• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.10a
• /
• pp.103-103
• /
• 2018

Oenothera biennis, commonly known as evening primrose, a potential source of natural bioactive substances: flavonoids, steroids, tannins, fatty acids and terpenoids responsible for a diverse range of pharmacological functions. However, whether extract prepared from aerial part of O. biennis (APOB) protects skin against oxidative stress remains unknown. To investigate the protective effects of APOB against oxidative stress-induced cellular damage and elucidated the underlying mechanisms in the HaCaT human skin keratinocytes. Our results revealed that treatment with APOB prior to hydrogen peroxide ($H_2O_2$) exposure significantly increased viability, and the highest DPPH radical-scavenging activities and reducing power of HaCaT cells. APOB also effectively attenuated H2O2-induced comet tail formation and inhibited the $H_2O_2$-induced phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V-positive cells. In addition, APOB exhibited scavenging activity against intracellular reactive oxygen species (ROS) accumulation and restored the mitochondrial membrane potential loss by $H_2O_2$. Moreover, $H_2O_2$ enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase (PARP), a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with APOB. Furthermore, APOB increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, APOB is able to protect HaCaT cells from $H_2O_2$-induced DNA damage and cell death through blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating the Nri2/HO-1 signaling pathway.

• Molecular & Cellular Toxicology
• /
• v.3 no.2
• /
• pp.98-106
• /
• 2007

Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. In this study, we investigated to examine gene expression profiles and genotoxic response in TK6 cells treated with DNA damaging agents MNNG (N-methyl-N'-nitrosoguanidine) and hydrogen peroxide $(H_2O_2)$. We extracted total RNA in three independent experiments and hybridized cRNA probes with oligo DNA chip (Applied Biosystems Human Genome Survey Microarray). We analyzed raw signal data with R program and AVADIS software and identified a number of deregulated genes with more than 1.5 log-scale fold change and statistical significancy. We indentified 14 genes including G protein alpha 12 showing deregulation by MNNG. The deregulated genes by MNNG represent the biological pathway regarding MAP kinase signaling pathway. Hydrogen peroxide altered 188 genes including sulfiredoxins. These results show that MNNG and $H_2O_2$ have both uniquely regulated genes that provide the potential to serve as biomarkers of exposure to DNA damaging agents.

• Applied Chemistry for Engineering
• /
• v.17 no.6
• /
• pp.638-643
• /
• 2006

Pyrolytic reaction study of dichloromethane ($CH_{2}Cl_{2}$) in excess hydrogen was performed to investigate pyrolytic reaction pathways at a pressure of 1 atm with residence times of 0.3~2.0 sec in the temperature range of $525{\sim}900^{\circ}C$. A constant feed molar ratio $CH_{2}Cl_{2}$:$H_{2}$ of 4:96 was maintained through the experiment. Reagent loss and product formation were monitored by using an on-line gas chromatograph, where batch samples were analyzed by GC/MS. Complete destruction(99%) of the parent reagent was observed at temperature near $780^{\circ}C$ with residence time over 1 sec. Major products observed were $CH_{3}Cl$, $CH_{4}$, $C_{2}H_{4}$, $C_{2}H_{6}$, and HCl. Minor products included $CHClCCl_{2}$, CHClCHCl, $CH_{2}CHCl$, and $C_{2}H_{2}$. The pyrolytic reaction pathways to describe the important features of intermediate product distributions and reagent loss, based upon thermodynamic and kinetic principles, were suggested. The results of this work provided a better understanding of pyrolytic decomposition processes which occur during the pyrolysis of $CH_{2}Cl_{2}$ and similar chlorinated methanes.

• The Korean Journal of Physiology and Pharmacology
• /
• v.24 no.1
• /
• pp.69-79
• /
• 2020

Aging is one of the risk factors for the development of cardiovascular diseases. During the progression of cellular senescence, cells enter a state of irreversible growth arrest and display resistance to apoptosis. As a flavonoid, quercetin induces apoptosis in various cells. Accordingly, we investigated the relationship between quercetin-induced apoptosis and the inhibition of cellular senescence, and determined the mechanism of oxidative stress-induced vascular smooth muscle cell (VSMC) senescence. In cultured VSMCs, hydrogen peroxide (H₂O₂) dose-dependently induced senescence, which was associated with increased numbers of senescence-associated β-galactosidase-positive cells, decreased expression of SMP30, and activation of p53-p21 and p16 pathways. Along with senescence, expression of the anti-apoptotic protein Bcl-2 was observed to increase and the levels of proteins related to the apoptosis pathway were observed to decrease. Quercetin induced apoptosis through the activation of AMP-activated protein kinase. This action led to the alleviation of oxidative stress-induced VSMC senescence. Furthermore, the inhibition of AMPK activation with compound C and siRNA inhibited apoptosis and aggravated VSMC senescence by reversing p53-p21 and p16 pathways. These results suggest that senescent VSMCs are resistant to apoptosis and quercetin-induced apoptosis attenuated the oxidative stress-induced senescence through activation of AMPK. Therefore, induction of apoptosis by polyphenols such as quercetin may be worthy of attention for its anti-aging effects.

