To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

The study investigated the potential effect of the Ecklonia cava extract by using Jeju lava seawater (ECE-JLS; hardness: 100, 300 mg/L), which is naturally filtered underground by a volcanic rock layer. The chemical composition, antioxidant-related enzyme activities, radical scavenging activities, reactive oxygen species (ROS) production in Vero cells, and nitric oxide (NO) production in Raw 264.7 cells were measured to evaluate the antioxidant and anti-inflammatory activities of the extract. The total polyphenol and flavonoid contents of ECE-JLS100 and 300 were 156.41± 1.15, 166.16±2.27 mg/g, and 343.76±2.40, 373.90±3.67 mg/g, respectively. The concentration of ECE-JLS’ SOD and CAT-like activities was increased. ECE-JLS in the range of 0.25 to 2 mg/mL exhibited remarkable DPPH radical and hydrogen peroxide scavenging activities. ECE-JLS100 and 300 inhibited total ROS generation by 21.4±0.4 and 23.9± 0.3% in H2O2-induced Vero cells at 200 g/mL concentration, respectively. ECE-JLS100 and 300 decreased NO production, with levels of about 55.0±2.4 and 56.5±1.8% at 200 g/mL concentration, respectively. The contents of TNF-  were decreased compared to the negative control. These observations provide helpful information for the potential industrial use of Jeju lava seawater. Also, ECE-JLSW could be used as a functional food material.

고지방식이를 섭취한 수컷 쥐에서 PPARα activator fenofibrate 단독처방(H/F)과 수영운동 단 독처방(H/S)에 비해 fenofibrate와 수영운동의 조합처방(H/F/S)이 백색지방조직의 염증 개선에 유익한 상 승효과를 나타낼 것인지를 조사하였다. 몸무게, 체내 백색지방조직의 무게 및 혈청 총 콜레스테롤 수치는 저지방식이를 섭취한 쥐(L)에 비해 고지방식이를 섭취한 쥐(H)가 증가하였으며, H에 비해 H/F와 H/S 모 두 감소하였으며, fenofibrate에 의해 감소된 수치는 fenofibrate와 수영운동의 조합처방(H/F/S)에 의해 더 효과적으로 감소하였다. 체내 백색지방조직에서 염증성 사이토카인 유전자와 지방산 산화 관련 유전자의 발현을 조사한 결과, L에 비해 H는 증가하였으며, H에 비해 H/F와 H/S 모두 감소하였고, H/F/S는 H/F 에 비해 더욱 감소시켰다. 따라서 본 연구는 고지방식이를 섭취한 수컷 쥐에서 fenofibrate와 수영운동의 조합처방은 fenofibrate 단독처방에 비해 지방산 산화를 촉진하여 비만으로 발생한 백색지방조직의 염증을 더욱 효과적으로 억제한다는 것을 밝힘으로써, 비만으로 발생하는 지방조직의 염증을 개선하는 실질적인 방법을 제시하였다.

Current studies have revealed the capacity of mesenchymal stem cells (MSCs) in term of immunomodulatory properties, and this distinct potential is downgraded according to the disease duration of patients-derived MSCs. In order to enhance the immunomodulatory and anti-tumorigenic properties of the rheumatoid arthritis (RA) joints-derived MSCs, we aggregate synovial fluid-derived MSCs from RA joints (RA-hMSCs) into 3D-spheroids by the use of hanging drop culture method. Cells were isolated from synovial fluids of RA joints with longstanding active status over 13 years. For aggregation of RA-hMSCs into 3D-spheroids, cells were plated in hanging drops in 30 μL of advanced DMEM (ADMEM) containing 25,000-30,000 cells/ drop and cultured for 48 h. To analyze the comparative immunomodulatory effects of 3D-spheroid and 2D monolayer cultured RA-hMSCs and then cells were cultured in ADMEM supplemented with 20% of synovial fluids of RA patients for 48 h and were evaluated by qRT-PCR for their expression of mRNA levels of inflammatory and antiinflammatory markers. Cellular aggregation of RA-hMSCs was observed and cells were aggregate into a single sphere. Following treatment of RA patient’s synovial fluids into the RA-hMSCs, spheroids formed RA-hMSCs showed significantly (p < 0.05) higher expression of TNFα stimulated gene/protein 6 (TSG-6) than the monolayer cultured RAhMSCs. Therefore, the 3D-spheroid culture methods of RA-hMSCs were more effective than 2D monolayer cultures in suppressing inflammatory response treated with 20% of RA-synovial fluids by expression of TNFα (TSG-6) according to the immune response and enhanced secretion of inflammatory factors.

