Soojung Jin

Inhibition of Adipocyte Differentiation by Methanol Extracts of Oenanthe javanica Seed in 3T3-L1 Preadipocytes

javanica Seed in 3T3-L1 Preadipocytes Hyang Hwa Ji, Hyun Young Jeong, Soojung Jin, Hyun Ju Kwon and Byung Woo Kim* Department of Life Science and Biotechnology, College of Natural Science, Dong-Eui University, Busan 614-714, Korea Blue-Bio Industry RIC, Dong-Eui University, Busan 614-714, Korea Department of Biomaterial Control (BK21 program), Dong-Eui University Graduate School, Busan 614-714, Korea돌미나리(Oenanthe javanica)는 뛰어난 해독작용과 항균작용으로 예로부터 약재로 사용되어 왔으나 돌미나리씨의 활성은 아직 밝혀지지 않았다. 본 연구에서는 3T3-L1 지방전구세포를 사용하여 돌미나리씨 메탄올 추출물(OJSE)의 지방세포분화에 미치는 영향과 그 기전에 대해 조사하였다. 세포독성이 없는 농도(1~200 μg/ml)의 OJSE를 지방세포 분화유도제와 동시에 지방전구세포에 처리하여 분화시킨 후 Oil Red O 염색을 한 결과 세포 내 lipid droplet 생성 및 triglyceride 축적이 OJSE 농도의존적으로 감소되었으며, 지방세포분화과정에 중요한 역할을 하는 지방세포특이적 마커인 CCAAT/enhancer binding proteins α, β (C/EBPα, C/EBPβ) 및 peroxisome proliferator-activated receptor γ (PPARγ)의 발현이 현저하게 저하되었다. 이는 OJSE에 의해 지방세포 분화 관련 전사인자의 발현이 효과적으로 억제되어 지방세포로의 분화가 저해되고 결과적으로 지방세포내 lipid droplet 생성 및 triglyceride 축적이 억제되었다는 것을 시사한다. 또한 OJSE처리에 의해 mitotic clonal expanstion 단계에서의 세포증식이 저해되었으며, 세포주기의 변화를 분석한 결과, OJSE처리에 의해 지방전구세포의 G1 arrest가 유발됨을 확인하였다. 세포주기 관련 단백질 발현을 분석한 결과, OJSE 농도의존적으로 Cdk inhibitor인 p21의 발현이 현저하게 증가되었으며, 반면 cyclin E, Cdk2, E2F-1 및 phospho-Rb의 발현은 저하됨을 알 수 있었다. 이러한 연구 결과들에 의해 OJSE는 지방전구세포의 G1 arrest를 유도하고 지방세포분화 관련 단백질의 발현을 억제하여 지방세포로의 분화를 억제하는 항비만 효능을 갖는 천연 소재임을 확인하였고, 본 연구는 이를 활용한 향후 지속적인 연구를 위한 기초자료로 그 가치가 매우 높을 것으로 생각된다.

Anti-cancer Activity of Osmanthus matsumuranus Extract by Inducing G2/M Arrest and Apoptosis in Human Hepatocellular Carcinoma Hep G2 Cells