• Korean Journal of Animal Reproduction
• /
• v.20 no.4
• /
• pp.427-432
• /
• 1997

These studies were performed to approach the precise pathway inducing the meiotic inhibitory action of hypoxanthine on mouse follicular oocytes and to identify the cause of detrimental effect of hypoxanthine on viability of the oocyte in vitro. In addition, a correlation between the meiotic inhibitory effect and the detrimental effect of hypoxanthine was investigated. Mouse follicular oocytes at germinal vesicle(GV) stage were collected from the ovaries of ICR mice by puncturing the antral follicles with a fine needle, at 48 hours after PMSG injection. Oocytes were cultured in Modified Whittingham's T6 media containing hypoxanthine and several materials that involved in metabolism of hypoxanthine, and the effects of the materials on the actions of hypoxanthine were investigated by observing germinal vesicle breake down (GVBD), 1st polar body (PB) extrusion and viability of the oocytes. Phophodiesterase significantly reduced the meiotic inhibitory effect of dbcAMP but did not influence on the inhibitory effect of hypoxanthine. Allopurinol and 6-MP significantly enhanced the meiotic inhibitory effect of hypoxanthine, but the materials themselves also showed the meiotic inhibitory action like hypoxanthine. Hypoxanthine-guanine phosphoribosyltransferase significantly enhanced the meiotic inhibitory effect of hypoxanthine, on the contrary HGPRT itself promoted meiotic resumption of the oocytes. Catalase did not induce any change in the meiotic inhibitory effect of hypoxanthine, but SOD increased the GVBD rate suppressed by hypoxanthine. The detrimental effect of hypoxanthine on viability of the oocytes was significantly reduced by allopurinol and catalase, but SOD increased the GVBD rate suppressed by hypoxanthine. The detrimental effect of hypoxanthine on viability of the oocytes was significantly reduced by allopurinol and catalase, but SOD did not reduce the deterimental effect of hypoxanthine. In conclusion, the meiotic inhibtory effect of hypoxanthine may be caused by guanyl dervartives converted from hypoxanthine via salvage pathway, and superoxide anion may partially participate in the inhibitory effect of hypoxanthine. The detrimental effect of hypoxanthine on viability of the oocytes be cused by hydrogen peroxide produced during the metabolism of hypoxanthine.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.4
• /
• pp.841-848
• /
• 2008

Archyranthes radix has had extensive therapeutic application, and there has been increasing interest in its biological effects. However, the biochemical effects of Archyranthes radix on chondrocyte oxidative stress have never been systematically investigated. Therefore, we investigated the effects of Acyranthes radix on role of MAPK signal transduction pathway on oxidative stress induced by hydrogen peroxide in rat articular chondrocytes. The statistically significant inhibitory action of Archyranthes radix on cell proliferation was observed at above $5{\mu}g/m{\ell}$. Next, we examined the time-dependent effect of $5{\mu}g/m{\ell}$ Archyranthes radix on cell proliferaion. Archyranthes radix significantly inhibited cell proliferation from 12 hr after treatment (P<0.05). $H_2O_2$, resulted in a time- and dose-dependent cell proliferation, which was largely attributed to oxidative damage. Acyranthes radix and $H_2O_2$ treatment caused marked sustained activation of phosphorylation of ERK1/2. Moreover, the synergistic phosphorylation of p44/42 MAPK by $H_2O_2$ and Archyranthes radix was selectively inhibited by PD 98059, a p44/42 MAPK inhibitor. In conclusion, these results are consistent with the hypothesis that under conditions of oxidative stress, the $H_2O_2$-induced inhibition of cell proliferation in the rat chondrocyte is mediated through a modulation of the Archyranthes radix signaling pathway, promoting further phosphorylation of p44/42 MAPK, indicating a potentially important role in cartilage repair and in the treatment of osteoarthritic cartilage.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.4
• /
• pp.888-894
• /
• 2009