본 연구에서는 김치로부터 분리한 유산균 Weizmannia coagulans KK7(KCTC19023P) 균주를 이용하여 당근(Daucus carota var. sativa) 잎 추출물의 발효를 실시하고 얻은 생산물의 항염증 활성 및 성 분변화를 확인하였다. 발효에 의한 당근 잎 추출물의 성분 변화를 확인하기 위하여 고성능 액체크로마토그 래피를 이용하여 분석한 결과 발효 전과 비교하였을 때 발효 후에 플라보노이드의 일종인 루테올린 (luteolin)의 함량이 현저히 증가하는 것을 확인할 수 있었다. 당근 잎 추출물과 발효 당근 잎 추출물의 항 염 활성은 LPS에서 염증을 유발한 RAW 264.7 세포에서 NO (Nitric oxide) 생성 억제 효과로 확인하였 다. 발효 당근 잎 추출물을 처리하였을 때 당근 잎 추출물 처리군에 비해 NO 생성이 상대적으로 억제되는 것을 확인할 수 있었다. 발효 당근 잎 추출물 농도가 증가함에 따라 NO 생성 억제 효과도 유의적으로 증 가 하였다. Western blot을 통해 염증반응에서 NO 생합성과 관련된 효소 iNOS의 발현을 확인한 결과 발 효 당근 잎 추출물에서 단백질 발현이 농도 의존적으로 감소하는 것을 확인할 수 있었다. 이러한 실험 결과 로부터 미생물 발효를 통해 당근 잎의 항염 효과가 증대된다는 것을 확인할 수 있었으며 이는 식품으로 거 의 이용되지 않고 버려지는 당근 잎이 발효를 통해 새로운 식품 및 화장품 소재로 사용될 수 있음을 시사 한다.

개똥쑥(Artemisia annua L.)은 국화과 (Compositae)에 속하는 일년생 초본으로 예로부터 진 통, 안면마비 등의 신경계 질환과 위염, 호흡기 등의 질병치료 목적으로 이용되는 식물로 알려져 있다. 개똥쑥을 열수 및 70% 에탄올로 추출하여 추출용매에 따른 추출물의 농도별로 항산화 활성 및 항염증 효과에 미치는 영향을 검토하고자 실시하였다. 총 polyphenol의 함량은 개똥쑥 70% 에탄올 추출물에서 열수 추출물에 비하여 유의적으로 높게 나타났으나, 총 flavonoid 함량은 개똥쑥 열수 추출물과 70% 에 탄올 추출물에서 유의적 차이는 보이지 않았다. DPPH radical 소거능력과 ABTS radical 소거능력은은 62.5∼500 μg/mL 농도에서는 비타민 C 처리구에서 가장 높게 나타났고 그 다음으로는 70% 에탄올 추출물, 열수 추출물 순으로 높게 나타났다. 125 ∼ 500 μg/mL에서 NO 생성 억제효과를 측정한 결 과 열수 추출물에서는 500 μg/mL 농도에서 강한 NO 생성 억제를 보이며, 농도의존적으로 NO 생성 억제를 나타났다. 70% 에탄올 추출물에서는 NO 생성 억제 뿐 아니라 염증성 싸이토카인 발현을 현저 히 억제하는 결과를 보이며, 농도의존적인 억제 효과를 나타났다. 개똥쑥은 항산화효과 및 염증성 싸이 토카인의 발현을 현저히 감소시키는 효과를 보이므로 염증성 질환 치료 및 개선에 널리 사용될 수 있으 리라 사료된다.