Background: Osmanthus matsumuranus, a species of Oleaceae, is found in East Asia and Southeast Asia. The bioactivities of O. matsumuranus have not yet been fully understood. Here, we studied on the molecular mechanisms underlying anti-cancer effect of ethanol extract of O. matsumuranus (EEOM). Methods: Inhibitory effect of EEOM on cell growth and proliferation was determined by WST assay in various cancer cells. To investigate the mechanisms of EEOM-mediated cytotoxicity, HepG2 cells were treated with various concentration of EEOM and analyzed the cell cycle arrest and apoptosis induction by flow cytometry, Western blot analysis, 4,6-diamidino-2-phenylindole (DAPI) staining and DNA fragmentation. Results: EEOM showed the cytotoxic activities in a dose-dependent manner in various cancer cell lines but not in normal cells, and HepG2 cells were most susceptible to EEOM-induced cytotoxicity. EEOM induced G2/M arrest in HepG2 cells associated with decreased expression of cyclin-dependent kinase 1 (CDK1), cyclin A and cylcin B, and increased expression of phospho-checkpoint kinase 2, p53 and CDK inhibitor p21. Immunofluorescence staining showed that EEOM-treated HepG2 increased doublet nuclei and condensed actin, resulting in cell rounding. Furthermore, EEOM-mediated apoptosis was determined by Annexin V staining, chromatin condensation and DNA fragmentation. EEOM caused upregulation of FAS and Bax, activation of caspase-3, -8, -9, and fragmentation of poly ADP ribose polymerase. Conclusions: These results suggest that EEOM efficiently inhibits proliferation of HepG2 cells by inducing both G2/M arrest and apoptosis via intrinsic and extrinsic pathways, and EEOM may be used as a cancer chemopreventive agent in the food or nutraceutical industry.

Curcumin Blocks Naproxen-Induced Gastric Antral Ulcerations through Inhibition of Lipid Peroxidation and Activation of Enzymatic Scavengers in Rats.

Curcumin is a polyphenol derived from the plant Curcuma longa, which is used for the treatment of diseases associated with oxidative stress and inflammation. The present study was undertaken to determine the protective effect of curcumin against naproxen-induced gastric antral ulcerations in rats. Different doses (10, 50, and 100 mg/kg) of curcumin or vehicle (curcumin, 0 mg/kg) were pretreated for 3 days by oral gavage, and then gastric mucosal lesions were caused by 80 mg/kg naproxen applied for 3 days. Curcumin significantly inhibited the naproxen-induced gastric antral ulcer area and lipid peroxidation in a dose-dependent manner. In addition, curcumin markedly increased activities of radical scavenging enzymes, such as superoxide dismutase (SOD), catalase, and glutathione peroxidase in a dose-dependent manner. Specifically, 100 mg/kg curcumin completely protected the gastric mucosa against the loss in the enzyme, resulting in a drastic increase of activities of radical scavenging enzymes up to more than the level of untreated normal rats. Histological examination obviously showed that curcumin prevents naproxen-induced gastric antral ulceration as a result of direct protection of the gastric mucosa. These results suggest that curcumin blocks naproxen-induced gastric antral ulcerations through prevention of lipid peroxidation and activation of radical scavenging enzymes, and it may offer a potential remedy of gastric antral ulcerations.

Anti-oxidative and Anti-cancer Activities of Treculia africana Extract in Human Colon Adenocarcinoma HT29 Cells

Treculia africana Decne는 빵나무종으로 뽕나무과, Treculia 속에 속하는 식물로서, 이 식물의 다양한 부위에서 추출한 물질은 항염증, 항균등의 효과를 가지고 있어 백일해의 치료등 다양한 질환에서 민간요법으로 사용되어 왔다. 하지만 정확한 생리활성에 관한 연구는 보고된 바가 없다. 따라서 본 연구에서는 T. africana Decne 메탄올 추출물(META)을 사용하여 항산화능 및 인체 대장암 세포주인 HT29에 대한 항암활성에 관하여 분석하였다. DPPH radical scavenging activity를 통해 분석한 결과, META의 IC 50 가 4.53 μg/ml로 강력한 항산화능을 보유하였음을 확인하였다. 또한 META 처리에 의해 HT29의 생존율이 감소함과 더불어 IC 50 가 66.41 μg/ml로 강력한 세포사멸효과를 나타냈다. META 처리에 의해 HT29의 subG1 세포비율 및 Annexin V + 세포의 비율이 증가하고, DAPI로 염색된 apoptotic body가 증가하였다. 또한 apoptosis 관련 단백질인 Fas, Bax, cytochrome c의 발현이 증가하였으며, 이는 caspase 3, 8, 9를 활성화 시켜 최종적으로 PARP가 분해되어 apoptosis가 유도되었음을 확인하였다. 이러한 결과들을 통해 META는 강한 항산화 활성과, 대장암세포에서 apoptosis 유도에 의한 높은 항암 활성을 보유한 물질임을 확인하였다.

