This study was conducted to provide basic data on the antioxidant activity, inhibition of adipocyte differentiation and reactive oxygen species (ROS) production of a mixture of Brassica juncea extract (BJE) and fermented black rice fraction (BRF). We investigated the total phenol content, total flavonoid content, antioxidant effects (DPPH radical scavenging, ABTS radical scavenging, reducing power, FRAP and ORAC assay) and anti-obesity activity of the mixture in 3T3-L1 cells. Our results showed that the total phenol and flavonoid content increased with increasing BRF mixture ratio. The antioxidant activity increased as the BRF mixture ratio increased. In addition, BJE and BRF mixtures did not show any cytotoxicity during the 3T3-L1 differentiation period. During adipocyte differentiation, BJE and BRF mixtures significantly inhibited lipid accumulation and ROS production compared to the control group. These results warrant further experiments to develop an anti-obesity functional food using a mixture of BJE and BRF.

Screening of Bacillus subtilis strains capable of producing extracellular hydrolases, including lipolytic enzyme was carried out. Within the scope of this study, total 61 strains were isolated from various domestic specific environmental samples such as seawater, soil, Jeju ranch, hot spring, salt farm, and fermented food and identified as Bacillus subtilis based on 16S rRNA sequences. Among the isolates, 4 strains had extracellular lipase activity. The growth profile of the strains revealed that 4 strains showed well growth at 50℃, 3 strains at 55℃, and pH 7.0. All strains could tolerate salinity up to 5%(w/v), 3 strains up to 10%(w/v), and only one strain showed growth at 15%(w/v). Additionally, amylolytic and proteolytic activities were detected in these strains. The highest lipolytic, amylolytic, and proteolytic activity was detected in Bacillus subtilis YS-YR 5A. These results demonstrate the potential application of extracellular hydrolase-producing Bacillus subtilis strains, especially strain YS-YR 5A, as fermentation starter to enhance the functionality and multiplication of functional natural products in the food/medicine/cosmetics/bio industries.

본 연구는 십자화과 채소인 갓(Brassica juncea) 추출물 을 이용하여 지표성분인 sinigrin의 함량을 분석하고, 내분비계 교란물질 환경호르몬인 비스페놀 A (BPA)로 분화를 유도한 3T3-L1 전구지방세포에서 갓 추출물과 sinigrin 처 리에 대한 지방세포 분화 및 활성산소종(ROS) 생성 억제, 지방 생성 전사인자(PPARγ, C/EBPα, aP2)의 단백질 발현 감소 효능을 평가하였다. 연구 결과에 따르면 HPLC를 이용하여 측정한 갓 추출물 중 sinigrin의 함량은 21.27±0.2 mg/g 인 것으로 나타났다. BPA로 유도된 3T3-L1 전구지방세포에 서 XTT assay 결과 sinigrin 180 μM 및 갓 추출물 300 μg/ mL 농도에서 세포 독성을 보이지 않았으며, 지방세포 분화과정 중 세포 내 지방 축적량과 ROS 생성량을 비교하였을 때 갓과 sinigrin을 처리한 지방세포의 경우 지방축 적량 및 ROS 생성량 모두 유의적으로 감소시키는 것으로 나타났다. 또한, 갓 추출물 및 sinigrin을 처리하였을 때 지 방세포 분화를 조절하는 전사인자 PPARγ, C/EBPα 및 aP2 의 발현이 억제됨을 확인하였다. 이 결과를 통해 갓은 환 경호르몬 비스페놀 A로 유도된 지방세포 내 지방 축적 억제와 더불어 ROS 저감에 효과적으로 작용함을 확인하였다. 향후 sinigrin을 함유한 갓은 BPA로 인한 지질 대사 장애를 예방하는 천연물 유래 기능성 식품 소재로 활용할 가능성이 높은 것으로 기대되며, 추가로 임상 연구 및 작용기전 입증을 위한 in vivo 모델에서의 후속 연구가 진 행되어야 할 것으로 사료된다.

