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Inhibition of Adipocyte Differentiation through G1 Arrest by Extract of Sophora tonkinensis Gapnep in 3T3-L1 Preadipocytes (산두근 추출물의 세포주기 정지를 통한 3T3-L1 지방전구세포의 분화 억제)

  • Jeong, Hyun-Young;Hyun, Sook-Kyung;Choi, Yung-Hyun;Kim, Byung-Woo;Kwon, Hyun-Ju
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1346-1353
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    • 2011
  • Sophora tonkinensis Gapnep has been used as a traditional herbal medicine in oriental regions since ancient times. In this study, the effect and mechanism of the MeOH extract of Sophora tonkinensis Gapnep (STME) on adipocite differentiation and adipogenesis in 3T3-L1 preadipocites were investigated. Treatment with STME in the concentration range of 0-200 ${\mu}g$/ml significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner, as determined by a decrease in intracellular lipid droplets and lipid contents measured by Oil Red O staining. In association with the inhibitory effect of lipid accumulation, the expressions of the proteins concerned with adipogenesis in 3T3-L1 preadipocites were also investigated. Treatment with STME reduced the expressions of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ and ${\beta}$ (C/EBP${\alpha}$ and C/EBP${\beta}$) and sterol regulatory element binding protein (SREBP), which are adipocyte specific markers. In flow cytometry analysis, the inhibitory effect of differentiation was caused by G1 arrest and following mitotic clonal expansion cease. Therefore, we also investigated the alteration of G1 phase arrest-related proteins. As a result, the expression of p21 protein was significantly increased, while the expressions of Cdk2, E2F-1 and phospho-Rb were reduced in a dose-dependent manner in STME treated 3T3-L1 cells. According to these results, STME might inhibit differentiation through G1 arrest in 3T3-L1 preadipocytes adipogenesis, and further studies, which are in progress, have to be completed to identify the active compounds.

Association of β-Catenin with Fat Accumulation in 3T3-L1 Adipocytes and Human Population (β-catenin 유전자의 3T3-L1 지방세포 및 인체에서의 지방축적 연관성 연구)

  • Bae, Sung-Min;Lee, Hae-Yong;Chae, Soo-Ahn;Oh, Dong-Jin;Park, Suk-Won;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1301-1309
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    • 2011
  • The major function of adipocytes is to store fat in the form of triglycerides. One of the signaling pathways known to affect adipogenesis, i.e. fat formation, is the WNT/${\beta}$-catenin pathway which inhibits the expression and activity of key regulators of adipogenesis. The purpose of this research is to find genes among the WNT/${\beta}$-catenin pathway which regulate adipogenesis by using small interfering (si) RNA and to find the association of single nucleotide polymorphisms (SNPs) of the gene with serum triglyceride levels in the human population. To elucidate the effects of ${\beta}$-catenin siRNA on adipogenesis key factors, PPAR${\gamma}$ and C/EBP${\alpha}$, we performed real-time PCR and western blotting experiments for the analyses of mRNA and protein levels. It was found that the siRNA-mediated knockdown of ${\beta}$-catenin upregulates adipogenesis key factors. However, upstream regulators of the WNT/${\beta}$-catenin pathway, such as DVL2 and LRP6, had no significant effects compared to ${\beta}$-catenin. These results indicate that ${\beta}$-catenin is a candidate gene for human fat accumulation. In general, serum triglyceride level is a good indicator of fat accumulation in humans. According to statistical analyses of the association between serum triglyceride level and SNPs of ${\beta}$-catenin, -10,288 C>T SNP (rs7630377) in the promoter region was significantly associated with serum triglyceride levels (p<0.05) in 290 Korean subjects. On the other hand, serum cholesterol levels were not significantly associated with SNPs of the ${\beta}$-catenin gene. The results of this study showed that ${\beta}$-catenin is associated with fat accumulation both in vitro and in the human population.

Effects of Organic Acids on In Vitro Ruminal Fermentation Characteristics and Methane Emission (Organic acids 의 첨가가 in vitro 반추위 발효성상과 메탄 생성에 미치는 영향)

