Reactive Oxygen Species in HaCaT Keratinocytes After UVB Irradiation Are Triggered by Intracellular Ca2+ Levels

Hashimoto, Masaki; Izutsu, Yukiko; Yahagi, Shoichi; Okano, Yasuaki

doi:10.1038/jidsymp.2009.12

Cited by 75 publications

(53 citation statements)

References 10 publications

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“…The peak level of ROS, registered at 2 h, is in accordance with other findings which demonstrated significantly higher rate of ROS production in the early time period after irradiation (0-2 h) (Dhumrongvaraporn and Chanvorachote, 2013;Masaki et al, 2009). The maximum level of apoptosis occurred later (at 4 h), suggesting the implication of the radiation-induced ROS in this process.…”

Section: Discussionsupporting

confidence: 79%

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Modulation of the UV-B-induced Oxidative Stress and Apoptosis in HaCaT Cell Line with <i>Calluna vulgaris</i> Extract

Virág

Brie

Burz

et al. 2015

Not Bot Hort Agrobot Cluj

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The reactive oxygen species (ROS) production due to ultraviolet B (UV-B) exposure is extremely harmful to the skin. It causes lesions of DNA, proteins and lipids and leads to cellular death. In the present study the UV-B-induced ROS and subsequent apoptosis in the human keratinocyte cell line (HaCaT) were counterbalanced by a plant extract with antioxidant capacity. Some molecules modulated by common heather (Calluna vulgaris) (CV) extract through which this may exert its photoprotective effects were also identified. The ROS were evaluated with CM-H2DCFDA assay, while apoptosis and Bax-α/Bcl-xL molecules with ELISA. The extract was standardized according to its polyphenolic content and the most important biologically active compounds, such as hyperozid, quercetin, isoquercetin, kampferol were evidenced by high-performance liquid chromatography. The UV-B induced ROS production occurred at its highest level at 2 h after the exposure of the HaCaT cells, while apoptosis later, at 4 h. The most significant changes in Bax-α and Bcl-XL proteins induced by UV-B, as well as the highest effect of the extract on apoptosis, were both registered at 4 h. The CV extract decreased concentration-and time-dependently the UV-B-induced ROS production and prevented apoptosis. These effects of CV occurred, at least to a certain extent, due to the modulation of Bax-α/Bcl-XL proteins. These findings suggest that skin cells could be protected from some of the UV-B-induced harmful effects by the administration of the CV extract, which may be further exploited as a potential photoprotective agent.

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Section: Discussionsupporting

confidence: 79%

“…Ryu et al (2010) claim that a pathway linked to leukotriene B4 receptor BLT2 and NADPH oxidase family protein Nox1 might have a crucial role. Others sustain that intracellular Ca 2+ (Masaki et al, 2009) or catalase enzyme in the skin (Heck et al, 2003) might be the trigger for UV-B-induced ROS generation.…”

Section: Discussionmentioning

confidence: 99%

Modulation of the UV-B-induced Oxidative Stress and Apoptosis in HaCaT Cell Line with <i>Calluna vulgaris</i> Extract

Virág

Brie

Burz

et al. 2015

Not Bot Hort Agrobot Cluj

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“…OH) (Blokhina et al, 2003), are generated in cells via ROS-producing enzymes, such as NADPH oxidase. There is growing evidence that plasma membrane-bound NADPH oxidase is a major source of ROS production induced by UV radiation in keratinocytes (Beak et al, 2004;Park et al, 2006) and that ROS generation requires elevation of intracellular Ca 2+ level in keratinocytes (Goldman et al, 1997;Masaki et al, 2009). ROS are highly reactive and are capable of causing oxidative DNA damage (Ehlers et al, 1999), altering membrane potential (Ehlers et al, 1999), peroxidizing proteins (Perluigi et al, 2010) and lipids (Punnonen et al, 1991), and inducing apoptosis (Clayton et al, 1974).…”

Section: Introductionmentioning

confidence: 99%

Involvement of P2Y receptors in the protective effect of ATP towards the cell damage in HaCaT cells exposed to H2O2