본 연구의 목적은 oleic acid로 지방생성이 유도된 HepG2 세포에서 자색옥수수 색소 1호 포엽 및 속대 추출물이 간 세포 내 지방생성에 미치는 영향을 구명하는 것이다. 자색옥수수 색소 1호 포엽 및 속대 추출물에 의한 HepG2 세포 내 지방 축적의 변화를 확인하기 위하여 배양된 세포에 oleic acid로 지방 축적을 유도하고 추출물에 의한 중성지방생성 억제 효과를 측정하였으며 추출물을 처리하지 않은 대조군과 추출물을 처리한 실험군의 지방합성 및 축적에 관련된 유전자와 단백질 발현량을 RT-PCR과 Western blot을 통하여 측정하였다. Oil Red O와 Nile Red 염색을 통하여 추출물의 처리로 HepG2 세포 내 중성지방 축적이 억제된 것을 확인하였다. RT-PCR에 의하여 mRNA 발현량을 측정한 결과, oleic acid에 의하여 지방 생성이 유도된 대조군에 비하여 모든 추출물 처리군의 SREBP-1c와 SREBP-1a 유전자 발현량이 유의적으로 감소되었다. Western blot을 실시하여 p-AMPK, p-SREBP1, PPARα, FAS 단백질의 발현량을 측정한 결과, 간에서 지질대사에 관여하는 주요 인자인 SREBP1 단백질의 발현은 추출물의 처리 농도에 따라 유의하게 감소하였으며 지방산의 생합성 경로에 관여하는 주요 효소인 FAS의 단백질 발현향은 모든 처리 농도에서 현저하게 감소된 것이 확인되었다. 본 연구 결과는 자색옥수수 색소 1호 포엽 및 속대 추출물이 간세포 내에서 중성지방의 축적을 억제시키고 지질 합성에 관련된 유전자 및 단백질의 발현을 억제시킴으로써 간 세포 내 지질 축적을 완화할 수 있는 기능성 소재로의 활용가치가 높다고 판단된다.

이 연구는 비만이 심장 조직에서 자가포식 관련 단백질 발현에 미치는 영향을 확인하기 위해 고지방 식이(20주)를 통해 비만을 유도한 후 8주간의 트레드밀 운동을 실시하고, 자가포식의 유도, 형성 그리고 자가포식포와 라이소좀 융합단계를 조절하는 단백질의 발현을 확인하였다. 실험동물(SD rat)은 20주 간의 고지방식이(탄수화물: 20%, 지방: 60%, 단백질: 20%)를 통해 비만을 유도하였으며, 8주간의 트레드 밀 운동(주 5일, 하루 30분, 5분; 8m/min, 5분; 11m/min, 20분; 14m/min)을 실시하였다. 집단 구분은 정상식이 비교군(n=10), 고지방식이 비교군(n=10), 고지방식이 운동군(n=10)으로 구분하였다. 8주간의 트레드밀 운동 실시 전과 후에 경구당부하검사를 실시하여 곡선하 면적(area under the curve; AUC)을 산출 하였으며, 공복시 인슐린 농도와 포도당 농도를 통해 인슐린 저항성 지표인 HOMA-IR과 체중 당 복부지 방량(abdominal visceral fat/Body weight; AVF/BW)를 산출하여 비교하였다. 또한 심장 조직에서 자가포식 관련 단백질을 분석하여 운동 트레이닝의 효과를 검증하였다. 장기간의 고지방식이를 통해 HFD-CON 그룹에서는 비만이 유도되었으며, ND-CON 그룹에 비해 체중, AUC, HOMA-IR, AVF/BW가 증가되는 것으로 나타났다. 하지만 8주간의 트레드밀 운동을 실시한 HFD-TE 그룹에서는 AUC, HOMA-IR, AVF/BW가 개선되는 것으로 나타났다. 체중의 경우, 감소되는 경향은 있었지만, 통계적으로 유의한 차이는 없었다. 자가포식 유도에 관여하는 mTOR와 AMPK는 비만상황에서 모두 감소되었지만, 운동을 통해 증가되는 것으로 나타났다. 자가포식 형성에 관련된 Beclin-1, BNIP3, ATG-7, p62, LC3는 비만상황에서 모두 증가하는 것으로 나타났으며, 운동을 통해 감소되는 것으로 나타났다. 자기포식포와 라이소좀 융합단계 조절하는 Cathepsin L과 LAMP2는 비만상황에서 모두 감소되었으며, 운동을 통해 증가하는 것으로 나타났다. 트레드밀 운동과 같은 신체활동은 대사성 질환에서 나타나는 병리학적 현상을 개선하고 자가포식 과정을 정상적으로 유도하는 것으로 나타났다. 따라서 트레드밀 운동이 심장 관련 질환의 예방 및 치료에 있어 일차적으로 고려해야할 필요성이 있다고 제안한다.