  • Ok, Ji Un;Ha, Dong Uk;Lee, Shin Ja;Kim, Eun Tae;Lee, Sang Suk;Oh, Young Kyun;Kim, Kyoung Hoon;Lee, Sung Sill
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1324-1329
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    • 2012
  • The objective of this study was to evaluate the in vitro effects of organic acids on methane emission and ruminal fermentation characteristics. We expected our methodology to result in a decrease of methanogens attached to the surface of rumen ciliate protozoa by addition of organic acids and in particular a decrease in methane emission. A fistulated Holstein cow of 650 kg body weight was used as a donor of rumen fluid. Organic acids (aspartic acid, fumaric acid, lactic acid, malic acid, and succinic acid) known to be propionate enhancers were added to an in vitro fermentation system and incubated with rumen fluid. The microbial population, including bacteria, protozoa, and fungi, were enumerated, and gas production, including methane and fermentation characteristics, were observed in vitro. Organic acids appeared to affect the rumen protozoan community. The rumen protozoal popuation decreased with the addition of aspartic acid, fumaric acid, lactic acid, and malic acid. In particular, the methane emission was reduced by addition of lactic acid. The concentration of propionate with all organic acids that were added appeared to be higher than that of the control at 12 h incubation. Addition of organic acids significantly affected rumen bacteria and microbial growth. The bacteria in added fumaric acid and malic acid was significantly higher (p<0.05) and protozoa was significantly lower (p<0.05) than that of the control. Microbial growth with the addition of organic acids was greater than the control after 48 h incubation.

Inhibition of Adipocyte Differentiation by Methanol Extracts of Oenanthe javanica Seed in 3T3-L1 Preadipocytes (돌미나리씨 추출물에 의한 3T3-L1 지방전구세포의 분화 억제)

  • Ji, Hyang Hwa;Jeong, Hyun Young;Jin, Soojung;Kwon, Hyun Ju;Kim, Byung Woo
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1688-1696
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    • 2012
  • Oenanthe javanica has been used as a food source and also in traditional folk medicine for its detoxifying properties and anti-microbial effects since ancient times. In this study, we evaluated the effect and mechanism of O. javanica seed methanol extract (OJSE) on adipocyte differentiation by 3T3-L1 preadipocytes. Under non-toxic conditions, OJSE treatment resulted in a dose-dependent inhibition of lipid droplet generation and triglyceride accumulation by suppressing adipocyte differentiation, which are associated with the decreased expression of key proadipogenic transcription factors including CCAAR/enhancer binding protein ${\alpha}$, ${\beta}$ ($C/EBP{\alpha}$, $C/EBP{\beta}$) and peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$). OJSE also significantly inhibited proliferation and differentiation of 3T3-L1 preadipocytes through G1-phase arrest, indicating that OJSE blocked mitotic clonal expansion during adipocyte differentiation. Investigation of the alteration of G1 phase arrest-related proteins indicated a dose-dependent increase in the expression of p21 and reduction in expression of cyclin E, Cdk2, E2F-1 and phospho-Rb by OSJE. Taken together, these results suggest that OJSE inhibits adipocyte differentiation by blocking the mitotic clonal expansion, which is accompanied by preadipocyte cell cycle arrest.

Gomisin A Inhibits Tumor Growth and Metastasis through Suppression of Angiogenesis (Gomisin A의 신혈관형성 저해를 통한 종양 성장 및 전이 억제 효과)

  • Kim, Do-Yoon;Yu, Ho-Jin;Yoon, Mi-So;Park, Joo-Hoon;Jang, Sang-Hee;Lee, Hwan-Myung
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1224-1230
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    • 2012
  • Cancer chemotherapy drugs command a large share of the market, and the development of new therapeutics with high efficacy and specificity is an active area of study. Recently, the development of cancer therapeutics from natural products targeting angiogenesis has drawn attention due to conventional chemotherapeutics showing serious side effects and resistance in cancer cells. In this study, we investigated the pharmacological efficacy of Gomisin A, an active ingredient of Schizandra chinensis baillon, on tumor growth and metastasis. Administration of Gomisin A at 10 and 100 ${\mu}g/ml$ reduced tumor growth in vivo by $80.5{\pm}8.1%$ and $96.2{\pm}2%$, respectively, compared with positive tumor controls. Treatment of Gomisin A in normal and various tumor cell lines did not exert significant toxicity. Mice treated with Gomisin A at a concentration of 10 and 100 ${\mu}g$/head showed a significant reduction in tumor-induced angiogenesis of $151{\pm}16.9%$ and $98.5{\pm}29.5%$, respectively. Furthermore, tumor metastasis analysis revealed that the administration of Gomisin A at a concentration of 10 and 100 ${\mu}g$/head inhibited tumor metastasis by $13.5{\pm}8.56%$ and $58.3{\pm}9.12%$, respectively. In addition, Gomisin A significantly decreased cell adhesion of the B16BL6 cells to the extracellular matrix. These results demonstrate that Gomisin A inhibits tumor growth via suppression of angiogenesis and tumor metastasis inhibition, without cellular toxicity. The pharmacological efficacy of Gomisin A suggests that it may be a potential candidate for the development of cancer drugs.

