Piperlongumine (PL) is a natural product found in long pepper (Piper longum ). The pharmacological effects of PL are well known, and it has been used for pain, hepatoprotection, and asthma in Oriental medicine. No studies have examined the effects of PL on bone tissue or bone-related diseases, including osteoporosis. The current study investigated for the first time the inhibitory effects of PL on osteoclast differentiation, bone resorption, and osteoclastogenesis-related factors in RAW264.7 macrophages stimulated by the receptor activator for nuclear factor- κB ligand (RANKL). Cytotoxicity was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and osteoclast differentiation and bone resorption were confirmed by tartrate-resistant acid phosphatase (TRAP) staining and pit formation analysis. Osteoclast differentiation factors were confirmed by western blotting. PL exhibited toxicity in RAW264.7 macrophages, inhibiting osteoclast formation and bone resorption, in addition to inhibiting the expression of osteoclastogenesis-related factors, such as tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Fos, and NFATc1, in RANKL-stimulated RAW264.7 macrophages. These findings suggest that PL is suitable for the treatment of osteoporosis, and it serves as a potential therapeutic agent for various bone diseases.

Previous studies demonstrated that rice bran oil has an anti-inflammatory effect. The current study investigated if the immune responses and energy metabolism phenotypes were affected by rice bran oil. Rice bran oil and palm oil, which served as an experimental control, were treated to RAW 264.7 macrophages respectively, followed by LPS stimulation so as to quantify the production of pro-inflammatory cytokines such as IL-6 and TNF-α. The expression of surface markers i.e. CD 80, CD 86 and MHC-classⅡ and mitochondrial metabolism phenotypes were also tested. The data exhibited rice bran oil suppressed inflammatory responses and mitochondrial metabolism were modulated by rice bran oil. These results warrant the necessity for the compositional and/or mechanistic studies to elucidate the details.

Coffee is one of the most familiar beverages to modern human adults, but its bio-physiological effect has not been clearly elucidated. It was known that more than one thousand chemicals were included in the ordinary coffee extract. Among them, the caffein and chlorogenic acid (caffeoylquinic acids) are most abundant and have been investigated by many authors so far. In order to know the real cellular effect of whole coffee extract elements, the dialyzed coffee extract (DCE)1) was made to get coffee elements less than 1000 Da molecular weight, which are freely absorable through gastrointestinal tract. It was directly treated in the culture of RAW 264.7 cells, a murine macrophage lineage. RAW 264.7 cells were treated with DCE equivalent to 2.5 cups of coffee (DCE-2.5), DCE-5, and DCE-10 for 12 hours, and their protein extracts were examined by histological observation and immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the proliferation-related proteins depending on the dose of DCE. DCE-2.5 and DCE-5 enhanced the cellular growth of RAW 264.7 cells by increasing the expression of β-actin, PCNA, Ki-67, MPM2, MAX, cMyc, E2F-1, and Rb-1, and by decreasing the expression of MAD and p21. These proliferation-related proteins were rarely affected by DCE-10. DCE-2.5 and DCE-5 induced the cellular proliferation of RAW 264.7 cells by the signaling of E2F-1 and cMyc, respectively, but these cellular effects almost disappeared in DCE-10. Therefore, it was presumed that the low dose of coffee, DCE-2.5 and DCE-5 might be effective for the proliferation of murine macrophages, RAW264.7 cells, contrast to the high dose of coffee, DCE-10. It was also suggested that the low dose of DCE-2.5 and DCE-5 be helpful to increase the innate immunity in vivo by increasing the cell number of macrophages in contrast to the high dose of DCE-10.