In this paper, we investigate to determine quality characteristics, fatty acid composition and cytotoxic effect of extracts and fractions from whole Lycopus lucidus Turcz. roots. Additionally, we evaluated cytotoxic activity against the growth of human fibrosarcoma cells (HT-1080) and human gastric adenocarcinoma (AGS), human colon cancer cell (HT-29) lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acetone+methylene chloride (A+M) and methanol (MeOH) extracts from L. lucidus Turcz. were obtained through solvent extraction. Then we further fractionated both extracts with n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water. In fatty acid composition, L. lucidus Turcz. contained 33.2% of 18:1n-9 and 1.81% of 18:3n-3, respectively. The incorporation of treatment with A+M and MeOH extracts and n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water fractions dose-dependently increased cytotoxicity against the growth of HT-1080 and AGS, HT-29 cancer cells (p<0.05). The A+M extract had a higher inhibitory effect on the growth of all cancer cells in comparison to MeOH extract. Among the fractions, the 85% aq. MeOH and n-hexane fractions showed a higher inhibitory effect after proliferating the three cancer cells. These results suggest that the 85% aq. MeOH and n-hexane fractions have a potential to inhibit the growth of human cancer cell lines.

In this study, we examined antioxidative effects and the anti-adipogenesis effect of different parts of Cudrania tricuspidata (C), and Morus alba (M). Total polyphenol contents were highest in M-root (34.56±0.045 mg GAE/g), and there was no significant difference, between C-root and M-leaf. Total flavonoid contents of C-root were highest (23.07±0.004 mg QE/g). To examine antioxidant activities of C and M extracts, DPPH and ABTS radical scavenging activity, and FRAP assay, was used. Results show that antioxidant activities of C and M extracts increased, in a dose-dependent manner. Adipocytes are generated by preadipocyte differentiation, during adipogenesis. Matured adipocytes accumulate in abnormal and cause obesity. We investigated effects of leaf and root extracts of C and M, on lipid accumulation, in 3T3-L1 adipocytes. Changes in cell morphology, and degrees of lipid accumulation in adipocytes, were evaluated by Oil Red O staining. Root extracts of C and M, reduced lipid content in a dose-dependent manner. Therefore, root extracts of C and M, may be good candidates for managing obesity.

Purpose: The objective of this study was conducted to investigate the effects of rutin, buckwheat components on cell growth and anti-inflammation in adipocyte 3T3-L1 and human colon cancer cell SW-480. Methods: We cultured 3T3-L1 adipocyte and SW-480 colon cancer cell to confluence, at which time starvation was induced with SFM for 1 day. Cells were then cultured in medium containing 0, 25, 50, or 100 μmol/mL of rutin 3T3-L1 or 0, 10, 20, or 40 μmol/mL SW-480. Cell viability was measured using a cell viability kit. In addition, we examined the expression of mRNA related to inflammation. RT-PCR was used to quantity tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) mRNA levels. Results: Rutin significantly inhibited 3T3-L1 and SW-480 cell proliferation in a dose and time dependent manner. Rutin also significantly reduced the mRNA expression of IL-1β, IL-6 and TNF-α at the highest dose. In addition, rutin treatment caused a significant reduction in COX-2 and iNOS mRNA levels compared to the control group. Conclusion: Overall, our results suggest that rutin has the potential to reduce inflammation, and that these effects are greater during tissue-damaging inflammatory conditions.

Purpose: This study examined the effects of α-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and 400 μmol/L of α-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of α-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The α- lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The α-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, α-lipoic acid inhibited adipocyte 3T3- L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with α-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, (200 μmol/L of α- lipoic acid) treated for 48 hr.