It has been reported that silibinin, a natural polyphenolic flavonoid, induces cell death in various cancer cell types. However, the underlying mechanisms by which silibinin induces apoptosis in human glioma cells are poorly understood. The present study was therefore undertaken to examine the effect of silibinin on glioma cell apoptosis and to determine its underlying mechanism in human glioma cells. Apoptosis was estimated by FACS analysis. Reactive oxygen species (ROS) generation and mitochondrial membrane potential (${\Psi}m$) were measured using fluorescence dyes DCFH-DA and $DiOC_6$(3), respectively. Cytochrome c release from mitochondria and caspase-3 activation were estimated by Western blot analysis using specific antibodies. Exposure of cells to 30 mM silibinin induced apoptosis starting at 6 h, with increasing effects after 12-48h in a time-dependent manner. Silibinin caused ROS generation and disruption of ym, which were associated with the silibinin-induced apoptosis. The silibinin-induced ROS generation and disruption in ym were prevented by inhibitors of mitochondrial electron transport chain. The hydrogen peroxide scavenger catalase blocked ROS generation and apoptosis induced by silibinin. Silibinin induced cytochrome c release into cytosolic fraction and its effect was prevented by catalase and cyclosporine A. Silibinin treatment caused caspase-3 activation, which was inhibited by DVED-CHO and cyclosporine A. Pretreatment of caspase inhibitors also protected against the silibinin-induced apoptosis. These findings indicate that ROS generation plays a critical role in the initiation of the silibinin-induced apoptotic cascade by mediation of the mitochondrial apoptotic pathway including the disruption of ${\Psi}m$, cytochrome c release, and caspase-3 activation.

• BMB Reports
• /
• v.46 no.12
• /
• pp.594-599
• /
• 2013

The anti-inflammatory activity of eriodictyol and its mode of action were investigated. Eriodictyol suppressed tumor necrosis factor (mTNF)-${\alpha}$, inducible nitric oxide synthase (miNOS), interleukin (mIL)-6, macrophage inflammatory protein (mMIP)-1, and mMIP-2 cytokine release in LPS-stimulated macrophages. We found that the anti-inflammatory cascade of eriodictyol is mediated through the Toll-like Receptor (TLR)4/CD14, p38 mitogen-activated protein kinases (MAPK), extracellular-signal-regulated kinase (ERK), Jun-N terminal kinase (JNK), and cyclooxygenase (COX)-2 pathway. Fluorescence quenching and saturation-transfer difference (STD) NMR experiments showed that eriodictyol exhibits good binding affinity to JNK, $8.79{\times}10^5M^{-1}$. Based on a docking study, we propose a model of eriodictyol and JNK binding, in which eriodictyol forms 3 hydrogen bonds with the side chains of Lys55, Met111, and Asp169 in JNK, and in which the hydroxyl groups of the B ring play key roles in binding interactions with JNK. Therefore, eriodictyol may be a potent anti-inflammatory inhibitor of JNK.

• Journal of the Korean Chemical Society
• /
• v.37 no.8
• /
• pp.735-743
• /
• 1993

The electrocatalytic reduction of oxygen is investigated by cyclic voltammetry and chronoamperometry at glassy carbon electrode and carbon microelectrode coated with a variety of cobalt phenylprophyrins in various pH solutions. Oxygen reduction catalyzed by the monomeric porphyrin Co(Ⅱ)-TPP mainly occurs through the 2e$^-$ reduction pathway resulting in the formation of hydrogen peroxide whereas electrocatalytic process carried out 4e$^-$ reduction pathway of oxygen to H$_2$O at the electrodes coated with cofacial bis-cobalt phenylporphyrins in acidic solution. The electrocatalytic reduction of oxygen is irreversible and diffusion controlled. The reduction potentials of oxygen in various pH solutions have a straight line from pH 4 to pH 13, but level off in strong acidic solution. The reduction potentials of oxygen shift to positive potential more 400 mV at the electrode coated with monomer Co-TPP compound than bare glassy carbon electrode while 750 mV at the electrode coated with dimer Co-TPP compound.

• Journal of the Korean Chemical Society
• /
• v.37 no.4
• /
• pp.462-469
• /
• 1993

The electrocatalytic reduction of dioxygen is investigated by cyclic voltammetry and chronoamperometry at glassy carbon electrode and carbon microelectrode coated with a variety of cobalt phenylporphyrins. The n value obtained at carbon microelectrode is slightly different from that determined at glassy carbon electrode. Dioxygen reduction catalyzed by the monormeric porphyrin Co(II)-TPP mainly occurs through the $2e^-$ reduction pathway resulting in the formation of hydrogen peroxide, electrocatalytic process carries out $4e^-$ reduction pathway of dioxygen to $H_2O$ at the electrodes coated with bis-cobalt phenylporphyrins. The electrocatalytic reduction of dioxygen is irreversible and diffusion controlled.