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Establishment of In Vitro Test System for the Evaluation of the Estrogenic Activities of Natural Products

  • Kim, Ok-Soo;Choi, Jung-Hye;Soung, Young-Hwa;Lee, Seon-Hee;Lee, Jae-Hwa;Ha, Jong-Myung;Ha, Bae-Jin;Heo, Moon-Soo;Lee, Sang-Hyeon
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.906-911
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    • 2004
  • In order to evaluate estrogenic compounds in natural products, an in vitro detection system was established. For this system, the human breast cancer cell line MCF7 was stably trans-fected using an estrogen responsive chloramphenicol acetyltransferase (CAT) reporter plas-mid yielding MCF7/pDsCAT-ERE119-Ad2MLP cells. To test the estrogenic responsiveness of this in vitro assay system, MCF7/pDsCAT-ERE119-Ad2MLP cells were treated with various concentrations of 17f3-estradiol. Treatments of 10$^{-8}$ to 10$^{-12}$ M 17$\beta$-estradiol revealed significant concentration dependent estrogenic activities compared with ethanol. We used in vitro assay system to detect estrogenic effects in Puerariae radix and Ginseng radix Rubra extracts. Treat-ment of 500 and 50 $\mu\textrm{g}$/ml of Puerariae radix extracts increased the transcriptional activity approximately 4- and 1.5-fold, respectively, compared with the ethanol treatment. Treatment of 500, 50, and 5 $\mu\textrm{g}$/ml of Ginseng radix Rubra extracts increased the transcriptional activity approximately 3.2-,2.7, and 1.4-fold, respectively, compared with the ethanol treatment. These observations suggest that Puerariae radix and Ginseng radix Rubra extracts have effective estrogenic actions and that they could be developed as estrogenic supplements.

Phelligridin D maintains the function of periodontal ligament cells through autophagy in glucose-induced oxidative stress

  • Kim, Ji-Eun;Kim, Tae-Gun;Lee, Young-Hee;Yi, Ho-Keun
    • Journal of Periodontal and Implant Science
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    • v.50 no.5
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    • pp.291-302
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    • 2020
  • Purpose: The objective of this study was to investigate whether phelligridin D could reduce glucose-induced oxidative stress, attenuate the resulting inflammatory response, and restore the function of human periodontal ligament cells (HPDLCs). Methods: Primary HPDLCs were isolated from healthy human teeth and cultured. To investigate the effect of phelligridin D on glucose-induced oxidative stress, HPDLCs were treated with phelligridin D, various concentrations of glucose, and glucose oxidase. Glucose-induced oxidative stress, inflammatory molecules, osteoblast differentiation, and mineralization of the HPDLCs were measured by hydrogen peroxide (H2O2) generation, cellular viability, alkaline phosphatase (ALP) activity, alizarin red staining, and western blot analyses. Results: Glucose-induced oxidative stress led to increased production of H2O2, with negative impacts on cellular viability, ALP activity, and calcium deposition in HPDLCs. Furthermore, HPDLCs under glucose-induced oxidative stress showed induction of inflammatory molecules (intercellular adhesion molecule-1, vascular cell adhesion protein-1, tumor necrosis factor-alpha, interleukin-1-beta) and disturbances of osteogenic differentiation (bone morphogenetic protein-2, and -7, runt-related transcription factor-2), cementogenesis (cementum protein-1), and autophagy-related molecules (autophagy related 5, light chain 3 I/II, beclin-1). Phelligridin D restored all these molecules and maintained the function of HPDLCs even under glucose-induced oxidative stress. Conclusions: This study suggests that phelligridin D reduces the inflammation that results from glucose-induced oxidative stress and restores the function of HPDLCs (e.g., osteoblast differentiation) by upregulating autophagy.

