• Bulletin of the Korean Chemical Society
• /
• v.32 no.7
• /
• pp.2187-2192
• /
• 2011

Hydrogen peroxide plays a key role as a second messenger in the normal cellular signaling but its overproduction has been implicated in various life-threatening diseases. Peroxalate chemiluminescence is the light emission from a three component reaction between peroxalate, hydrogen peroxide and fluorophores. It has proven great potential as a methodology to detect hydrogen peroxide in physiological environments because of its excellent sensitivity and specificity to hydrogen peroxide. We developed chemiluminescent micelles composed of amphiphilic polymers, peroxalate and fluorescent dyes to detect hydrogen peroxide at physiological concentrations. In this work, we studied the relationship between the chemiluminescence reactivity and stability of peroxalate by varying the substitutes on the aryl rings of peroxalate. Alkyl substitutes on the aryl ring of peroxalate increased the stability against water hydrolysis, but diminished the reactivity to hydrogen peroxide. Chemiluminescent micelles encapsulating diphenyl peroxalate showed significantly higher chemiluminescence intensity than the counterpart encapsulating dimethylphenyl or dipropylphenyl peroxalate. Diphenyl peroxalate-encapsulated micelles could detect hydrogen peroxide generated from macrophage cells stimulated by lipopolysaccharide (LPS) and image hydrogen peroxide generated during LPS-induced inflammatory responses in a mouse.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.24 no.1
• /
• pp.96-101
• /
• 2010

The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium Fermented with Sacchromyces cerevisiae (AFS) on hydrogen peroxide production within human hepatocyte HepG2 cells treated with gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. AFS (0~400 ug/mL) was treated with gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. AFS showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde in HepG2 Cells. AFS could be supposed to have the hepatoprotective effect related with hepatocytologic signaling activity against gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.6
• /
• pp.1379-1384
• /
• 2009

The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium Fermented with Lactobacillus pentosus (AFL) on hydrogen peroxide production within human hepatocyte HepG2 cells treated with gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. AFL (0~400 ug/mL) was treated with gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. AFL showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde in HepG2 Cells. AFL could be supposed to have the hepatoprotective effect related with hepatocytologic signaling activity against gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde.

• International Journal of Vascular Biomedical Engineering
• /
• v.2 no.2
• /
• pp.1-9
• /
• 2004

The history of studies in biology regarding reactive oxygen species (ROS) is approximately 40 years. During the initial 30 years, it appeared that these studies were mainly focused on the toxicity of ROS. However, recent studies have identified another action regarding oxidative signaling, other than toxicity of ROS. Basically, it is suggested that ROS are reactive, and degenerate to biomolecules such as DNA and proteins, leading to deterioration of cellular functions as an oxidative stress. On the other hand, recent studies have shown that ROS act as oxidative signaling in cells, resulting in various gene expressions. Recently ROS emerged as critical signaling molecules in cardiovascular research. Several studies over the past decade have shown that physiological effects of vasoactive factors are mediated by these reactive species and, conversely, that altered redox mechanisms are implicated in the occurrence of metabolic and cardiovascular diseases ROS is a collective term often used by scientist to include not only the oxygen radicals($O2^{-{\cdot}},\;{^{\cdot}}OH$), but also some non-radical derivatives of oxygen. These include hydrogen peroxide, hypochlorous acid (HOCl) and ozone (O3). The superoxide anion ($O2^{-{\cdot}}$) is formed by the univalent reduction of triplet-state molecular oxygen ($^3O_2$). Superoxide dismutase (SOD)s convert superoxide enzymically into hydrogen peroxide. In biological tissues superoxide can also be converted nonenzymically into the nonradical species hydrogen peroxide and singlet oxygen ($^1O_2$). In the presence of reduced transition metals (e.g., ferrous or cuprous ions), hydrogen peroxide can be converted into the highly reactive hydroxyl radical (${^{\cdot}}OH$). Alternatively, hydrogen peroxide may be converted into water by the enzymes catalase or glutathione peroxidase. In the glutathione peroxidase reaction glutathione is oxidized to glutathione disulfide, which can be converted back to glutathione by glutathione reductase in an NADPH-consuming process.

