• Journal of Nutrition and Health
• /
• v.32 no.3
• /
• pp.318-326
• /
• 1999

Apoptotic cell death, characterized by DNA fragmentation and morphological changes, has previously been shown to occur in vascular endothelial cells cultured with hydrogen peroxide. The present study examined the induction of apoptosis by hydrogen peroxide and whether pyruvate, a key glycolytic intermediate and $\alpha$-keto-monocarboxylate, can inhibit the apoptotic effects in bovine pulmonary artery endothelial cells(BPAECs). Culture with 500uM hydrogen peroxide resulted in 30% cell death and induced morphological changes and DNA fragmentation. Cell injury was inhibited by the treatment with pyruvate. Pyruvate(0.1-5.0mM), and cell viability increased in a dose-dependent manner. In the presence of pyruvate(10~20mM), the viability was improved to over 95%. In contrast, treatment with lactate, a reduced form of phyuvate, did not protect against cell death oxidative stress-induced loss of viability and apoptosis was examined with $\alpha$-cyano-3-hydroxycinnarmate(COHC) as a selective mitochondrial monocarboxylate transport blocker. Incubation with COHC(500uM) did not significantly affect cell viability in the presence of hydrogen peroxide. The cytoprotection by pyruvate(3mM)against hydrogen peroxide stress was abolished by COHC. This indicates that the cytoprotection by pyruvate against oxidative stress in endothelial cells is mediated, at least in part, by mitochondrial pyruvate uptake and hence endothelial enerygetics. However, cytosolic mechanisms related, at least in part, by mitochondrial pyruvate uptake and hence endothelial energetics. However, cytosolic mechanisms related to the glutathione system may also contribute. The results suggest that pyruvate has therapeutic potential in the treatment of oxidative stress-induced cytotoxicity associated with increased apoptosis.

• Toxicological Research
• /
• v.16 no.3
• /
• pp.179-185
• /
• 2000

Zinc is known to generate reactive oxygen species (ROS) including superoxide anion and hydrogen peroxide ($H_2O_2$), which eventually contribute to cytotoxicity in a variety of cell types. Here in, we demonstrated that zinc decreased the viability of C6 glial cells in a time and dose-dependent manner, which was revealed as apoptosis characterized by ladder-pattern fragmentation of genomic DNA. chromatin condensation and DNA fragmentation in Hoechst dye staining. Zinc-induced apoptosis of C6 glial cells was prevented by the addition of catalase and antioxidants including reduced glutathione (GSH), N-acetyl-L-cysteine (NAC) and pyrrolidinedithiocarbamate (PDTC). Wefurther confirmed that zinc decreased intrac-ellular levels of GSH and generated $H_2O_2$in C6 glial cells. Moreover, antioxidants also decreased the generation of zinc-induced $H_2O_2$ in C6 glial cells. These data indicated that zinc-induced the apoptotic death of C6 glial cells via generation of reactive oxygen species such as $H_2O_2$.

• Biomolecules & Therapeutics
• /
• v.20 no.5
• /
• pp.487-491
• /
• 2012

The aim of this study was to investigate the inhibitory effects of rutaecarpine on DNA strand breaks and apoptosis induced by hydrogen peroxide ($H_2O_2$) in murine Hepa-1c1c7 cells. Oxidative DNA damage was estimated by nuclear condensation assessment, fluorescence-activated cell sorting analysis, and Comet assay. Rutaecarpine inhibited cell death induced by $500{\mu}M$ $H_2O_2$, as assessed by 4',6-diamidino-2-phenylindole (DAPI) staining. Treatment with rutaecarpine reduced the number of DNA strand breaks induced by $H_2O_2$, as assessed by DAPI staining and Comet assay, and increased quinone reductase, phosphatidylinositol 3-kinase, and pAkt protein levels, as assessed by western blotting.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.10
• /
• pp.1877-1884
• /
• 2017

Mesenchymal stem cells (MSCs) have been suggested as a primary candidate for cell therapy applications because they have self-renewal and differentiation capabilities. Although they can be expanded in ex vivo system, clinical application of these cells is still limited because they survive poorly and undergo senescence or apoptosis when transplanted and exposed to environmental factors such as oxidative stress. Thus, reducing oxidative stress is expected to improve the efficacy of MSC therapy. The milk protein lactoferrin is a multifunctional iron-binding glycoprotein that plays various roles, including reduction of oxidative stress. Thus, we explored the effect of lactoferrin on oxidative stress-induced senescence and apoptosis of human MSCs (hMSCs). Measurement of reactive oxygen species (ROS) revealed that lactoferrin inhibited the production of hydrogen peroxide-induced intracellular ROS, suggesting lactoferrin as a good candidate as an antioxidant in hMSCs. Pretreatment of lactoferrin suppressed hydrogen peroxide-induced senescence of hMSCs. In addition, lactoferrin reduced hydrogen peroxide-induced apoptosis via inhibition of caspase-3 and Akt activation. These results demonstrate that lactoferrin can be a promising factor to protect hMSCs from oxidative stress-induced senescence and apoptosis, thus increasing the efficacy of MSC therapy.

