• Asian Pacific Journal of Cancer Prevention
• /
• 제17권9호
• /
• pp.4267-4273
• /
• 2016

Previous studies suggested that dithio-carbamates are potent apoptosis and anti-apoptosis inducing agents in various cancer cells. Here, the anti-proliferative and apoptosis inducing effects of a new derivative (2-NDC) from the dithio-carbamate family was examined in human leukemia K562 cells. We use thiazolyl blue tetrazolium bromide (MTT) to measure viability and cell growth inhibition. The 2-NDC showed effects on viability in a dose and time-dependent manner, inhibiting proliferation at concentrations of $10-30{\mu}M$ after 24-48 hours of treatment and increasing values after 72 hours at $40-120{\mu}M$. The cytotoxic effect of the compound was calculated with an $IC_{50}$ of $30{\mu}M$ after 24-hour. Apoptosis induction was confirmed by acridine orange-ethidium bromide (AO/EtBr) staining, DNA fragmentation assay, flow cytometric assessment and also caspase-3 activation assay. Furthermore, enzymes level such as superoxide dismutase (SOD) and catalase (CAT) involved in oxidative stress were evaluated. The results of this study demonstrated insignificant increase of intracellular ROS levels for 24 hours and reduction after 48-72 hours. In addition to reduction of intracellular thiol, caspase-3 like activity was also decreased in a time-dependent manner in cells treated with 2-NDC. Thus 2-NDC can be considered as a good candidate for further pharmaceutical evaluations.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권13호
• /
• pp.5371-5376
• /
• 2015

Berberine (B1), isolated from stems of Coscinium fenestratum (Goetgh.) Colebr, was used as a principle structure to synthesize three phenolic derivatives: berberrubine (B2) with a single phenolic group, berberrubine chloride (B3) as a chloride counter ion derivative, and 2,3,9,10-tetra-hydroxyberberine chloride (B4) with four phenolic groups, to investigate their direct and indirect antioxidant activities. For DPPH assay, compounds B4, B3, and B2 showed good direct antioxidant activity ($IC_{50}$ values=$10.7{\pm}1.76$, $55.2{\pm}2.24$, and $87.4{\pm}6.65{\mu}M$, respectively) whereas the $IC_{50}$ value of berberine was higher than $500{\mu}M$. Moreover, compound B4 exhibited a better DPPH scavenging activity than BHT as a standard antioxidant ($IC_{50}=72.7{\pm}7.22{\mu}M$) due to the ortho position of hydroxyl groups and its capacity to undergo intramolecular hydrogen bonding. For cytotoxicity assay against human fibrosarcoma cells (HT1080) using MTT reagent, the sequence of $IC_{50}$ value at 7-day treatment stated that B1 < B4 < B2 ($0.44{\pm}0.03$, $2.88{\pm}0.23$, and $6.05{\pm}0.64{\mu}M$, respectively). Berberine derivatives, B2 and B4, showed approximately the same level of CAT expression and significant up-regulation of SOD expression in a dose-dependent manner compared to berberine treatment for 7-day exposure using reverse transcription-polymerase chain reaction (RT-PCR) assays. Our findings show a better direct-antioxidant activity of the derivatives containing phenolic groups than berberine in a cell-free system. For cell-based system, berberine was able to exert better cytotoxic activity than its derivatives. Berberine derivatives containing a single and four phenolic groups showed improved up-regulation of SOD gene expression. Cytotoxic action might not be the main effect of berberine derivatives. Other pharmacological targets of these derivatives should be further investigated to confirm the medical benefit of phenolic groups introduced into the berberine molecule.

• 대한한방내과학회지
• /
• 제20권1호
• /
• pp.99-110
• /
• 1999

Effects of Chiyangtang(CYT) on H. pylori-induced increase of interleukin 8 and interleukin 1 gene expression was studied in Kato Ⅲ cell line, a human stomach epithelial cell line. Treatment of H. pylori to the cell culture signifant!y increased IL-8 and IL-1 mRNA synthesis. When CYT was added along with H. pylori, the increase of IL-8 and IL-1 mRNA synthesis was blocked. Activation of transcription factor $NF-{\kappa}B$ and AP-1 which were known to important in IL-8 and IL-1 gene expression was also studied using chloramphenicol acetyltransferase(CAT) assay. Treatment of H. pylori increased activation of $NF-{\kappa}B$ and AP-l and CYT effectively protected the activation. Electrophoretic mobility shift assay suggested that CYT effectively inhibited DNA binding of $NF-{\kappa}B$ and AP-l to their cognate site. These results suggested that CYT could prevent stomach diseases through the down regulation of IL -8 and IL-l gene expression which might be mediated by the inhibition of $NF-{\kappa}B$ and AP-1 activities and their binding to DNA.

