effect between the low and high dose of coffee extract was found in this study, it was also presumed that the low dose of coffee extract may play an important role in the inflammatory reaction of murine macrophages.

염증은 신체 특정 조직의 감염 및 손상에 관한 생체 반응이며, 매개하는 주요 대상은 면역세포이다. 염증은 급성과 만성 염증으로 나뉘며 신체 조직의 감염 및 손상부위의 규모에 따라 구분 할 수 있다. 염증의 범위가 크게 발현되거나 급성염증 형태로 진행되지 않을 때 만성 염증으로 진행되며 대표적인 만성 염증 질환인 장 질환(Inflammatory bowel disease)의 일종인 크론병 (Crohn’s disease)이나 관절질환인 류머티스성 관절염(Rheumatoid arthritis)으로 나타난다. 낮은 수준이기는 하나 비만 역시 염증성 질환으로 분류할 수 있다. 연리초속 식물이 고래에 신장염을 치료하는 민간처방으로 주로 사용됐기에 이에 착안하여 털연리초(Lathyrus palustris)를 이용하여 세포독성과 항염증 활성 효과를 평가하였다. 대식세포인 RAW 264.7 세포에서 염증 유발 인자인 lipopolysaccharide (LPS)로 자극 후 NO와 PGE2 같은 염증 매개 물질들의 억제 효과를 확인하였다. 털연리초 에탄올 추출물을 처리한 후 염증 매개 물질의 저해율(%)을 측정했을 때 NO 및 PGE2 생성을 농도 의존적으로 현저하게 억제하는 농도는 40 ㎍/mL이었으며 특이적으로 PGE2 발현을 74% 이상 강력히 억제함을 확인하였다. 따라서 본 연구결과는 털연리초의 에탄올 추출물이 유의성 있는 항염증 효과를 나타내었고 이러한 생리활성 효과는 예방의학적 소재로서의 가능성을 충분히 제시할 수 있기에 염증 질환의 예방 및 비만 억제를 위한 기능성 건강식품의 개발로 이어질 것으로 기대된다. 또한 염증 과 관련된 사이토카인 물질인 IL-4, IL-13 및 염증 지표 단백질 인 iNOS, COX-2의 억제 메커니즘과 항염증 활성을 나타내는 핵심 성분의 추가적인 연구가 차후 필요할 것으로 판단된다.

Inflammation is the first response of the immune system to infection or irritation in our body. The use of medicinal plants has been widely applied as an alternative source for drug development. One of marine natural resources, the anti-inflammatory effect of Ishige sinicola ethanol extract (ISEE), was evaluated by using LPS-induced RAW 264.7 cell and mice model. As a result, the production of nitric oxide (NO) and pro-inflammatory cytokines (IL-6, IL-1β, TNF-α) were inhibited with increasing concentration of ISEE without any cytotoxicity. Furthermore, ISEE suppressed the expression of not only inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-κB) p65, and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-Jun N-terminal kinase (JNK) in a dose-dependent manner. In mice ear edema test, the formation of edema was reduced at the highest dosage of ISEE and the reduction of the number of infiltrated mast cells was observed in histological analysis. These results indicate that ISEE has a potent anti-inflammatory activity and can be used as a pharmaceutical material for many kinds of inflammatory disease.

해방풍이 가지는 건강 기능성 소재로서의 가치를 확인하고자, 해방풍 부위별 용매추출물의 항산화 활성 및 면역조절 효과를 조사하였다. 해방풍 부위별 용매추출물의 추출수율은 10.73-30.57이며, 총 폴리페놀 및 총 플라보노이드 함량은 해방풍 잎 70% 에탄올 추출물(EL)에서 각각 10.79 g/100 g 및 2.01 g/100 g으로 가장 높은 값을 나타내었다. DPPH radical 및 ABTS radical 소거활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 12.90-84.70% 및 11.78-57.64%로 가장 우수한 radical 소거활성을 나타내었으며, superoxide radical 소거활성 및 FRAP 활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 각각 49.91-84.05% 및 255.38-975.28 μM로 가장 우수한 활성을 나타내었다. 대식세포주인 RAW 264.7 세포에 해방풍 부위별 용매추출물을 처리하였을 때 nitric oxide 생성 억제능을 확인하기 위해 분석한 결과, 추출용매에 따른 nitric oxide 생성량은 70% 에탄올 및 70% 메탄올 추출물에서 가장 낮은 생성량을 보였으며, 해방풍 부위별에 따른 nitric oxide 생성량은 뿌리, 잎 및 씨앗 순으로 낮았다. 그 중에서 해방풍 뿌리 70% 에탄올 추출물(ER)에서 가장 낮은 nitric oxide 생성량을 보여 우수한 nitric oxide 억제 효과를 확인하였으며, 이를 이용하여 기능성 소재, 미용식품 및 화장품 제조용 원료 등 다양하게 활용 가능할 것으로 사료된다.

Background : Geranium Koreanum(GK), a species of the family Geranium is a perennial herb plants. We performed to determine the anti-inflammatory effects in Lipopoly -saccaride induced RAW 264.7 cell of ethanol extracts from Geranium koreanum. Methods and Results : The dried whole plants 50g of G. koreanum was refluxed three times in 0.5 L 70% ethanol for 2hr. The RAW 264.7 cells for anti-inflammatory assay were obtained from the American Tissue Culture Collection(Manassas, VA), and it cultured in DMEM containing 10% Fetal Bovine Serum(FBS) in 5%, CO2 incubator. An anti-inflammatory of G. koreanum measured by Ntrite oxide(NO) production and the protein expression levels of pro-inflammatory proteins such as COX-2 and iNOS, reductions in activation of NF-kB transcription factor. The results showed that G. koreanum was inhibited NO production and exhibited nontoxic in concentration 50~200㎍/㎖. Also, G. koreanum extracts indicate a significant reduction activation of NF-kB transcription factor and inhibition of pro-inflammatory proteins such as COX-2 and iNOS. Conclusion : The above results suggested that G. koreanum extracts expected the antiinflammatory effects and development possibility as nutraceuticals.

한약재 및 식용으로 사용되는 순비기나무, 번행초, 갯방풍의 재배 한계지역과 생리활성 성분의 효능은 다음과 같다. 1. 순비기나무, 갯방풍의 서해안 북방한계지역은 태안반도이며, 동해안 북방한계지역은 강원도 지역으로 확인되었고, 번행초의 서해안 북방한계지역은 전라남도이고 동해안 북방한계지역은 경상북도 울릉군 지역이었다. 2. 자생지가 해안 200 m 이내 모래점토와 점토흙이 혼합된 지역에서 직사광선은 받는 우점식생이 없는 것을 볼 때 재배 지역은 해안지역과 인접한 지역은 재배 가능지역으로 판단된다. 3. 생리활성효능을 RAW 264.7 세포와 HL-60 세포를 이용하여 확인한 결과 유효한 항염효능은 확인하지 못하였고, 항암활성을 검정한 결과 순비기나무, 번행초는 억제활성이 없었고, 갯방풍은 100μg/ml에서 60%, 200μg/ml에서 72%의 증식억제 효과를 확인하였다.

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects of the ethanol extracts of Rubus coreanus Miq. (ERC) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis factor-α (TNF-α), and Interferon-γ (IFN-γ) production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased IFN-γ and TNF-α production. Consistent with these results, the protein level of inducible Nitric Oxide Synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by ethanol extracts of ERC in a dose-dependent manner. These results suggest that ERC may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.