Germinated brown rice (GBR, Orysa sartiva L.) has been reported to have anti-obesity and anti-inflammatory effects. However, the mechanisms underlying these effects in adipocytes are not fully understood. Therefore, this study was conducted to explore the anti-inflammatory mechanisms of GBR on lipopolysaccharide (LPS)-stimulated 3T3-L1 adipocytes. 3T3-L1 adipocytes were pretreated with GBR extracts (0-20 mg/mL) 1 h before LPS stimulation. The mRNA expression of adipokines and Toll-like receptor 4 (TLR4) were measured by RT-PCR. The protein expressions of TLR4- related molecules were detected by western blotting and nuclear factor-κB (NF-κB) activation was measured. Our results showed that GBR extract dose-dependently inhibited mRNA expression of LPS-induced tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1). GBR extract was found to inhibit LPS-induced mRNA expression of TLR4 and protein expression of both myeloid differentiation factor 88 (MyD88) and TNF receptorassociated factor 6 (TRAF6). Furthermore, GBR extract significantly inhibited extracellular receptor-activated kinase (ERK) phosphorylation and NF-κB activation. These results suggest that GBR extract has the anti-inflammatory effects on LPSinduced inflammation via inhibition of TLR4 signaling, includingthe ERK and NF-κB signaling pathways, in adipocytes.

본 연구는 자색옥수수 색소 1호 포엽과 속대 추출물의 항비만 활성을 검정하고자 지방분해효소 저해활성을 평가 하고 3T3-L1 지방전구세포에서 지방분화억제 효과를 검정하고자 수행되었다. Pancreatic lipase 저해 활성 결과, 색소 1호 포엽 및 속대 추출물의 100, 500, 1,000 μg/mL 농 도처리구에서 양성대조군인 orlistat 보다 높은 저해 활성을 나타내었다. 3T3-L1 지방전구세포를 배양하여 색소 1 호 포엽 및 속대 추출물의 세포독성 평가를 수행한 결과, 추출물은 모든 처리농도에서 세포 생존율에 영향을 미치 지 않은 것으로 확인되었다. 분화된 3T3-L1 지방전구세포 에서 색소 1호 포엽과 속대 추출물을 처리하지 않고 분화 시킨 대조군은 lipid droplet의 형성이 활발하게 유발되었으나 색소 1호 포엽 및 속대 추출물의 처리에 의해 농도 의존적으로 lipid droplet의 형성이 억제되는 것으로 나타났다. Real-time PCR과 Western blot을 실시하여 PPARγ와 C/EBPα 유전자 및 단백질 발현량을 측정한 결과, 추출물을 처리하지 않고 분화시킨 대조군에서는 PPARγ와 C/ EBPα의 유전자 및 단백질 발현이 증가하였으며, 추출물 처리에 의해 PPARγ와 C/EBPα의 유전자 및 단백질 발현이 유의적으로 감소하였다. 본 연구 결과는 색소 1호 포엽 및 속대 추출물이 pancreatic lipase 활성 및 지방전구 세포의 분화를 억제시킴으로써 항비만 활성 기능성 물질 로의 활용 가능성이 높음을 시사한다.

The purpose of this study was to evaluate the antioxidant and anti-adipogenic activities of Ligularia stenocephala (L. stenocephala) extract. The contents of the total polyphenol of the extract was 55.950 mg GAE/g residue. Antioxidant activities of L. stenocephala were evaluated by free radical scavenging ability and a reducing power test. 2,2'azino-bis- (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and α-α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging activities of the extract were approximately 90% and 70%, respectively. Reducing power of the extract was 258.833 mg TE/g residue. The anti-adipogenic activity of L. stenocephala extract was examined in 3T3-L1 cells. During adipocyte differentiation, the 3T3-L1 cells were treated both with and without the extract. L. stenocephala extract suppressed the lipid accumulation in a concentration-dependent manner in the 3T3-L1 cells. The L. stenocephala extract inhibited the expression of peroxisome proliferator activated receptor γ (PPARγ) and adipocyte protein 2 (aP2) proteins, compared with control adipocytes. These results indicate that L. stenocephala could be regarded as a potential source natural antioxidant and an anti-obesity agent.