A Study on the Gene Expression of Adipogenic Regulators by an Herbal Composition (생약복합물에 의한 지방세포형성 조절자의 유전자 발현 연구)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.5
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    • pp.729-735
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    • 2010
  • In our previous study, it was reported that an herbal mixture, SH21B, inhibits fat accumulation and adipogenesis both in vitro and in vivo models of obesity. SH21B is a mixture composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd, and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). The aim of this study was to investigate the detailed molecular mechanisms of the effects of SH21B on various regulators of the adipogenesis pathway. During the adipogenesis of 3T3-L1 cells, SH21B significantly decreased the expression levels of central transcription factors of adipogenesis, such as peroxisome proliferator-activated receptor (PPAR)$\gamma$ and CCAAT/enhancer binding protein (C/EBP)$\alpha$. To elucidate the detailed molecular mechanism of the anti-adipogenic effects of SH21B, we examined the expression levels of the various pro-adipogenic or anti-adipogenic regulators of adipogenesis upstream of $PPAR{\gamma}$ and C/$EBP{\alpha}$. The mRNA levels of Krox20 and Kruppel-like factor (KLF) 15, which are pro-adipogenic regulators, were significantly down-regulated by SH21B treatment, whereas the mRNA levels of C/$EBP{\gamma}$ and KLF5 were not changed. KLF2 and C/EBP homologous protein (CHOP), which are anti-adipogenic regulators, were significantly up-regulated by SH21B treatment. These results suggest that the molecular mechanism of the anti-adipogenic effect of SH21B involves both the down-regulations of pro-adipogenic regulators, such as Krox20 and KLF15, and the up-regulations of anti-adipogenic regulators, such as KLF2 and CHOP, which results in the suppression of central transcription factors of adipogenesis including $PPAR{\gamma}$ and C/$EBP{\alpha}$.

Effects of Water Temperature and Estradiol-17β on the Sex Ratio and Growth of the Japanese Eel, Anguilla japonica (극동산 뱀장어, Anguilla japonica의 성비와 성장에 미치는 수온 및 estradiol-17β의 효과)

  • Kim, Dae-Jung;Lee, Nam-Sil;Kim, Shin-Kown;Lee, Bae-Ik;Seong, Ki-Baik;Kim, Kyung-Kil
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1454-1459
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    • 2013
  • This study investigated the effects that water temperature and the administration of estradiol-17${\beta}$ (E2) had on the sex ratio and growth of the Japanese eel, Anguilla japonica. Glass eels (total length${\fallingdotseq}$6.5 cm) were differentiated into an E2 group and an E2-free group and then they were reared for about four months at three water temperature levels of $20^{\circ}C$, $24^{\circ}C$, and $28^{\circ}C$. The results showed that the young eels survived normally at the rearing water temperature of ${\geq}24^{\circ}C$, and grew to a mean size of 20 cm (total length). In the E2-free group, temperature was not found to increase the sex ratio (feminizing rates); however, the sex ratio of the E2-administrated group was found to be a little higher at a high temperature ($28^{\circ}C$). The growth of the E2 group was lower than the growth of the E2-free group at $24^{\circ}C$ and the E2 concentration levels in the plasma at $24^{\circ}C$ were found to be significant after the end of the E2 administration period (178 days). Therefore, we thought that long-term administration of E2 must be considered to be the reason for growth decline in spite of the prominent sex ratio effect. Our results indicate that temperature was not related to an increase in the feminizing rate (sex ratio) in the Japanese eel, Anguilla japonica, and other environmental factors (rearing density, salinity, etc.) that have the possibility of inducing ovarian differentiation must be investigated.

ω3-Polyunsaturated Fatty Acids-induced Inhibition of Tumorigenicity and Invasion by Suppression of COX-2/MMPs/VEGF through NF-kB in Colon Cancer Cells (오메가-3 지방산에 의한 COX-2/MMPs/VEGF 억제에 따른 대장암세포의 종양 형성 및 침윤 억제)