The Cheonggukjang used in this study was made with four soybean cultivars grown and harvested at the National Agrobiodiversity Center of the National Academy of Agricultural Sciences: C1 (KLS87248), C2 (Nongrim 51), C3 (GNU- 2007-14613), and C4 (Daewon). The soybeans were inoculated with Bacillus subtilis AFY-2 and fermented for 0, 12, 21, 36, 45, and 60 h in a culture room at 35°C. ABTS and DPPH radical scavenging activities were significantly increased upon fermentation for 12 hours in all samples and maintained or decreased after 21 hour The inhibition of NO production was significantly increased compared to the LPS-treated group, and the C2 sample showed inhibition of NO production at 12 hours of fermentation, and the C3 sample showed inhibition at 21 hours of fermentation. PCA analysis revealed that the ABTS and DPPH radical scavenging activity and NO production inhibitory activity reached peak levels around 12 h fermentation time for all samples. Our results indicate that the optimal fermentation time of the Cheonggukjang sample is 12 hours, confirming the high correlation of all experiments.

The aim of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) isolated from kimchi and antioxidant and anti-inflammatory activities of heat-killed LAB strains. Regarding probiotic properties, Lactiplantibacillus plantarum SMF398 and L. plantarum SMF470 isolated from kimchi can survive under artificial gastric condition and adhere strongly to HT-29 cells. The SMF398 and SMF470 strains showed strong antimicrobial activity and co-aggregation against pathogenic bacteria. The heat-killed cells of LAB (1 mg/mL) were prepared by heating at 121℃ for 15 min. MTT assay revealed that the heat-killed cells (1 mg/mL) of SMF398 and SMF470 were not toxic to HT-29 cells. The heat-killed SMF398 and SMF470 showed significantly higher DPPH and ABTS radical scavenging activities as well as β- carotene bleaching inhibitory activity than the heat-killed L. plantarum ATCC14917, a control probiotic strain (p<0.05). In lipopolysaccharide-induced RAW264.7 cells, the heat-killed SMF398 and SMF470 significantly reduced the nitric oxide production by 30.92% and 22.81%, respectively (p<0.05). Furthermore, the heat-killed SMF398 and SMF470 significantly decreased the gene expression levels of tumor necrosis factor-α, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2 up to 58.02%, 67.70%, 53.44%, 45.90%, respectively (p<0.05). These results suggest that the heat-killed L. plantarum SMF398 and L. plantarum SMF470 might be useful as antioxidant and anti-inflammatory agents.

This study aimed to investigate the effects of antioxidant and anti-inflammatory activities of heat-killed lactic acid bacteria (LAB) produced under different temperature conditions. Regarding probiotic properties, Limosilactobacillus fermentum SMF743 and Lactiplantibacillus plantarum SMF796 isolated from kimchi showed strong acid and bile salt resistance, adhesion activity onto HT-29 cells, and antimicrobial activity against pathogenic bacteria. Based on the results of thermal death time and temperature, heat-killed LAB cells (1 mg/mL) were prepared by heating at 70oC (180 min), 80oC (120 min), 100oC (30 min), and 121oC (15 min). The heat-killed SMF743 and SMF796 showed significantly higher DPPH and ABTS radical scavenging activities than live cells (p<0.05). The heat-killed SMF743 and SMF796 at 70oC or 121oC revealed stronger DPPH and ABTS radical scavenging activities and inhibition of nitric oxide production than those at 80oC or 100oC. Furthermore, heat-killed SMF743 and SMF796 at 121oC significantly reduced the gene expression levels of tumor necrosis factor-, inducible nitric oxide synthase, and cyclooxygenase- 2 up to 53.33%, 58.67%, and 83.67%, respectively, in lipopolysaccharide (LPS)-induced RAW264.7 cells (p<0.05). These results suggest that heat-killed L. fermentum SMF743 and L. plantarum SMF796 can be used as natural antioxidants and anti-inflammatory agents.

Cordyceps militaris mycelium extracts containing high amounts of cordycepin were evaluated in vitro for their antiinflammatory and tumor cell growth-inhibitory activities. All extracts dose dependently inhibited the increased production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), and β-hexosaminidase in lipopolysaccharide (LPS)-stimulated inflammatory cells. All extracts were evaluated for anti-proliferative activity against normal RBL-2H3 cells and diverse types of cancer cell lines, including HCT, MC5-7, U-87MG, AGS, and A549 cells. The extract showed the strongest growth inhibition (IC50 = 28.13 μg/mL) relative to vehicle-treated control cells against fibrosarcoma (MC5-7). We have demonstrated anti-inflammatory activity of C. militaris via inhibition of NO, ROS production, and β-hexosaminidase release in activated cells. C. militaris mycelium extract was also evaluated mechanistically and found to exert six types of anti-cancer activity, confirming its pharmacological potential. Our study suggests C. militaris use as a potential source of anti-inflammatory and anticancer agents. C. militaris may also be considered a functional food.

