• Plant Resources
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• v.6 no.1
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• pp.1-6
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• 2003

Ginseng(Panax ginseng C. A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark (22 $^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.233-237
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• 2008

The essential oil was obtained from the aerial part of Rhododendon mucronulatum Turcz. var. ciliatum Nakai by steam distillation, samples were collected by headspace (HS) and solid-phase microextraction (SPME) methods, and the compositions of the oil were analyzed by gas chromatography-mass spectrometry (GC-MS). Nineteen constituents were identified from the essential oil: 15 carbohydrates, 3 alcohols, and 1 acetates. Major constituents were 2-${\beta}$-pinene (16.1%), camphene (11.9%), ${\delta}$-3-carene (11.4%), d,l-limonene (9.5%), and ${\gamma}$-terpinene (9.5%). By SPME extraction, seventeen constituents were identified: 13 hydrocarbons, 1 alcohol, 1 nitrogen-containing compound, 1 acetate, and 1 amine. Major constituents of the SPME-extracted sample were cam phene (19.6%), 2-${\beta}$-pinene (18.0%), ${\delta}$-3-carene (17.4%), trimethyl hydrazine (9.7%), ${\gamma}$-terpinene (8.5%), and d,l-limonene (5.5%). By HS extraction, thirteen constituents were identified: 11 hydrocarbons, 1 alcohol, and 1 nitrogen-containing compound. Major constituents of the HS-extracted sample were camphene (25.8%), ${\delta}$-3-carene (24.8%), 2-${\beta}$-pinene (20.2%), d,l-limonene (5.4%), tricyclene (5.1%) and trimethyl hydrazine (4.6%). The fragrance of the essential oil was coniferous, balsamic, and woody, and the $IC_{50}$ value of the essential oil was 0.030 ${\mu}g/mg$ in MTT assay using UaCaT keratinocyte cell line.

• The Korea Journal of Herbology
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• v.31 no.6
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• pp.29-35
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• 2016

Objectives : Blood stasis (BS) is related to be caused by blood circulation and stagnation which are cancer, atherosclerosis and hyperlipidemia in traditional medicine. We extracted 3 kinds of BS formula; Seogakjihwang-tang (SGT), Tonggyuhawlhyul-tang (THT), Hyulbuchukeo-tang (HCT). This study was conducted to investigate whether the 3 kinds of herbal formula extracts have inhibitory efficacy association with anti-adipogenesis. Methods : To investigate the anti-adipogenesis, we used the mouse fibroblast cell line, 3T3-L1 which differentiated into adipocytes in response to insulin, IBMX and dexamethasone (MDI). Cytotoxicity of herbal formula extracts were examined by CCK-8 kit. Intracellular lipid droplets were detected by Oil-Red-O staining. Triglyceride (TG) and leptin were measure using elisa kit. Results : The yield of water extracts was 14.62% (SGT), 21.27% (THT), 20.02% (HCT). Lipid accumulation was reduced significantly by 3 kinds of herbal formula compared to control. Especially, THT and HCT decreased lipid droplet, respectively at all concentration. The TG and leptin were also inhibited by 3 kinds of herbal formula. The IC50 of TG were $280.51{\mu}g/m{\ell}$ (SGT), $52.62{\mu}g/m{\ell}$ (THT), $313.99{\mu}g/m{\ell}$ (HCT). The IC50 of leptin were $348.76{\mu}g/m{\ell}$ (SGT), $164.02{\mu}g/m{\ell}$ (THT), $257.00{\mu}g/m{\ell}$ (HCT). THT was better than other herbal formula on anti-adipogenesis. Conclusion : kinds of herbal formula inhibited adipogenesis in MDI-induced 3T3-L1 adipocytes, as indicated by the significant reduction in TG and leptin concentration without cytotoxicity. Therefore, 3 kinds of herbal formula for BS might act as a therapeutic agent for preventing lipid diseases, such as obesity and atherosclerosis.