• Biomedical Science Letters
• /
• v.28 no.4
• /
• pp.237-246
• /
• 2022

Ischemia-reperfusion results in excess reactive oxygen species (ROS) that affect myocardial cell damage. ROS production inhibition is effectively proposed in treating cardiovascular diseases including myocardial hypertrophy. Studies have shown that oxidizing cultured cells in in vitro experiments gradually decreases the permeability of mitochondrial membranes time- and concentration-dependent, resulting in increased mitochondrial membrane damage due to secondary ROS production and cardiolipin loss. However, recent studies have shown that 5-iodo-6-amino-1,2-benzopyrone (INH₂BP), an anticancer and antiviral drug, inhibited peroxynitrite-induced cell damage in in vitro and alleviated partial or overall inflammation in animal experiments. Therefore, in this paper, we studied the preventive effect of INH₂BP on H9c2 cells derived from mouse heart damaged by oxidative stress using 700 μM of hydrogen peroxide. As a result of oxidative stress to H9c2 cells by hydrogen peroxide whether the treatment of INH₂BP or not, hydrogen peroxide caused serious damage in H9c2 cells. These results were confirmed with cell viability and Hoechst 33342 assays. And this damage was through cell death. However, it was confirmed that H9c2 cells pretreated with INH₂BP significantly reduced cell death by hydrogen peroxide. In addition, measurements with DCF-DA assay to determine whether ROS is produced in H9c2 cells treated with only hydrogen peroxide produced ROS significantly, but H9c2 cells pretreated with INH₂BP significantly reduced ROS production by hydrogen peroxide. Taken together, it is believed that INH₂BP can be useful for the prevention and treatment of cardiovascular diseases induced through oxidative stress such as heart damage caused by ischemia/reperfusion.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.2
• /
• pp.404-407
• /
• 2007

This study was designed to investigate the protective effect of Bojungbangam-tang Ethanol Extracts (EBJT) on cisplatin and hydrogen peroxide-induced cytotoxicity of human endothelial cell line ECV304 cells. After cells were treated with cisplatin and hydrogen peroxide, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, we used the several measures of apoptosis to determine whether this processes was involved in cisplatin and hydrogen peroxide-induced cell damage in ECV304 cells. Also, cells were treated with EBJT and then, followed by the addition of cisplatin or hydrogen peroxide. Cisplatin or hydrogen peroxide decreased the viability of ECV304 cells in a dose-dependent manner. ECV304 cells treated cisplatin or hydrogen peroxide were revealed as apoptosis characterized by nuclear staining. EBJT protected ECV304 cells from cisplatin or hydrogen peroxide-induced nuclear fragmentation and chromatin condensation. Also, EBJT inhibited the cleavage of poly(ADP-ribose) polymerase (PARP) in cisplatin or hydrogen peroxide-treated ECV304 cells. According to above results, EBJT may protect ECV304 cells from the apotosis induced by cisplatin or hydrogen peroxide.

• BMB Reports
• /
• v.39 no.2
• /
• pp.145-149
• /
• 2006

To analyze the effect of Maltol on the apoptosis of Human Neuroblastoma Cells (SH-SY5Y) treated by free radical which was generated from Hydrogen Peroxide ($H_2O_2$), flow cytometry analysis on Phosphatidylserine (PS) inverting percentage was applied to determine the apoptosis. MTT (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay was employed to analyze the cell viability. DNA electrophoresis was used to detect DNA fragmentation. Moreover intracellular calcium of concentration ($[Ca^{2+}]_i$) was measured by fluorescence emission. Flow cytometry analysis on the function of mitochondria and Western blto analysis of NF-${\kappa}B$. The results showed that the pretreatment with maltol for 2 hours could prevent the $H_2O_2$-induced apoptosis. Maltol could reduce the inverting percentage of PS, DNA fragmentation and $[Ca^{2+}]_i$, and enhance the cellular function of mitochondria. NF-${\kappa}B$ activated by $H_2O_2$ is reduced. The experiments suggest that maltol could effectively inhibit the apoptosis induced by $H_2O_2$. As a novel anti-oxidant, maltol is a new promising drug in protecting the neurological cells from the damage by free radical.

