• Applied Biological Chemistry
• /
• v.38 no.6
• /
• pp.590-595
• /
• 1995

To examine the characteristics of antioxidative compounds from Capsella bursa-pastoris, ethanol extracts were separated into five organic solvent fractions; hexane(Fr.H), diethyl ether (Fr.E), ethyl acetate(Fr.EA), butanol (Fr.B), and water(Fr.D) fractions. Fr.B showed the greatest electron donating ability and inhibitory effect on lipid peroxidation. Whereas Fr.E had the most excellent activity in the superoxide radical scavenging activity by xanthine/xanthine oxidase-cytochrome c reduction system. The inhibitory effect of each fraction on xanthine oxidase was also measured. Fr.E had the strongest inhibitory effect on xanthine oxidase and $IC_{50}$ was $5.65\;{\mu}g$. The results indicate that the superoxide radical scavenging activity of Fr.E is caused by the inhibitory effect on radical generating system of xanthine oxidase. Also the order of inhibitory effect on xanthine oxidase was Fr.B

• Korean Journal of Microbiology
• /
• v.41 no.4
• /
• pp.275-280
• /
• 2005

Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

• Journal of Life Science
• /
• v.26 no.2
• /
• pp.174-180
• /
• 2016

Dicumarol is a coumarin derivative isolated from sweet clover (Melilotus alba), and has anti-coagulant activity with the inhibitory activity of NAD(P)H quinone oxidoreductase1 (NQO1). NQO1 catalyzes the two-electron reduction of quinones to hydroquinones. Dicumarol competes with NAD(P)H for binding to NQO1, resulting in the inhibition of NQO1 enzymatic activity. The expression of matrix metalloproteinases (MMPs) has been implicated in the invasion and metastasis of cancer cells. The expression of MMPs is regulated by cytokines and signal transduction pathways, including those activated by phorbol myristate acetate (PMA). However, the effects of dicumarol on metalloproteinase (MMP)-9 expression and activity are not investigated here. This study investigated whether dicumarol inhibits MMP-9 expression and activity in PMA-treated human renal carcinoma Caki cells. Dicumarol markedly inhibited the PMA-induced MMP-9 mRNA expression and MMP-9 activity. NF-κB and AP1 promoter activity, which is important in MMP-9 expression, also decreased in dicumarol-treated cells. Furthermore, dicumarol markedly suppressed the ability of PMA-mediated migration in Caki cells. When the relevance of NQO1 in the dicumarol-mediated inhibitory effect on PMA-induced MMP9 activity was elucidated, knock-down of NQO1 with siRNA was found to have no effect on PMA-induced MMP9 activity, suggesting that the stimulating effect of dicumarol on PMA-induced MMP9 activity is independent of NQO1 activity. Taken together, the present studies suggested that dicumarol may inhibit PMA-induced migration via down-regulation of MMP-9 expression and activity.