• Applied Biological Chemistry
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• v.51 no.1
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• pp.55-59
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• 2008

The composition of essential oil from the aerial part of Erigeron canadensis L. was analyzed by GC-MS. Thirty-one constituents were identified from the essential oil: eighteen hydrocarbons (91.99% of the total oil), two acetates (2.92%), three alcohols (3.59%), four ethers (0.49%), one aldehyde (0.05%), and three ketone (0.23%). Major constituents of the essential oil were D,L-limonene (68.25% of the total oil) and delta-3-carene (15.9%). The $IC_{50}$ value of the essential oil was 0.027 ${\mu}g$ $mg^{-1}$ in MTT assay using HaCaT keratinocyte cell line.

• Molecules and Cells
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• v.37 no.10
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• pp.734-741
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• 2014

Histone modifications on major transcription factor target genes are one of the major regulatory mechanisms controlling adipogenesis. Plant homeodomain finger 2 (PHF2) is a Jumonji domain-containing protein and is known to demethylate the histone H3K9, a repressive gene marker. To better understand the function of PHF2 in adipocyte differentiation, we constructed stable PHF2 knock-down cells by using the mouse pre-adipocyte cell line 3T3-L1. When induced with adipogenic media, PHF2 knock-down cells showed reduced lipid accumulation compared to control cells. Differential expression using a cDNA microarray revealed significant reduction of metabolic pathway genes in the PHF2 knock-down cell line after differentiation. The reduced expression of major transcription factors and adipokines was confirmed with reverse transcription- quantitative polymerase chain reaction and Western blotting. We further performed co-immunoprecipitation analysis of PHF2 with four major adipogenic transcription factors, and we found that CCATT/enhancer binding protein (C/EBP)${\alpha}$ and C/EBP${\delta}$ physically interact with PHF2. In addition, PHF2 binding to target gene promoters was confirmed with a chromatin immunoprecipitation experiment. Finally, histone H3K9 methylation markers on the PHF2-binding sequences were increased in PHF2 knock-down cells after differentiation. Together, these results demonstrate that PHF2 histone demethylase controls adipogenic gene expression during differentiation.

• The Korean Journal of Community Living Science
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• v.28 no.1
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• pp.29-43
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• 2017

We studied the antioxidant, the anti-inflammatory, antithrombotic, and antiobesity activities of seven different kinds of edible plants. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ABTS radical scavenging activities were determined as a measurement of antioxidant activity. NO production inhibition by the macrophage cell line (Raw 264.7) treated with lipopolysaccharide (LPS) was carried out to assess anti-inflammatory activity. Thrombin inhibitory activity was measured for its antithrombotic function and inhibition of 3T3-L1 cell differentiation was evaluated as a measurement of antiobesity activity. Total phenolic components and total flavonoid contents were measured to determine functional materials in medicinal plants. Common sage, Japanese lady bell, and hairy agrimony showed high antioxidant activity ($IC_{50}$) of less than $100{\mu}g/mL$. All samples used in this study showed anti-inflammatory activity. Common sage, hairy agrimony, and hooker chives showed antithrombotic effects. Hairy agrimony showed the highest antithrombotic effect (98.1%). Common sage, Japanese lady belly, hairy agrimony, and hooker chives showed reduced 3T3-L1 cell differentiation and hooker chives strongly inhibited lipid accumulation in the cells compared to other medicinal plants. Common sage and hairy agrimony contained more than 1 mg GAE/g of phenolic compounds and more than 1 mg CE/g of flavonoids. Functional activities were different by plant part and extraction method from each sample. These results suggest that common sage, Japanese lady belly, hairy agrimony, and hooker chives may be used as healthy food sources with antioxidant, anti-inflammatory, antithrombotic, and antiobesity activities, and appropriate extracting methods from each plant need to be developed.