• Biomolecules & Therapeutics
• /
• v.21 no.4
• /
• pp.270-276
• /
• 2013

Fucoxanthin is an important carotenoid derived from edible brown seaweeds and is used in indigenous herbal medicines. The aim of the present study was to examine the cytoprotective effects of fucoxanthin against hydrogen peroxide-induced cell damage. Fucoxanthin decreased the level of intracellular reactive oxygen species, as assessed by fluorescence spectrometry performed after staining cultured human HaCaT keratinocytes with 2',7'-dichlorodihydrofluorescein diacetate. In addition, electron spin resonance spectrometry showed that fucoxanthin scavenged hydroxyl radical generated by the Fenton reaction in a cell-free system. Fucoxanthin also inhibited comet tail formation and phospho-histone H2A.X expression, suggesting that it prevents hydrogen peroxide-induced cellular DNA damage. Furthermore, the compound reduced the number of apoptotic bodies stained with Hoechst 33342, indicating that it protected keratinocytes against hydrogen peroxide-induced apoptotic cell death. Finally, fucoxanthin prevented the loss of mitochondrial membrane potential. These protective actions were accompanied by the down-regulation of apoptosis-promoting mediators (i.e., B-cell lymphoma-2-associated ${\times}$ protein, caspase-9, and caspase-3) and the up-regulation of an apoptosis inhibitor (B-cell lymphoma-2). Taken together, the results of this study suggest that fucoxanthin defends keratinocytes against oxidative damage by scavenging ROS and inhibiting apoptosis.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.3
• /
• pp.774-782
• /
• 2004

The water extract of Dancheonhwan (DCH) has been used to treat ischemic brain and heart damage in oriental medicine. However, little is known about the mechanism by which the water extract of DCH rescues cells from ischemic damage. Therefore, this study was designed to investigate the protective mechanisms of DCH on the H₂O₂-induced toxicity in H9c2 cardiomyoblast cells. Treatment of H₂O₂ markedly decreased the viability of H9c2 cardiomyoblast in a dose-dependent and time-dependent manner. The nature of H₂O₂-induced toxicity of H9c2 cells resulted from apoptotic death confirmed with genomic DNA fragmentation. DCH increased the viability of H₂O₂-treated H9c2 cells by about 23%, and partially suppressed the genomic DNA fragmentation and PARP cleavage. H₂O₂ also activated caspase-3 protease and -9 protease, but not both caspase-6 protease and -8 protease. H₂O₂ induced the mitochondria dysfunction, including mitochondria membrane permeability transition (MPT) and cytosolic release of cytochrome c from mitochondria, which was prevented in part by pretreatment of DCH. N-acetylcystein (NAC), a free-radical scavenger, alone increased the viability of H₂O₂-treated H9c2 cells in a dose-dependent manner. Furthermore, the combination of NAC with DCH significantly increased the viability of the H₂O₂-treated H9c2 cells in a dose-dependent manner. These data indicate that DCH has the protective effect on ROS-induced apoptosis of cadiomyoblast H9c2 cells.

• Journal of Society of Preventive Korean Medicine
• /
• v.19 no.3
• /
• pp.141-154
• /
• 2015

Objective : In this paper, we investigated the anti-oxidative capacities and protective effects of water extract of palmul-tang (PMT) against Ultraviolet B(UVB)-induced oxidative damage in human keratinocytes(HaCaT). Method : To evaluate the anti-oxidative activities of PMT, we measured scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation and reducing power of PMT. To give an oxidative stress to HaCaT cells, UVB was irradiated with $40mJ/cm^2$ to HaCaT cells. To detect the protective effects of PMT against UVB, we measured cell viability, apoptotic bodies and reactive oxygen species(ROS) generation. Results : PMT showed the anti-oxidative activities by scavenging DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation. Also PMT showed high reducing values. The UVB-induced oxidative conditions led to the cell apoptosis. However, treatment with PMT reduced oxidative stress conditions, including inhibition of cell apoptosis and expression of ROS. Conclusion : PMT had anti-oxidative activities and exhibited protective effects against UVB on HaCaT cells. PMT would be useful for the development of cosmetics treating UVB-induced skin aging.