• Molecules and Cells
• /
• 제38권2호
• /
• pp.187-194
• /
• 2015

Thioredoxin (TRX) is a disulfide reductase present ubiquitously in all taxa and plays an important role as a regulator of cellular redox state. Recently, a redox-independent, chaperone function has also been reported for some thioredoxins. We previously identified nodulin-35, the subunit of soybean uricase, as an interacting target of a cytosolic soybean thioredoxin, GmTRX. Here we report the further characterization of the interaction, which turns out to be independent of the disulfide reductase function and results in the co-localization of GmTRX and nodulin-35 in peroxisomes, suggesting a possible function of GmTRX in peroxisomes. In addition, the chaperone function of GmTRX was demonstrated in in vitro molecular chaperone activity assays including the thermal denaturation assay and malate dehydrogenase aggregation assay. Our results demonstrate that the target of GmTRX is not only confined to the nodulin-35, but many other peroxisomal proteins, including catalase (AtCAT), transthyretin-like protein 1 (AtTTL1), and acyl-coenzyme A oxidase 4 (AtACX4), also interact with the GmTRX. Together with an increased uricase activity of nodulin-35 and reduced ROS accumulation observed in the presence of GmTRX in our results, especially under heat shock and oxidative stress conditions, it appears that GmTRX represents a novel thioredoxin that is co-localized to the peroxisomes, possibly providing functional integrity to peroxisomal proteins.

• Journal of Microbiology and Biotechnology
• /
• 제18권3호
• /
• pp.397-403
• /
• 2008

Nikkomycins are a group of peptidyl nucleoside antibiotics with potent fungicidal, insecticidal, and acaricidal activities. sanN was cloned from the partial genomic library of Streptomyces ansochromogenes 7100. Gene disruption and complementation analysis demonstrated that sanN is essential for nikkomycin biosynthesis in S. ansochromogenes. Primer extension assay indicated that sanN is transcribed from two promoters (sanN-P1 and sanN-P2), and sanN-P2 plays a more important role in nikkomycin biosynthesis. Purified recombinant SanN acts as a dehydrogenase to convert benzoate-CoA to benzaldehyde in a random-order mechanism in vitro, with respective $K_{cat}/K_m$$ values of $3.8mM^{-1}s^{-1}\;and\;12.0mM^{-1}s^{-1}$ toward benzoate-CoA and NADH, suggesting that SanN catalyzes the formation of picolinaldehyde during biosynthesis of nikkomycin X and Z components in the wild-type stain. These data would facilitate us to understand the biosynthetic pathway of nikkomycins and to consider the combinatorial synthesis of novel antibiotic derivatives.

• 생약학회지
• /
• 제35권4호통권139호
• /
• pp.357-363
• /
• 2004

Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiiflammatory, antihistaminic, antioxidant and free-radical scavenging abilities. To determined flavonoid, myricetin in the presence of other antioxidants - vitamin C and vitamin E - can exert antioxidative properties not only directly by modulating the AOE system but also scavenging free radical, we investigated cell viability, antioxidant enzyme activities and ROS level in B16F10 murine melanoma cell. B16F10 cells were exposed to medium containing myricetin in the presence or absence of vitamin C or vitamin E for a period of 24 hr. Cell viability was measured by MTT assay. In co-treating myricetin with other antioxidants, CAT activities were increased, compared with control, but SOD and GPx activities were decreased, compared with each antioxidant treated groups . In the group of myricetin or myricetin present with other antioxidants, ROS levels were decreased dose-dependently. Especially, myricetin present of other antioxidants were decreased compared with myricetin.