• Korean Journal of Community Nutrition
• /
• v.8 no.1
• /
• pp.26-32
• /
• 2003

This study was designed to assess the nutritional iron status and anemia of middle school girls. Three-hundred-fifty- three female subjects in Ulsan metropolitan city were evaluated using a questionnaire, and hematological indices. The average height and weight of the respondents were 157.19 $\pm$ 5.57 cm and 51.06 $\pm$ 9.42 kg respectively. The average Body Mass Index (BMI, $kg/m^2$) was 20.63 $\pm$ 3.23, which was within the normal range. With regard to clinical symptoms, the greatest number of respondents reported that they experienced 'decreased ability to concentrate'. The total caloric intake of each subject was 1743.28 $\pm$ 343.47 kcal(83.01% of the Korean RDA) and the calcium intake was 634.98 $\pm$ 201.43 mg (79.37% of the Korean RDh) . The mean daily intake of iron was 14.76 : 4.36 mg (92.25% of the Korean RDh) and the heme iron intake was 6.12 $\pm$ 2.30 mg, which was 41.5% of the total iron intake. The average hemoglobin (Hb) concentration of the subjects was 13.24 $\pm$ 1.01 g/dl, and the average hematocrit (Hct) level was 37.79 $\pm$ 4.10%. The transferrin saturation {TS (%)} was 19.41 $\pm$ 9.21%, and the ferritin level was 26.26 $\pm$ 18.60 ng/ml. The iron deficiency anemia among the subjects was estimated at 6.1% by using Hb (< 12 g/ml), 20.5% by using Hct (< 36%), 30.8% by using 75 (< 14%) , and 23.1% by using ferritin (< 12 ng/ml) The total iron binding capacity (TIBC) showed a negative correlation with the Hb, iron, ferritin and 75. With regard to the correlation between blood biochemistry and clinical symptoms related to anemia, the Hb concentration was negatively correlated with 'pale face'(p < 0.05) . In addition, the level of iron was significantly and negatively correlated with 'poor memory' (p<0.01) and the ferritin concentration was negatively correlated with 'no appetite '(p < 0.05) ,'pale face (p < 0.05) . These results suggest that the prevalence of iron deficiency among middle school girls is very high; therefore, guidelines on dietary support and nutritional education to improve their dietary iron status should be provided.

• Journal of Korean Medicine for Obesity Research
• /
• v.20 no.2
• /
• pp.78-87
• /
• 2020

Objectives: To expand and provide information on the efficacy of herbal medicines, anti-obesity effects were evaluated. In many studies, plant-derived components with anti-obesity efficacies have been investigated for their potential inhibitory effects on 3T3-L1 preadipocyte cells. The purpose of this study was to investigate the anti-obesity effects of 65 herbal medicine in 3T3-L1 preadipocyte cells. Methods: Preferentially, 3T3-L1 cells were treated with 65 herbal medicines (500 ㎍/mL) during differentiation for 8 days. Next, 3T3-L1 cells were treated with selected herbal medicines at concentrations ranging from 50 to 200 ㎍/mL during differentiation for 8 days. The accumulation of lipid droplets was determined by Oil Red O staining. The expressions of genes related to adipogenesis were measured by reverse transcription polymerase chain reaction and Western blot analyses. Results: Among the 65 kinds of herbal medicines, 13 herbal medicines that been shown to be effective against the accumulation of lipid droplets were selected. Finally, selected Banhasasim-tang and Samhwangsasim-tang showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affecting cell toxicity. In addition, Banhasasim-tang and Samhwangsasim-tang significantly reduced the expression levels of several adipocyte marker genes including peroxisome proliferator activated receptor-γ and CCAAT/enhancer binding protein-α. Conclusions : These results suggest that the ability of Banhasasim-tang and Samhwangsasimtang has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. Banhasasim-tang and Samhwangsasim-tang may be a promising medicine for the treatment of obesity and related metabolic disorders.

• Journal of Life Science
• /
• v.31 no.8
• /
• pp.729-735
• /
• 2021

Type 2 diabetes occurs when there is an abnormality in the tissue's ability to absorb glucose. Glucose uptake and metabolism by insulin are the basic mechanisms that maintain blood sugar. Glucose uptake goes through various signaling steps initiated by the binding of insulin to receptors on the cell surface. In line with the foregoing, the purpose of this study was to investigate the effect of 2,7-phloroglucinol-6,6-bieckol (PHB), an active compound isolated from Ecklonia cava, on glucose uptake in 3T3-L1 adipocytes. Notably, PHB increased glucose uptake in a dose-dependent manner owing to the enhanced glucose transporter type 4 (GLUT4) expression in the plasma membrane of 3T3-L1 adipocytes. These effects of PHB were attributed to the phosphorylation of insulin receptor substrate-1 and protein kinase B (PKB or AKT), as well as to the phosphoinositide 3-kinase (PI3K) activation in the insulin signaling pathway. PHB also stimulated 5' AMP-activated protein kinase (AMPK) phosphorylation and activation. The phosphorylation and activation of the PI3K/AKT and AMPK pathways by PHB were identified using wortmannin (a PI3K inhibitor) and compound C (an AMPK inhibitor). In this study, we showed that PHB can increase glucose uptake in 3T3-L1 adipocytes by promoting GLUT4 translocation to the plasma membrane via the PI3K and AMPK pathways. The results indicate that PHB may help improve insulin sensitivity.