Preparation and Characterization of PE Liposomes Containing Antibody (항체를 포함하는 Phosphatidylethanolamine 리포좀의 제조와 그 특성)

  • 박성호;신현재양지원최태부
    • KSBB Journal
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    • v.10 no.2
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    • pp.204-211
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    • 1995
  • A target-sensitive liposome was prepared by using a dioleoyl-phosphatidylethanolamine(DOPE) and a palmitic acid coupled antibody(p-IgG). For the preparation of stable PE-liposomes, the key factors such as antibody modification method with palmitic acrid, molar ratio of p-IgG to lipid and the amount of various additives, were examined. The optimum molar ratio of p-IgG to lipid was found to be $2.5{\times}10^{-4}$ and the final concentration of deoxycholate for the stable liposome formation was about 0.09%. Two kinds of target-sensitive liposomes, containing polyclonal anti-SRBC(Sheep Red Blood Cell)-antibody and monoclonal anti-${\beta}$-HCG(Human Chorionic Gonadotropin)-antibody, were successfully prepared. The destabilization of liposomes was examined by measuring the release of calcein entrapped in the liposome vesicles. Calcein was released only when the liposomes were contacted with the specific target cells. The calcein release with non-specific target cells was negligible. From this result, it is clear that p-IgG is indispensible for the maintenance of stable PE-liposome and the calcein release is mainly due to the specific interactions between the liposomes containing antibody and the target cells containing antigen.

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Effects of Dietary Lycopene Supplementation on Antioxidtion in Broiler and Layer (Lycopene의 사료 내 첨가가 육계와 산란계의 항산화에 미치는 영향)

  • Ju, Won-Don;An, Byeong-Gi;Gang, Chang-Won
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2005.11a
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    • pp.17-30
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    • 2005
  • Lycopene is the red-coloured carotenoid predominantly found in tomato fruit and one of the major carotenoids in the diets of North American and Europeans. Interest in lycopene is growing rapidly following the recent publication of its effects as a natural antioxidant and prevention of cardiovascular disease and cancers. Lycopene, a polyene hydrocarbon carotenoid haying 13 double bond, of which 11 are conjugated double bonds in a linear array exhibits a strong antioxidant property almost twice as strong as that of ${\beta}$-carotene. Lycopene has been shown in recent epidemiological and experimental studies to protect against oxidative damage of DNA which plays an important part in development of various cancer. Lycopene also contribute towards reducing the risk of cardiovascular diseases by preventing oxidation of low-density lipoprotein(LDL) cholesterol. This review summarize our knowledge and the current understanding of lycopene in human health as well as the results of experiments we conducted. We conducted experiments for investigating the effects of antioxidant in broiler and the possibilities of production of high quality eggs containing lycopene by the dietary lycopene supplementation with synthetic lycopene or tomato paste. The results shows that thiobarbituric acid reaction substances(TBARS) values in process of LDL oxidation in blood serum of broiler were significantly decreased by dietary lycopene and tomato paste. The dietary lycopene supplementation resulted in improved egg yolk color and in decreased the malondialdehyde (MDA) of egg yolk after 4 wk of storage at room temperature significantly(P<0.05). The dietary tomato paste was more effective in the MDA reduction compared to the lycopene(P<0.05). The contents of lycopene in egg yolk of the lycopene supplementation groups were significantly higher than those of the control group.

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A Study on the General Characteristics and Instrumental Analysis of Natural Omija Extract

  • Sung, Ki-Chun;Kim, Ki-Jun;Kim, Yong-Ryul;Nam, Sang-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.2
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    • pp.225-232
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    • 2013
  • Omija component was known to possess natural odor, taste, color, and various general characteristics. Omija extraction was extracted using ethanol as a solvent. Omija extract showed a red-purple color of some viscous liquid state. Some conclusions from natural Omija extract were obtained as follow. The result of antimicrobial experiment to add Omija extract, the number of microbial population showed negative reaction from 3 days after it cultivated. This phenomenon could confirm that Omija component affected to antimicrobial effect. The result of dyeing experiment to add Omija extract, fiber dyeing effect showed with some ivory color after dyed to cotton and silk. Also, this phenomenon could confirm that Omija component affected to natural dyeing effect from observated dye state with biological microscope(BM). The result of instrumental analysis, inorganic components of K(109.60ppm), Na(3.500ppm), Ca(1.205ppm), Mg(0.900ppm), Li(0.350ppm), Si(0.380ppm), Cu(0.250ppm), Fe(0.125ppm), Zn(0.090ppm), etcs from Omija were ascertained with ICP/OES, and organic components of benzene(10.808), borny lacetate(11.289), phenol(14.183), ${\beta}$-terpinene(15.840), ${\alpha}$-terpinolene(17.616) etcs from Omija were ascertained with GC/MSD.