2011

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-It has recently been reported that activation of P2Y 1 receptor, one of the purine receptors, by extracellular nucleotides induces cytoprotection against oxidative stress. In this study, we examined the protective effect of ATP on the cell damage in human epidermal keratinocyte HaCaT cells exposed to H 2 O 2 via the P2Y receptor-mediated induction of intracellular antioxidants. The cells were damaged by exposure to H 2 O 2 in a dose-and time-dependent manner. The damage induced by 7.5 mM H 2 O 2 was blocked by pretreatment of the cells with ATP (1-10 μM). The protective effect of ATP was significantly reduced by P2Y receptor antagonists. Exogenously added ATP induced various intracellular antioxidants, including thiol-containing proteins, Cu/Zn superoxide dismutase (SOD) and thioredoxin-1, in HaCaT cells. In conclusion, it was found that ATP protected the cells from the H 2 O 2 -induced cell damages via the P2Y receptor-mediated induction of intracellular antioxidants.

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“…UVB radiation is genotoxic and causing extensive cell damage. Many studies reported that UVB radiation induces ROS and leads to cell damage in epidermal keratinocytes (Heck et al, 2003;Masaki et al, 2009). The present results demonstrated that the recombinant protein of EC-SOD had a scavenging activity reached to 30% against H 2 O 2 -induced ROS compared with 27% activity of B. hamifera extract (Piao et al, 2012) at the same concentration of 200 μg mLG 1 and 67% activity for N-acetyl cysteine (NAC).…”

Section: Discussionmentioning

confidence: 99%

Protective Effects of Highly Expressed Recombinant Human EC-SOD against ROS, UVB-Induced Apoptosis and DNA-Damage in Human Keratinocytes

Muharram¹

2015

Biotechnology

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A B S T R A C T Human extracellular superoxide dismutase (hEC-SOD) is a tetramer protein protects the extracellular space from oxidative stress by catalyzing the dismutation of biologically toxic superoxide anion into hydrogen peroxide and oxygen. Difficulty in obtaining large quantity of active recombinant (rhEC-SOD) has slowed its clinical applications although different systems have been used for its expression. In this study, transformed cells with His6-tagged rhEC-SOD were grown by fed-batch fermentation yielding a final dry weight of 16.47.6 g LG 1 as an inclusion body which comprised 43.17% of total protein. Inclusion bodies of the rhEC-SOD were solubilized, refolded and purified by Immobilized Metal Affinity (IMAC) and Gel Filtration Chromatography (GFC). With 2 L of fed-batch fermentation, 56 mg rhEC-SOD could be produced (purity 98%) with a total activity of 317.33 U. Appearance of the purified rhEC-SOD as a monomer form (26 kDa) directed the work to examine the role of the signal peptide at the N-terminal region in inducing the tetramer formation of this protein. In the predicted motif of the α-helix of the N-terminal region, amino acid substitutions of (M20D, V24D, W28A and V31D) lead to the complete disruption of the tetramer form of rhEC-SOD protein. However, no contribution could be detected with the mutations of (D12A, W16A and A22D). Partial disturbance of the tetramerization appeared with the mutations of R34A and I17D. Protective effects of rhEC-SOD against Reactive Oxygen Species (ROS), UVB-Induced apoptosis and DNA fragmentation were analyzed in human HaCaT keratinocytes. The rhEC-SOD had a scavenging activity of 30 and 14% against H 2 O 2 -induced ROS and UVB-induced ROS, respectively. Also, rhEC-SOD could reduce the UVB-induced nuclear fragmentation index from 21-9 (43%). In addition, the DNA damage and fragmentation index decreased from 1.54-1.15 in UVB irradiated upon the treatment of keratinocyte HaCaT cells with rhEC-SOD protein before UVB irradiation.

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Reactive Oxygen Species in HaCaT Keratinocytes After UVB Irradiation Are Triggered by Intracellular Ca2+ Levels

Cited by 75 publications

References 10 publications

Modulation of the UV-B-induced Oxidative Stress and Apoptosis in HaCaT Cell Line with <i>Calluna vulgaris</i> Extract

Modulation of the UV-B-induced Oxidative Stress and Apoptosis in HaCaT Cell Line with <i>Calluna vulgaris</i> Extract

Involvement of P2Y receptors in the protective effect of ATP towards the cell damage in HaCaT cells exposed to H2O2

Protective Effects of Highly Expressed Recombinant Human EC-SOD against ROS, UVB-Induced Apoptosis and DNA-Damage in Human Keratinocytes

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