  • Shin, Soyeon;Kim, Yong-Jo;Han, Seung-Hyeon;Silwal, Prashanta;Heo, Jun-Young;Jeon, Young-Joo;Park, Seung-Kiel;Kweon, Gi-Ryang;Park, Jong-Il;Lim, Kyu
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1020-1030
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    • 2017
  • Epidemiology studies have reported a reduced incidence of colon cancer among populations that consume a large quantity of ${\omega}3-polyunsaturated$ fatty acids (${\omega}3-PUFAs$) of marine origin. Herein, we demonstrated a mechanism of anticancer action of ${\omega}3-PUFAs$, showing that they suppressed invasion and tumorigenicity in colon cancer cells. Docosahexaenoic acids (DHA) inhibited the cell growth of HT29 cells. This action likely involved apoptosis, given that the DHA treatment increased the cleaved form of PARP and sub G1 cells. Moreover, the invasiveness of HT29 cells was inhibited following DHA treatment, whereas arachidonic acid (AA) had no effect. The levels of Matrix-metalloproteinase-9 (MMP-9) and MMP-2 mRNA decreased after DHA pretreatment. DHA treatment inhibited MMP-9 and MMP-2 promoter activities and reduced VEGF promoter activity. DHA pretreatment also inhibited the activities of prostaglandin-2 (PGE2)-induced MMPs and the VEGF promoter. Cyclooxygenase-2 (COX-2) overexpression increased the activity of MMPs and that of the Vascular endotherial growth factor (VEGF) promoter in HT29 cells, and DHA inhibited NF-kB and COX-2 promoter reporter activities. As shown by in vivo experiments, when mouse colon cancer cells (MCA38) were implanted into Fat-1 and wild-type mice, both the tumoral size and volume were dramatically inhibited in Fat-1 transgenic mice. Furthermore, TUNEL-positive cells increased in tumors from Fat-1 mice compared with wild mice. In immunohistochemistry, the intensity of CD31 in Fat-1 tumors was weaker. These findings suggest that ${\omega}3-PUFAs$ may inhibit tumorigenicity and angiogenesis as well as cancer cell invasion by suppression of COX-2, MMPs and VEGF via the reduction of NF-kB in colon cancer.

Comparison of the Ingredients at Powdered Green Teas Commercialized in Korea and Japan (한일 말차의 성분 비교)

  • Kim, Kee-Sun;Kouzkue, Nobuyuki;Han, Jae-Sook
    • Journal of the Korean Society of Food Culture
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    • v.19 no.2
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    • pp.177-183
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    • 2004
  • Green tea, a leaf of the plant Camellia sinensis, is one of the most consumed traditional oriental beverages. Green tea has been considered a medicine and a healthful beverage since ancient times, but recently it has received a great deal of attention because of its antioxidants like polyphenols. Moreover, green tea contains amino acids, carbohydrates, proteins, chlorophyll, volatile compounds, minerals, and phytochemical components that are essential or helpful to human health. Depending on the manufacturing process, green teas are classified into several types. Among these, powdered green tea can be effective in the absorption of ingredients compare with other types of green tea since we take the beverage with powder itself. In this paper, the contents of general ingredients (moisture, proteins, fat, carbohydrates, and ash), minerals (Fe, Mg, Ca, Na, K, and P), hunter color values, and alcohol insoluble substance were determined in total of six powdered green teas commercialized in Korea and Japan.

A Study on Optimal Site Selection for Automatic Mountain Meteorology Observation System (AMOS): the Case of Honam and Jeju Areas (최적의 산악기상관측망 적정위치 선정 연구 - 호남·제주 권역을 대상으로)

  • Yoon, Sukhee;Won, Myoungsoo;Jang, Keunchang
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.18 no.4
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    • pp.208-220
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    • 2016
  • Automatic Mountain Meteorology Observation System (AMOS) is an important ingredient for several climatological and forest disaster prediction studies. In this study, we select the optimal sites for AMOS in the mountain areas of Honam and Jeju in order to prevent forest disasters such as forest fires and landslides. So, this study used spatial dataset such as national forest map, forest roads, hiking trails and 30m DEM(Digital Elevation Model) as well as forest risk map(forest fire and landslide), national AWS information to extract optimal site selection of AMOS. Technical methods for optimal site selection of the AMOS was the firstly used multifractal model, IDW interpolation, spatial redundancy for 2.5km AWS buffering analysis, and 200m buffering analysis by using ArcGIS. Secondly, optimal sites selected by spatial analysis were estimated site accessibility, observatory environment of solar power and wireless communication through field survey. The threshold score for the final selection of the sites have to be higher than 70 points in the field assessment. In the result, a total of 159 polygons in national forest map were extracted by the spatial analysis and a total of 64 secondary candidate sites were selected for the ridge and the top of the area using Google Earth. Finally, a total of 26 optimal sites were selected by quantitative assessment based on field survey. Our selection criteria will serve for the establishment of the AMOS network for the best observations of weather conditions in the national forests. The effective observation network may enhance the mountain weather observations, which leads to accurate prediction of forest disasters.