Effect of Dried Leaf Powders and Ethanol Extracts of Persimmon, Green Tea and Pine Needle on Lipid Metabolism and Antioxidative Capacity in Rats (감잎, 녹차, 솔잎의 건분 및 에탄올추출물이 흰쥐의 지방대사와 항산화능에 미치는 영향)

  • 김은성
    • Journal of Nutrition and Health
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    • v.32 no.4
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    • pp.337-352
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    • 1999
  • The effects of dried leaf powders and ethanol extract of persimmon, green tea and pine needle on lipid metabolism, lipid peroxidation and antioxidative activity were investigated in rats. Forty-nine male Spargue-Dawley rats weighing 107.8$\pm$1.8g were blocked into seven groups according to body weight. Rats were raised for four weeks with diets containing either 5%(w/w) dried leaf powders of one of three different Korea traditional teas, persimmon(Diospyros kaki Thunb), green tea(Camellia Sinensis O.Ktzc)or pine needle(pinus Koreansis Sieb. Et Zucc), or ethanol from equal amounts of each dried tea powder. Food intake, weight gain, food efficiency ratio, and weights of liver, kidney and epididymal fat were significantly higher in the green-tea-powder group, and significantly lower in the pine-needle-powder and pine-needle-extract groups. Persimmon-leaf powder was found to decrease plasma total lipid, triglyceride and cholesterol concentration by increasing fecal total lipid triglyceride and cholesterol excretions. Liver cholesterol concentration was significantly lower in the green-tea and pine-needle-extract groups. Red-blood-cell superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) activities were significantly increased in rats fed green-tea extract. Liver SOD activity was increased in rats fed pine-needle powder or extract, and liver GSH-px activity was increased in rats fed greentea powder. Plasma and liver thiobarbituric acid reactive substance(TBARS) concentration were both decreased in rats fed dried leaf posers or extracts of persimmon or green tea. It is believed that high vitamin E levels in persimmon leaf, and high flavonoid, beta-carotene and vitamin C levels in green tea effectively inhibited lipid peroxidation. In conclusion, persimmon and green tea leaves were effective in lowering lipid levels and inhibiting lipid peroxidation in animal tissue, while pine needles were effective and lowering body weight gain. From these results, persimmon and green tea leaves can be recommended in the treatment and prevention of chronic discorders such as cardiovascular disease, cancer and aging. As ethanol extracts from these teas were also effecitive in lowering tissue lipid levels and inhibiting lipid peroxidation, we recommend the use of discarded tea grounds for this.

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Effect of Sex Steroid Hormones on Bovine Myogenic Satellite Cell Proliferation, Differentiation and Lipid Accumulation in Myotube

  • Lee, E.J.;Bajracharya, P.;Jang, E.J.;Chang, J.S.;Lee, H.J.;Hong, S.K.;Choi, I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.5
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    • pp.649-658
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    • 2010
  • Myogenic satellite cells (MSCs) are adult stem cells that activate and differentiate into myotubes. These stem cells are multipotent as they transdifferentiate into adipocyte-like cells, nerve cells and osteocytes. The effects of steroid hormones ($E_2$ and testosterone) were studied as a further step toward understanding the mechanism of MSCs proliferation and differentiation. In this study, MSCs were grown continuously for 87 days, implying that there may be a group of MSCs that continue to proliferate rather than undergoing differentiation. Isolated MSCs were cultured in Dulbecco's Modified Eagle's Medium supplemented with adult male, female or castrated bovine serum to observe the effect of steroid hormones on MSC proliferation. Cell proliferation was the highest in cultures supplemented with male serum followed by female and castrated serum. The positive effect of male hormone on MSC proliferation was confirmed by the observation of testosterone-mediated increased proliferation of cells cultured in medium supplemented with castrated serum. Furthermore, steroid hormone treatment of MSCs increased lipid accumulation in myotubes. Oil-Red-O staining showed that 17${\beta}$-estradiol ($E_2$) treatment avidly increased lipid accumulation, followed by $E_2$+testosterone and testosterone alone. To our knowledge, this is the first report of lipid accumulation in myotubes due to steroids in the absence of an adipogenic environment, and the effect of steroid hormones on cell proliferation using different types of adult bovine serum, a natural hormonal system. In conclusion, we found that sex steroids affect MSCs proliferation and differentiation, and lipid accumulation in myotubes.