• Nutrition Research and Practice
• /
• v.4 no.5
• /
• pp.351-355
• /
• 2010

Our previous proteomic study demonstrated that oxidative stress and antioxidant delphinidin regulated the cellular level of $p27^{kip1}$ (referred to as p27) as well as some heat shock proteins in human colon cancer HT 29 cells. Current study was conducted to validate and confirm the regulation of these proteins using both in vitro and in vivo systems. The level of p27 was decreased by hydrogen peroxide in a dose-dependent manner in human colon carcinoma HCT 116 (p53-positive) cells while it was increased upon exposure to hydrogen peroxide in HT 29 (p53-negative) cells. However, high concentration of hydrogen peroxide (100 ${\mu}M)$ downregulated p27 in both cell lines, but delphindin, one of antioxidative anthocyanins, enhanced the level of p27 suppressed by 100 ${\mu}M$ hydrogen peroxide. ICR mice were injected with varying concentrations of hydrogen peroxide, delphinidin and both. Western blot analysis for the mouse large intestinal tissue showed that the expression of p27 was upregulated by 25 mg/kg BW hydrogen peroxide. To investigate the association of p27 regulation with hypoxia-inducible factor 1-beta (HIF-$1{\beta}$), the level of p27 was analyzed in wild-type mouse hepatoma hepa1c1c7 and Aryl Hydrocarbon Nuclear Translocator (arnt, HIF-$1{\beta}$)-defective mutant BPRc1 cells in the absence and presence of hydrogen peroxide and delphinidin. While the level of p27 was responsive to hydrogen peroxide and delphinidin, it remained unchanged in BPRc1, suggesting that the regulation of p27 requires functional HIF-$1{\beta}$. We also found that hydrogen peroxide and delphinidin affected PI3K/Akt/mTOR signaling pathway which is one of upstream regulators of HIFs. In conclusion, hydrogen peroxide and antioxidant delphinidin seem to regulate intracellular level of p27 through regulating HIF-1 level which is, in turn, governed by its upstream regulators comprising of PI3K/Akt/mTOR signaling pathway. The results should also encourage further study for the potential of p27 as a biomarker for intracellular oxidative or antioxidant status.

• The Plant Pathology Journal
• /
• v.29 no.4
• /
• pp.427-434
• /
• 2013

2R,3R-Butanediol, a volatile compound produced by certain rhizobacteria, is involved in induced drought tolerance in Arabidopsis thaliana through mechanisms involving stomatal closure. In this study, we examined the involvement of nitric oxide and hydrogen peroxide in induced drought tolerance, because these are signaling agents in drought stress responses mediated by abscisic acid (ABA). Fluorescence-based assays showed that systemic nitric oxide and hydrogen peroxide production was induced by 2R,3R-butanediol and correlated with expression of genes encoding nitrate reductase and nitric oxide synthase. Co-treatment of 2R,3R-butanediol with an inhibitor of nitrate reductase or an inhibitor of nitric oxide synthase lowered nitric oxide production and lessened induced drought tolerance. Increases in hydrogen peroxide were negated by co-treatment of 2R,3R-butanediol with inhibitors of NADPH oxidase, or peroxidase. These findings support the volatile 2R,3R-butanediol synthesized by certain rhizobacteria is an active player in induction of drought tolerance through mechanisms involving nitric oxide and hydrogen peroxide production.