• Natural Product Sciences
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• v.18 no.3
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• pp.153-157
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• 2012

Obesity, which is characterized by excessive fat accumulation in adipose tissues, occurs by fat absorption by lipase and sequential fat accumulation in adipocyte through adipocyte differentiation. Thus, inhibition of pancreatic lipase activity and adipocyte differentiation would be crucial for the prevention and progression of obesity. In the present study, we attempted to evaluate anti-adipogenic activity of several algae extracts employing preadipocytes cell line, 3T3-L1 as an in vitro assay system. The effects on pancreatic lipase activity in vitro were also evaluated. Total methanolic extracts of Cladophora wrightiana and Costaria costata showed significant inhibitory activity on adipocyte differentiation as assessed by measuring fat accumulation using Oil Red O staining. Related to pancreatic lipase, C. wrightiana and Padina arborescens showed significant inhibition. Further fractionation of C. wrightiana, which showed the most potent activity, suggested that $CHCl_3$ and n-BuOH fraction are responsible for adipocyte differentiation inhibition, whereas n-BuOH and $H_2O$ fraction for pancreatic lipase inhibition. Our study also demonstrated that n-BuOH fraction was effective both in early and middle stage of differentiation whereas $CHCl_3$ fraction was effective only in early stage of differentiation. Taken together, algae might be new candidates in the development of obesity treatment.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

• Restorative Dentistry and Endodontics
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• v.21 no.1
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• pp.403-424
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• 1996

Cell culture methods have been used to assess the cytotoxicity of dental materials. Different paramaters are used to monitor cytotoxic effects. But it is difficult to compare each investigator's results with different methods. The objective of this study was to investigate cytotoxic effect of several retrograde filling materials according to cell lines and assay methods. Cytotoxicity of Bestalloy (Dogmyung, Korea), Prisma APH(Densply International Inc., U.S.A.), Clearfil FII (Kuraray Co., Japan), Fuji II (GC Co., Japan), Fuji II LC (GC Co., Japan) and IRM (Densply Co., U.S.A.) on L929, 3T3 and KB permanent cell lines was measured. Radiochromium, Lactate dehydrogenase (LDH) release method and colorimetric assays, namely neutral red (NR) and MTT were used. Each material was mixed according to the manufacturer's instruction. They were tested as solid and extracted state. Cell culture media were added to each mixed or solid materials then the solution was collected and used as extract solutions. Solid Fuji II showed mild cytotoxicity on three cell lines using radiochromium release method. There was no difference in cytotoxicity of extract solution group using radiochromium release method. In colorimetric assay immediate Fuji II group and all the IRM groups showed severe cytotoxic effect. Difference in cyctotoxicity was due to rather kinds of cell lines than assay methods. Solid Fuji II and IRM showed mild cytotoxicity on three cell lines. But extract solutions had different cytotoxic effect according to cell lines using LDH release assay. Light-cured glass ionomer had mild to moderate degree of cytotoxicity on three cell lines. Cytotoxicity was affected by specimen prepaton. Susceptibility of each cell ines were also affected by assay emthods. It was suggested that cytotoxicity study using only one cell line and/or assay method might not accurately reflect the real toxic nature of dental biomaterials.

• Korean Journal of Medicinal Crop Science
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• v.10 no.1
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• pp.51-57
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• 2002

The biological activities of immune modulating activities of the extracts from Echinacea purpurea, Chrysanthmum indicum L. and Circium japonicum var. ussuriense KITAMURA were compared. About 70% of the growth of human hepatocarcinoma and 80% of human gastric cancer cell was inhibited in adding 0.5mg/ml of the ethanol extracts of Echinacea purpurea, Chrysanthmum indicum L. and Circium japonicum var. ussuriense KITAMURA, respectively. The growth of human breast cancer cells was also inhibited in adding 0.5mg/ml of the extracts as well as 60% of the human cancer cells. It was proved that the growth of human normal lung cell, scored as 15% for the extracts. Overall selectivity of the extracts on several human cancer cell line was over 3, which is higher than those from the conventional herbs. The growth of both human immune B and T cells was enhanced up to 1.4 to 2.0 times by adding the extracts, compared to the controls. The secretion of tumor necrosis $factor-alpha(TNF-{\alpha})$ from T cell was also increased up to 94 pg/ml in adding the Echinacea purpurea ethanol extract (0.5mg/ml). Circium japonicum var. ethanol extract also increased up to about 96 pg/ml of interleukin-6(IL-6) from B cell.