Chronic inflammation, which results from continuous exposure to antigens, is one of major reasons for tissue damage and diseases such as rheumatoid arthritis and type 2 diabetes. In this study, we investigated the anti-inflammatory effects of extracts (hexane, CHCl3, MeOH, MeOH/H2O, and H2O) from GW10-45, which is our new cultivar of an edible mushroom Pleurotus ferulae (ASI 2803 and ASI 2778), in RAW264.7 murine macrophages. None of the extracts showed cytotoxicity in RAW264.7 cells and the hexane, CHCl and H extracts reduced nitric oxide (NO) production, an important inflammatory marker, in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Particularly, the extract (CG45) inhibited NO production more than the other extracts did. To elucidate the effects of CG45 on molecular targets involved in proinflammatory responses, we performed western blot analysis. Expression of inducible nitric oxide (iNOS) significantly decreased in LPS and CG45 co-incubated cells compared to that in LPS only-treated cells. Additionally, another protein thatplays a critical role in inflammation, was down-regulated in cells treated with both LPS and CG45. In the nuclear factor (NF)- B pathway, phosphorylation of I Bα decreased in RAW264.7 cells treated with both LPS and CG45. Furthermore, CG45 inhibited the phosphorylation of NF- B in LPS-stimulated RAW264.7 cells. Conclusively, CG45 could suppress proinflammatory responses in LPS-stimulated RAW264.7 cells by down-regulating not only the phosphorylation of NF- B and I Bα but also the expression of iNOS and COX-2 without any cytotoxicity.

울금의 주요 성분인 커큐민과 대두 추출물인 이소플라본의 피부 미용 측면에서 생리활성을 연구하여 화장품 소재로서 가능성 여부를 규명하고자 하였다. 본 연구는 세포실험을 통해 커큐민과 대두 추출물을 피부 세포에 대한 독성 및 항염증을 확인하고, HPLC을 이용하여 대두 추출물의 성분분석과 피부에 적용하였을 때 수분과 유분, 홍반변화를 측정하였다. 본 연구 결과 커큐민이 RAW 264.7 세포에 대한 독성이 적은 것으로 확인되었으며, 항염증에 대한 효과를 확인하였다. 8주 동안 커큐민과 대두 추 출물이 함유된 화장품과 식이를 병행하여 사용하였을 때 수분함량 변화, 유분함량 변화, 홍반 지수의 감 소가 통계적으로 유의미한 변화를 나타내었으며, 여드름 피부에 유의한 효과를 가질 수 있음을 확인하였 다. 따라서 본 연구는 커큐민과 대두 추출물이 화장품 소재로 사용 시 여드름 피부 개선에 효과적인 화 장품 소재로서 활용 가능성이 있을 것으로 사료된다.

Nanoparticles are widely used in various fields such as electronics, medicines and getting focus on the application in food industry for developing intelligent delivery system with bioactive ingredients or functional nutrients. Basic study on possible toxicological effect of food applicable nanoparticles is required for a practical application in food industry. In this study, size-controlled bovine serum albumin (BSA) nanoparticles were prepared by a desolvation method and their cytotoxicity was investigated. BSA nanoparticles were prepared with mean diameters as 115, 137, 159, and 299 nm, then cytotoxicity was evaluated with RAW 264.7 macrophages as in vitro model. Cell viabilities were significantly affected as increasing nanoparticle concentration. Smaller the sizes of nanoparticles, LD50 values were significantly reduced. LD50 values of BSA nanoparticles were 50, 65, 126, and 170 μg/ml, respectively. Nanoparticle was supposed to induce the apoptosis of RAW 264.7 marcrophages and underlying mechanism will be investigated in future. These findings will be used as valuable basement for nanofood development with BSA nanoparticles.

This study was conducted to investigate the bioconversion of ginsenosides as well as anti-inflammatory activities of fresh ginseng Kkakdugi during fermentation. Fresh ginseng Kkakdugi reached proper ripeness, pH 4.30, and acidity 1.69% at 15oC after 10 days. Lactic acid bacteria grew until reaching 1.10×109 CFU/mL after 20 days of fermentation, and β- glucosidase activity increased from 1.154 to 1.885 units/g. The bioconversion of ginsenosides was confirmed based on increased content of Rg3, an aglycone, from 0.13 to 0.17 mg/g during fermentation through HPLC. Fresh ginseng Kkakdugi did not display cytotoxicity up to the concentrations of 80 μg/mL, regardless of ripening period. Nitrite production and expression of inflammation-related proteins, iNOS and COX-2, decreased in a dose-dependent manner regardless of ripening period. From these results, fresh ginseng Kkakdugi showed the bioconversion of ginsenosides to aglycone during the lactic acid fermentation as well as an anti-inflammatory effect through the reduction of NO production and iNOS and COX-2 expression