• BMB Reports
• /
• v.46 no.12
• /
• pp.600-605
• /
• 2013

Interplay between calcium ions ($Ca^{2+}$) and reactive oxygen species (ROS) delicately controls diverse pathophysiological functions of vascular smooth muscle cells (VSMCs). However, details of the $Ca^{2+}$ and ROS signaling network have been hindered by the absence of a method for dual measurement of $Ca^{2+}$ and ROS. Here, a real-time monitoring system for $Ca^{2+}$ and ROS was established using a genetically encoded hydrogen peroxide indicator, HyPer, and a ratiometric $Ca^{2+}$ indicator, fura-2. For the simultaneous detection of fura-2 and HyPer signals, 540 nm emission filter and 500 nm~ dichroic beamsplitter were combined with conventional exciters. The wide excitation spectrum of HyPer resulted in marginal cross-contamination with fura-2 signal. However, physiological $Ca^{2+}$ transient and hydrogen peroxide were practically measurable in HyPer-expressing, fura-2-loaded VSMCs. Indeed, distinct $Ca^{2+}$ and ROS signals could be successfully detected in serotonin-stimulated VSMCs. The system established in this study is applicable to studies of crosstalk between $Ca^{2+}$ and ROS.

• Asian Pacific Journal of Cancer Prevention
• /
• v.17 no.3
• /
• pp.957-963
• /
• 2016

Cellular senescence, a barrier to tumorigenesis, controls aberrant proliferation of cells. We here aimed to investigate cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines using five different inducing agents: 5-aza-2'deoxycytidine, bromodeoxyuridine, interferons ($IFN{\beta}$ and $IFN{\gamma}$), and hydrogen peroxide. We analyzed senescence characteristics, colony formation ability, expression of genes involved in cell cycling and interferon signaling pathways, and protein levels. Treatment with all five agents decreased cell proliferation and induced cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines with different degrees of growth-inhibitory effects depending on cell type and origin. Bromodeoxyuridine gave the strongest stimulus to inhibit growth and induce senescence in most cell lines tested. Expression of p21 and interferon related genes was upregulated in most conditions. The fact that bromodeoxyuridine had the strongest effects on growth inhibition and senescence induction implies that senescence in cholangiocarcinoma cells is likely controlled by DNA damage response pathways relating to the p53/p21 signaling. In addition, interferon signaling pathways may partly regulate this mechanism in cholangiocarcinoma cells.

• Biomolecules & Therapeutics
• /
• v.26 no.6
• /
• pp.584-590
• /
• 2018

Osteoporosis development is closely associated with oxidative stress and reactive oxygen species (ROS). Taurine has potential antioxidant effects, but its role in osteoblasts is not clearly understood. The aim of this study was to determine the protective effects and mechanisms of actions of taurine on hydrogen peroxide ($H_2O_2$)-induced oxidative stress in osteoblast cells. UMR-106 cells were treated with taurine prior to $H_2O_2$ exposure. After treatment, cell viability, apoptosis, intracellular ROS production, malondialdehyde content, and alkaline phosphate (ALP) activity were measured. We also investigated the protein levels of ${\beta}-catenin$, ERK, CHOP and NF-E2-related factor 2 (Nrf2) along with the mRNA levels of Nrf2 downstream antioxidants. The results showed that pretreatment of taurine could reverse the inhibition of cell viability and suppress the induced apoptosis in a dose-dependent manner: taurine significantly reduced $H_2O_2$-induced oxidative damage and expression of CHOP, while it induced protein expression of Nrf2 and ${\beta}-catenin$ and activated ERK phosphorylation. DKK1, a Wnt/${\beta}-catenin$ signaling inhibitor, significantly suppressed the taurine-induced Nrf2 signaling pathway and increased CHOP. Activation of ERK signaling mediated by taurine in the presence of $H_2O_2$ was significantly inhibited by DKK1. These data demonstrated that taurine protects osteoblast cells against oxidative damage via Wnt/${\beta}-catenin$-mediated activation of the ERK signaling pathway.

• Proceedings of the Korea Environmental Mutagen Society Conference
• /
• 2004.11a
• /
• pp.115-115
• /
• 2004