Effects of a specific blend of essential oils on apparent nutrient digestion, rumen fermentation and rumen microbial populations in sheep fed a 50:50 alfalfa hay:concentrate diet

  • Khateri, N.;Azizi, O.;Jahani-Azizabadi, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.3
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    • pp.370-378
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    • 2017
  • Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.

Immobilization of Lactobionic Acid on Polyurethane Films and Their Interaction with Hepatocytes

  • Meng Wan;Jung Kyung-Hye;Kang Inn-Kyu;Kwon Oh Hyeong;Akaike Toshihiro
    • Macromolecular Research
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    • v.13 no.3
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    • pp.257-264
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    • 2005
  • Polyurethanes containing z-Iysine segments in the main chain (PULL) were synthesized from 4,4'-diphe-nylmethyl diisocyanate, poly(tetramethylene glycol), and z-Iysine oligomer as a chain extender. The PULL film was treated first with a $10\%$ HBr-acetic acid solution and subsequently with a saturated sodium bicarbonate aqueous solution to produce a primary amine group on the surface (PULL-N). Lactobionic acid (LA)-immobilized PULL (PULL-L) was prepared by the coupling reaction of the PULL surface amine groups and the LA carboxylic acid groups. The surface-modified PULLs were then characterized by attenuated total reflection-Fourier transform infra-red spectroscopy, electron spectroscopy for chemical analysis, atomic force microscopy, and contact angle goniometry. In the hepatocytes adhesion experiment, the cells poorly adhered to the PULL surface, although they adhered moderately well to the PULL-N surface. On the other hand, the cells adhered well to the PULL-L surface, suggesting the good affinity of the surface $\beta$-galactose moieties for hepatocytes. When hepatocytes were cultured in the presence of epidermal growth factor for 48 h, the cells rapidly aggregated on the PULL-L surface, whereas they aggregated only slowly on the other surfaces. The PULL prepared in this study has the potential to be used as a coating material for the enhancement of hepatocyte adhesion.

Lactobacillus Aggravate Bile Duct Ligation-Induced Liver Inflammation and Fibrosis in Mice

  • Roh, Yoon Seok;Cho, Ara;Cha, Youn-Soo;Oh, Suk-Heung;Lim, Chae Woong;Kim, Bumseok
    • Toxicological Research
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    • v.34 no.3
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    • pp.241-247
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    • 2018
  • Lactobacillus (LAB) have been reported to exert both harmful and beneficial effects on human and animal health. Recently, it has been reported that dysbiosis and bacterial translocation contribute to liver fibrosis. However, the role of Gram-positive LAB in the situation of chronic liver diseases has not been yet elucidated. Liver injury was induced by bile duct ligation (BDL) in LAB or control-administered mice. Liver fibrosis was enhanced in LAB-administered mice compared with control-treated mice as demonstrated by quantification of Sirius-red positive area, hydroxyproline contents and fibrosis-related genes ($Col1{\alpha}1$, Acta2, Timp1, Tgfb1). Moreover, LAB-administered mice were more susceptible to BDL-induced liver injury as shown by increased ALT and AST level of LAB group compared with control group at 5 days post BDL. Consistent with serum level, inflammatory cytokines ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$) were also significantly increased in LAB-treated mice. Of note, LAB-treated liver showed increased lipoteichoic acid (LTA) expression compared with control-treated liver, indicating that LAB-derived LTA may translocate from intestine to liver via portal vein. Indeed, responsible receptor or inflammatory factor (PAFR and iNOS) for LTA were upregulated in LAB-administered group. The present findings demonstrate that administration of LAB increases LTA translocation to liver and induces profibrogenic inflammatory milieu, leading to aggravation of liver fibrosis. The current study provides new cautious information of LAB for liver fibrosis patients to prevent the detrimental effect of LAB supplements.