본 연구에서는 쥐참외뿌리 추출물의 항산화 활성 및 미백, 항염증 활성에 대한 생리활성 효과와 기능성 소재로써의 활용 가능성을 확인해 보고자 하였다. 쥐참외뿌리 추출물의 DPPH 라디칼 소거 활성을 통한 항산화 활성, 멜라닌 세포인 B16F10 melanoma 세포에 대한 멜라닌 생성 억제능을 통한 미백 활성 효과, 대식 세포인 RAW 264.7 세포에 대한 NO 생성 억제능을 통한 항염증 활성 효과를 확인해 보고자 하였다. 연구결과, 쥐참외뿌리 추출물은 농도 의존적으로 DPPH 라디칼 소거활성이 확인되었으며, 양성 대 조군인 Ascorbic acid와 비슷한 DPPH 라디컬 소거활성이 확인되었다. 100 nM α-MSH로 유도된 멜라닌 생성 억제 활성과 LPS 1 ㎍/mL로 유도된 NO 생성 억제능을 유의하게 억제 하는 것을 확인되었다. 이에 따라 쥐참외뿌리 추출물은 항산화 및 미백, 항염증 효과를 가진 기능성 소재로써의 활용 가능성이 있음이 사료되어 진다.

The purpose of this study was to analyze the anti-inflammatory effect of Chung-Dae Indigo Pulverata Levis, indigo naturalis) produced during indigo dyeing. As a result of in vitro cytotoxicity experiments using RAW 264.7 cell, Chung-Dae extract did not inhibit cell proliferation in Raw 264.7 cells in the range of 1~32 μg/mL. NO production was significantly reduced when Chung-Dae extracts were treated at concentrations of 2, 8, and 32 μg/mL (p<0.05). The pro-inflammatory cytokines TNF-α, IL-6, IL-1β and IFN-γ significantly decreased when the Chung-Dae extract was treated at concentrations of 2, 8, and 32 μg/mL compared to the LPS group, and similarly, the TNFα and IL-6 mRNA levels also decreased. Additionally, the mRNA level of COX-2 was also suppressed. At the protein expression level, the expression of TNF-α, IL-6, iNOS and COX-2 were observed with LPS and Chung-Dae extract significantly decreased compared to the group treated with only LPS (p<0.05). From the above results, it shows that Chung-Dae extract, a plant-derived compound, inhibits the inflammatory response induced by LPS in RAW 264.7 cells. and in particular, regulates the inflammatory response by inhibiting the expression of pro-inflammatory cytokines and inflammation-related enzymes.

백출(Atractylodes japonica)은 국화과(Compositae)에 속하는 다년생 식물로 한국에서 이뇨 작용, 수렴작용, 항알러지, 신경보호, 항암, 면역증강 및 위장관 보호효과 등으로 민간요법에서 널리 사 용되고 있다. 백출의 열수 및 70% 에탄올 추출물에 대한 DPPH 및 ABTS 자유라디칼 소거능으로 항산 화 활성을 측정하였다. 게다가 NO 생성, iNOS와 COX-2 단백질 발현을 웨스턴 블롯으로 측정하였으 며, 염증성 사이토카인의 발현을 RT-PCR 방법으로 측정함으로 항염증 효과를 측정하였다. 열수 및 70% 에탄올 추출물들의 항산화 활성은 농도-의존적인 효과를 보였지만, 추출용매에 따른 차이는 나타 나지 않았다. 70% 에탄올에 의한 백출 추출물에서 매우 강한 NO 활성 억제효과를 보였다. 백출 열수 및 에탄올 추출물은 NO 생성에 관여하는 iNOS 및 COX-2 단백질의 발현을 현저히 감소시켰다. 열수 및 70% 에탄올 추출물 모두 염증성 사이토카인인 IL-6 및 IL-1b는 현저히 억제시키지만, TNF-α의 발현은 약하게 억제시키는 결과를 얻었다. 본 연구를 통하여 백출 70% 에탄올 추출물이 NO 생성을 현 저히 억제하며, 염증성 사이토카인의 발현을 현저히 감소시키는 효과를 보이므로 염증성 질환 치료 및 개선에 널리 사용될 수 있으리라 생각한다.