(-)-Epigallocatechin-3-gallate (EGCG) is a major catechin found in green tea. It is reported that EGCG possesses various health benefits including anti-cancer, antioxidant, anti-diabetes, and anti-obesity. The objective of this study was to investigate the effects of EGCG on adipogenesis via activation of AMP-activated protein kinase (AMPK) pathway in 3T3-L1 preadipocytes. In order to determine the effects of EGCG on adipogenesis, preadipocyte differentiation was induced in the presence or absence of EGCG (0~100 μM) for a period of 6 days. EGCG significantly inhibited fat accumulation and suppressed the expression of adipogenic specific proteins including peroxisome proliferator-activated receptor (PPAR)-γ. Also, EGCG markedly increased the activation of AMPK and acetyl-CoA carboxylase (ACC) and the production of intracellular reactive oxygen species (ROS). However, any pretreatment with a specific AMPK inhibitor, compound C, abolished the inhibitory effects of the EGCG on PPARγ expression. This study suggests that EGCG has anti-adipogenic effects through modulation of the AMPK signaling pathway and therefore, may be a promising antiobesity agent.

Aloe-emodin (AE) is the major bioactive component in aloe and known to exhibit anti-inflammatory activities. However, it has not been elucidated whether its anti-inflammatory potency can contribute to the elimination of obesity. The aim of the current study is to investigate the effect of AE on toll-like receptor 4 (TLR4) pathways in the presence of lipopolysaccharide (LPS) in 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with AE (0-20 μM) for one hour, followed by LPS treatment for 30 min and then, adipokine mRNA expression levels were measured. Next, TLR4-related molecules were measured in LPS-stimulated 3T3-L1 adipocytes. AE significantly decreased the mRNA expression of the tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) in a dose-dependent manner. Moreover, AE suppressed TLR4 mRNA expression. Further study showed that AE could suppress the nuclear factor-κB (NF-κB) and phosphorylation of extracellular receptor-activated kinase (pERK). The results of this study suggest that AE directly inhibits TLR4/NF-κB/ERK signaling pathways and decreases the inflammatory response in adipocytes.

The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-α (TNF-α). The lettuce extract suppressed the adhesion of THP-1 to TNF-α-treated HUVEC. The lettuce extract decreased the TNF-α-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism

아이코사노이드는 탄소수 20 개의 다가불포화지방산 산화물로 구성된다. 이들 다가불포화지방산의 생합성 전구물질을 탐지하기 위해 파 밤나방(Spodoptera exigua)의 서로 다른 조직으로부터 지방산을 분리하여 GC/MS로 조성을 분석하였다. 파밤나방 5령 유충에서 소화관, 지방체, 혈구 및 체벽을 분리하고, 각 조직에서 지질을 추출하여 각각 중성지질, 당지질 및 인지질로 분리하였다. 대부분의 조직은 palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2) 그리고 linolenic acid (18:3)를 주요 지방산으로 함유하였다. 그러나 이들 지방산의 조성은 조직과 지질 종류에 따라 상이하였다. 지방체와 혈구세포는 이들 주요 지방산 이외에 myristic acid (14:0)와 3 종류의 미동정 지방산들이 추가로 검출되었다. 서로 다른 지질 종류 가운데 인지질은 중성지질이나 당지질에 비해 상대적으로 높은 linolenic acid를 지닌 반면 포화지방산의 함유량은 낮았다. 전체 불포화지방산의 조성도 조직과 지질 종류에 따라 상이하였다. 인지질은 지방체, 혈구 및 소화관에서 높은 불포화지방산 함유 량을 나타냈다. 세포성 인지질분해효소인 calcium-independent phospholipase A2 (iPLA2)는 지방산 조성을 조절하는 데 역할을 담당하였다. 이 유전자의 RNA 간섭은 중성지질과 인지질에서 지방산 조성의 변화를 유발하였다. 본 연구는 아이코사노이드 생합성의 전구물질로 여겨지는 아라키도닉산을 검출하지 못했다. 이는 곤충에 있어서 아이코사노이드는 포유동물과는 다른 새로운 생합성 과정을 통해 형성되는 것으로 추정된다.