• Advances in nano research
• /
• v.12 no.5
• /
• pp.501-514
• /
• 2022

This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.49 no.3
• /
• pp.264-273
• /
• 2022

Due to the development of properties of adhesive materials currently used in dentistry, the bonding ability between the brackets and the tooth enamel has been greatly improved. In general, in situations where cooperation can be obtained, adhesion of the orthodontic bracket through the conventional three-step process can show excellent bonding strength. However, if it is difficult to expect patient cooperation, as in the pediatric dentistry area, or if moisture isolation is not properly performed, the binding strength that does not reach the expected effect. As a result, various products that simplify the process for adhesion are being developed. The aim of this study was to evaluate and compare the shear bonding strength between the conventional 3-step adhesion system, self-etching primer system and one-step adhesion system that reduces the priming process. A total of 60 human maxillary, mandibular premolars were prepared. Group I (control group) were followed conventional 3-step bonding process. Group II were conditioned with self-etching primer. Group III were etched with 37% phosphoric acid and brackets were bonded with self-priming adhesive. The resultant shear bond strength of each group was measured and an adhesive remnant index (ARI) was recorded. The mean shear bond strength of group I, II, III were 14.69 MPa, 11.21 MPa and 12.21 MPa respectively. Significant differences could only be found between group I, II and group I, III (p < 0.05). The ARI indicated no significant difference among all groups.

• The Korea Journal of Herbology
• /
• v.33 no.2
• /
• pp.69-77
• /
• 2018

Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, $200{\mu}g/m{\ell}$) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated $receptor-{\gamma}$ ($PPAR-{\gamma}$) and CCAAT/ enhancer-binding $protein-{\alpha}$ ($C/EBP-{\alpha}$). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of $PPAR-{\gamma}$,, $C/EBP-{\alpha}$ in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

• The Korea Journal of Herbology
• /
• v.33 no.2
• /
• pp.19-28
• /
• 2018

Objectives : Epimedii Herba has been frequently used in Korean Traditional Medicine to treat impotence, spermatorrhoea, exophthalmos, and forgetfulness. Present study investigated anti-inflammatory effects of Epimedii Herba water extract (EWE) and attempted to elucidate molecular mechanisms involved. Methods : To explore anti-inflammatory effects of EWE, RAW 264.7 cells, a murine macrophage cell line, were pretreated with $10-100{\mu}g/m{\ell}$ of EWE, and then subsequently exposed to $1{\mu}g/m{\ell}$ of lipopolysaccharide (LPS). Levels of nitric oxide (NO), interleukin-6, $interleukin-1{\beta}$, and tumor necrosis $factor-{\alpha}$ were monitored in the medium. Expression levels of inducible nitric oxide synthase and cyclooxygenase-2 were determined by immunoblot and real-time PCR analyses. Signaling pathways related with nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinases were monitored to elucidate molecular mechanisms involved. Finally, the role of three flavonoid compounds in EWE on LPS-mediated NO production were investigated. Results : In conditioned medium, pretreatment of EWE ($100{\mu}g/m{\ell}$) significantly inhibited LPS-stimulated NO and pro-inflammatory cytokine production. In addition, EWE attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by LPS. EWE prevented the phosphorylation and degradation of inhibitory ${\kappa}B{\alpha}$, nuclear translocation of $NF-{\kappa}B$, and DNA binding of $NF-{\kappa}B$, while EWE did not change the phosphorylation of mitogen-activated protein kinases by LPS. Moreover, icariin, icaritin, and quercetin partly, but significantly, inhibited the LPS-stimulated NO production. Conclusions : These results suggest that EWE has an ability to prevent inflammation in macrophages through inhibition of $NF-{\kappa}B$ signaling pathway.