• Bulletin of the Korean Chemical Society
• /
• 제26권9호
• /
• pp.1385-1389
• /
• 2005

Human hepatitis B virus (HBV) is a pathogen related to the development of liver diseases including chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). However, the efficient methods to suppress HBV replication have not been developed yet. Therefore, we have used RNA interference (RNAi) as a potential tool for the suppression of HBV replication. Here, we designed a 21 nt small intefering dsRNA (siRNA) against hepatitis B virus X (HBx) RNA with 3' overhanging ends derived from T7 promoter. It has been reported that HBV X protein plays an important role in HBV gene expression and viral replication. The suppression of HBx gene expression by the 21 nt siRNA was investigated by Northern blot analysis and chloramphenicol acetyl transferase (CAT) assay. The level of HBx mRNA was decreased by siRNA in a dose-dependent manner. We also found that the 21 nt siRNA inhibited the HBV replication in hepatocellular carcinoma cell.

• 한국초지조사료학회지
• /
• 제39권4호
• /
• pp.298-302
• /
• 2019

Acidic soil significantly reduces crop productivity mainly due to aluminum (Al) toxicity. Alfalfa (Medicago sativa L.) roots were exposed to aluminum stress (Al₃⁺) in calcium chloride (CaCl₂) solution (pH4.5) and root growth, physiological and antioxidant enzyme responses were investigated. The root growth (length) was significantly inhibited after 48 h of aluminum stress imposition. Histochemical staining with hematoxylin indicated significant accumulation of aluminum in Al stress-treated root tissues. Histochemical assay were also performed to detect superoxide anion, hydrogen peroxide and lipid peroxidation, which were found to be more in root tissues treated with higher aluminum concentrations. The enzymatic activity of CAT, POD and GR in root tissues was slightly increased after Al stress treatment. The result suggests that Al stress alters root growth in alfalfa and induces reactive oxygen species (ROS) production, and demonstrates that antioxidant enzymes involved in detoxification of Al-mediated oxidative stress.

• 약학회지
• /
• 제47권6호
• /
• pp.432-437
• /
• 2003

It has been known that quercetin, a bioflavonoid widely distributed in fruits and vegetables as dietary-derived flavonoid and exert significant multiple biological effects such as antioxidant and anti-inflammatory, anti-tumor effects. In addition, it has been shown to have a chemoprotective role in cancer, though complex effects on signal transduction involved in cell proliferation and angiogenesis. The present study investigated whether quercetin can enhance antioxidant enzyme activity (glutathione peroxidase: GPx, superoxide dismutase: SOD, catalase: CAT) and regulate the reactive oxygen species (ROS) generation in the presence of vitamin E, L-ascorbic acid, reduced glutathione (GSH) on B16F10 murine melanoma cells. After 48h treatment of cells with quercetin in the presence of vitamin E, L-ascorbic acid, GSH, we measured the cytotoxicities by MTT assay. The cells exhibited a dose-dependent inhibition in their proliferation in the presence of vitamin E, L-ascorbic acid, GSH respectively. We also investigated the effects of antioxidant enzyme activity and ROS generation. The antioxidant enzyme activity of quercetin in the presence of vitamin E was stronger than GSH, L-ascorbic acid, the same treatments decreased ROS generation in B16F10 murine melanoma cells. Taken together, these result demonstrate that the antioxidant effect of quercetin can enhanced in the presence of vitamin E and it might plays an important role in anti-oxidative effects.

• Journal of Preventive Medicine and Public Health
• /
• 제34권4호
• /
• pp.444-446
• /
• 2001

Objectives : Toxoplasmosis is a member of the zoonosis group and may cause congenital infection . Antibody positive rates of toxoplasmosis were examined in high school students in Cheju, Korea to facilitate the study aim of examining the seroprevalence of Toxoplasma gondii in school teachers of child-bearing age in Cheju Island. Methods : The study population comprised teachers of child-bearing age in primary, middle and high schools, aged 35 years and younger, who wished to be tested for Toxoplasma gondii antibodies (IgG) by the indirect latex agglutination test (ILA) and the enzyme-linked immunosorbent assay (ELISA) method. Results : The overall antibody positive rate was 3.8% in the study subjects (n=314), a rate which showed no significant difference due to birth place, history of bringing up pets, or history of contacting a cat. Conclusion : We confirmed that the seroprevalence of Toxoplasma gondii in a population of child-bearing teachers in Cheju Island was the same as that previously reported in other parts of Korea.