Anti-adipogenic Activity of Cortex ulmi pumilae Extract in 3T3-L1 Preadipocytes

and Hyun Ju Kwon 1,5 * Blue-Bio Industry RIC, Dong-Eui University, Busan 614-714, Korea, Department of Biotechnology and Bioengineering, College of Engineering, Dong-Eui University, Busan 614-714, Korea, Department of Food Nutrition, College of Human Ecology, Dong-Eui University, Busan 614-714, Korea, Bioport Korea Co., Marine Bio-industry Development Center, Busan 619-912, Korea, Department of Life Science and Biotechnology, College of Natural Science, Dong-Eui University, Busan 614-714, Korea

Anti-oxidant, Anti-inflammatory and Anti-cancer Effect of Methanol Extract of Pogostemon cablin

In the present study, the substance that show anti-proliferation of cancer cells as well as anti-oxidant and anti-inflammatory effect was searched. As a results, the methanol extract of Pogostemon cablin (P. cablin), is a well-known herb for traditional medicine in Korea and China for treating the digestive disorders, less of appetite, vomiting and diarrhea, inhibited the growth of various cancer cells such as A549, HepG2, MCF7 and HT29 cells. Cytotoxic effect of methanol extraction of P. cablin was ex-cellent in A549 cells. P. cablin extract induced cell cycle arrest at G1 phase of A549 in a dose depend-ent manner. And it induced phosphorylation of p38 and Cdc25A and reduced expression of Cdc25A, Cdks, Cyclins and phospho-Retinoblastoma (Rb) proteins. Therefore, P. cablin extract seems to act through the p38 - Cdc25A - Cdk - Cyclin –Rb pathway in A549 cells. In addition, P. cablin extract showed anti-oxidant effect by DPPH free radical scavenging assay and anti-inflammation effect by in-hibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) in RAW 264.7 cells in a dose-dependent manner. Taken together, these results suggest that P. cablin may be used as not only candidate materi-als for anti-cancer, anti-inflammatory and anti-oxidant, moreover, it would be possible utilized in vari-ous health functional food materials.

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells

Induction of G1 Arrest by Methanol Extract of Lycopus lucidus in Human Lung Adenocarcinoma A549 Cells Lycopus lucidus, a herbaceous perennial, is used as a traditional remedy in East Asia, including China and Korea. It has been reported that L. lucidus has anti-allergic effects, inhibitory effects on cholesterol acyltransferase in high glucose-induced vascular inflammation, and anti-proliferative effects in human breast cancer cells. However, the molecular mechanisms of the anti-cancer effects of L. lucidus have not yet been fully determined. In this study, we evaluated the anti-cancer effect and the mechanism of action of L. lucidus in human lung adenocarcinoma A549 cells using methanol extracts of L. lucidus (MELL). MELL treatment showed cytotoxic activity in a dose-dependent manner and induced G1 arrest in A549 cells. The induction of G1 arrest by MELL was associated with the up-regulation of phospho-CHK2 and the down-regulation of Cdc25A phosphatase. In addition, MELL treatment induced decreased expression of G1/S transition-related proteins, including CDK2, CDK4, CDK6, cyclin D1 and cyclin E. MELL also regulated the mRNA expression of CDK2 and cyclin E. On the other hand, the expression of p53 and the cyclin-dependent kinase inhibitor p21 was not induced by MELL. Collectively, these results suggest that MELL may exert an anti-cancer effect by cell cycle arrest at G1 phase through the ATM/CHK2/Cdc25A/CDK2 pathway in A549 cells.