생강은 다양한 생리활성이 확인됨에 따라 여러나라에서 전통의약재로서 폭넓게 사용되고 있다. 하지만 진저롤 및 그 유도체에서 유래되는 자극적인 향미로 인하여 식품소재로서의 사용에는 다소 어려움이 있는 실정이다. 본 연구에서는 생강을 Lactobacillus plantarum, Leuconostoc mesenteroides, Streptococcus thermophilus, 및 Lactobacillus acidophilus 등과 같은 유산균에 의한 발효를 통하여 생강의 관능특성을 증진시키려 하였다. 발효된 생강은 in vitro에서 전자공여능 및 cyclooxygenase-2 저해활성을 나타내었으며 또한 산화질소의 생성도 억제하였다. 특히 김치유래 유산균으로 발효한 생강추출물인 GLPe와 GLMe는 Raw 264.7 macrophages에서 prostaglandin-E2 의 생성을 60% 이상 저해하였으며 15-lipoxygenase에 대한 저해 활성도 나타내었다. 따라서 GLPe나 GLMe와 같은 발효생강 추출물은 섭취할 때 자극적인 향미로 인한 거부감이 적으며 염증이나 앨러지 등에 의해 야기되는 바람직하지 않은 증상을 일부 완화시킬 수 있는 건강식품소재로서 사용될 수 있음을 알 수 있었다.

The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and ABTS+ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were 217.04±2.98, 156.72±3.90, and 182.88±3.02 mg TAE/g, respectively, while the electron donating abilities at 1,000 μg/mL of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The ABTS+ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at 50 μg/mL of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

Background : Achyranthes japonica Nakai (AJ) is a perennial herb with a wide distribution in East Asia including Korea, China, and Japan, and it is mainly used as a medicinal plant. In Korea, AJ has been widely used to control pain and improve symptoms in OA patients. AJ contains several important phytochemicals such as saponins, inokosterone, ecdysterone, and oleanolic acid bisdesmoside. Methods and Results : The aim of this work was to investigate the antioxidant and anti-inflammatory activities of fermented and ethanol extracts of Achyranthes japonica Nakai (AJ). The extracts showed strong reductive power and nitrite scavenging, hydroxyl radical scavenging, superoxide radical scavenging, and DNA damage prevention activities. Treatment of RAW 264.7 macrophages with AJ inhibited lipopolysaccharide (LPS)-induced NO secretion and iNOS expression without affecting cell viability. AJ also inhibited cyclooxygenase 2 (COX-2) expression, leading to the suppression of COX-2-derived prostaglandin E2 production. These inhibitory effects of AJ were accompanied by reduced production of tumor necrosis factor-α and interleukins (IL)-1β, -6, and -10. Furthermore, AJ suppressed LPS-induced phosphorylation of extracellular signal regulated kinase, c-Jun N-terminal kinase, and p38. Moreover, AJ inhibited malondialdehyde production and myeloperoxidase activity in LPS-stimulated RAW 264.7 cells. Conclusion : The antioxidant activity of plants is closely related to their medicinal properties and is widely used as a parameter to determine the bioavailability of medicinal plants. The antioxidant and biological activities of AJ extracts might be due to the synergistic actions of multiple bioactive compounds. It can be concluded that AJ extracts are a potential source of biologically important drug candidates.

Arabis glabra is a localized common rhizomatous flowering plant, This plant is often used in Korean traditional systems of medicine as a remedy for blood cleaning, detoxification, abscess, gastrospasm, arthritis, contraction and diarrhea. Generally drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of A. glabra using the ethanolic extract and its sub-fractions. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) on RAW 264.7macrophages. Our results indicated that hexane and chloroform fraction significantly inhibited the LPSinduced NO and PGE2 production in the cells. The hexane fraction inhibitory activity for NO tests with IC50 values showed in 21.0 ㎍/㎖. The chloroform fraction inhibitory activity for PGE2 tests with IC50 values showed in 18.0 ㎍/㎖. These efficacy are expected to be able to present the potential for the development of health functional food for the prevention inflammatory diseases because it has sufficient preventive medical possibilities. Further, it is determined that it is necessary to further study the mechanism of cytokine and protein expression associated with inflammation.