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The flavonoid, fisetin, inhibits UV radiation-induced oxidative stress and the activation of NF-kappaB and MAPK signaling in human lens epithelial cells.

Yao K, Zhang L, Zhang Y, Ye P, Zhu N - Mol. Vis. (2008)

Bottom Line: The effect of fisetin on the generation of reactive oxygen species (ROS) of SRA01/04 cells was determined by flow cytometry.Treatment of SRA01/04 cells with fisetin inhibited UVB-induced cell death and the generation of ROS.Fisetin inhibited UVB-induced activation and translocation of NF-kappaB/p65, which was mediated through an inhibition of the degradation and activation of IkappaB.

View Article: PubMed Central - PubMed

Affiliation: Eye Center, Affiliated Second Hospital, College of Medicine, Zhejiang University, Hangzhou, China. xlren@zju.edu.cn

ABSTRACT

Purpose: Ultraviolet (UV) radiation-induced oxidative stress plays a significant role in the progression of cataracts. This study investigated the photoprotective effect of fisetin on UV radiation-induced oxidative stress in human lens epithelial cells and the possible molecular mechanism involved.

Methods: SRA01/04 cells exposed to different doses of ultraviolet B (UVB) were cultured with various concentrations of fisetin and subsequently monitored for cell viability by the 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. The effect of fisetin on the generation of reactive oxygen species (ROS) of SRA01/04 cells was determined by flow cytometry. Translocation of nuclear factor kappa-B (NF-kappaB) was examined by immunocytochemistry. Expression of NF-kappaB/P65, inhibiter kappa B (IkappaB), and mitogen activated protein kinase (MAPK) proteins were measured by western blot.

Results: Treatment of SRA01/04 cells with fisetin inhibited UVB-induced cell death and the generation of ROS. Fisetin inhibited UVB-induced activation and translocation of NF-kappaB/p65, which was mediated through an inhibition of the degradation and activation of IkappaB. Fisetin also inhibited UVB-induced phosphorylation of the p38 and c-Jun N-terminal kinase (JNK) proteins of the MAPK family at various time points studied.

Conclusions: The flavonoid, fisetin, could be useful in attenuation of UV radiation-induced oxidative stress and the activation of NF-kappaB and MAPK signaling in human lens epithelial cells, which suggests that fisetin has a potential protective effect against cataractogenesis.

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Related in: MedlinePlus

Effect of fisetin on the viability of UVB-exposed human lens epithelial cells. A: Cell viability of SRA01/04 to different irradiation intensities of UVB (30, 60, and 90 mJ/cm2) is shown at different incubation times of 0, 3, 6, 12, 24, and 48 h. The viability was measured by MTT assay. B: The cell viability of cells pretreated with fisetin at different concentrations (0, 5, 10, 25, 50, and 100 μg/ml) for 1 h before exposure to 30 mJ/cm2 UVB was estimated using MTT assay after being cultured for 24 h. Data are represented as the mean±SD from three independent experiments. An asterisk indicates p<0.05.
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f1: Effect of fisetin on the viability of UVB-exposed human lens epithelial cells. A: Cell viability of SRA01/04 to different irradiation intensities of UVB (30, 60, and 90 mJ/cm2) is shown at different incubation times of 0, 3, 6, 12, 24, and 48 h. The viability was measured by MTT assay. B: The cell viability of cells pretreated with fisetin at different concentrations (0, 5, 10, 25, 50, and 100 μg/ml) for 1 h before exposure to 30 mJ/cm2 UVB was estimated using MTT assay after being cultured for 24 h. Data are represented as the mean±SD from three independent experiments. An asterisk indicates p<0.05.

Mentions: As shown in Figure 1, UVB irradiation produced a progressive cytotoxic effect on cultured SRA01/04 cells in a dose-dependent manner (Figure 1A). UVB irradiation at 30 mJ/cm2 significantly reduced cell viability in a time-dependent manner. The irradiation energy of 30 mJ/cm2 was thus adopted for further studies. When fisetin, in concentrations between 5 and 100 μg/ml, was added to UVB-exposed cells for 24 h, the viability was substantially enhanced in a dose-dependent manner (Figure 1B). Pretreatment of 25 μg/ml fisetin significantly inhibited UVB-induced cell damage (p<0.05; Figure 1B).


The flavonoid, fisetin, inhibits UV radiation-induced oxidative stress and the activation of NF-kappaB and MAPK signaling in human lens epithelial cells.

Yao K, Zhang L, Zhang Y, Ye P, Zhu N - Mol. Vis. (2008)

Effect of fisetin on the viability of UVB-exposed human lens epithelial cells. A: Cell viability of SRA01/04 to different irradiation intensities of UVB (30, 60, and 90 mJ/cm2) is shown at different incubation times of 0, 3, 6, 12, 24, and 48 h. The viability was measured by MTT assay. B: The cell viability of cells pretreated with fisetin at different concentrations (0, 5, 10, 25, 50, and 100 μg/ml) for 1 h before exposure to 30 mJ/cm2 UVB was estimated using MTT assay after being cultured for 24 h. Data are represented as the mean±SD from three independent experiments. An asterisk indicates p<0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2571947&req=5

f1: Effect of fisetin on the viability of UVB-exposed human lens epithelial cells. A: Cell viability of SRA01/04 to different irradiation intensities of UVB (30, 60, and 90 mJ/cm2) is shown at different incubation times of 0, 3, 6, 12, 24, and 48 h. The viability was measured by MTT assay. B: The cell viability of cells pretreated with fisetin at different concentrations (0, 5, 10, 25, 50, and 100 μg/ml) for 1 h before exposure to 30 mJ/cm2 UVB was estimated using MTT assay after being cultured for 24 h. Data are represented as the mean±SD from three independent experiments. An asterisk indicates p<0.05.
Mentions: As shown in Figure 1, UVB irradiation produced a progressive cytotoxic effect on cultured SRA01/04 cells in a dose-dependent manner (Figure 1A). UVB irradiation at 30 mJ/cm2 significantly reduced cell viability in a time-dependent manner. The irradiation energy of 30 mJ/cm2 was thus adopted for further studies. When fisetin, in concentrations between 5 and 100 μg/ml, was added to UVB-exposed cells for 24 h, the viability was substantially enhanced in a dose-dependent manner (Figure 1B). Pretreatment of 25 μg/ml fisetin significantly inhibited UVB-induced cell damage (p<0.05; Figure 1B).

Bottom Line: The effect of fisetin on the generation of reactive oxygen species (ROS) of SRA01/04 cells was determined by flow cytometry.Treatment of SRA01/04 cells with fisetin inhibited UVB-induced cell death and the generation of ROS.Fisetin inhibited UVB-induced activation and translocation of NF-kappaB/p65, which was mediated through an inhibition of the degradation and activation of IkappaB.

View Article: PubMed Central - PubMed

Affiliation: Eye Center, Affiliated Second Hospital, College of Medicine, Zhejiang University, Hangzhou, China. xlren@zju.edu.cn

ABSTRACT

Purpose: Ultraviolet (UV) radiation-induced oxidative stress plays a significant role in the progression of cataracts. This study investigated the photoprotective effect of fisetin on UV radiation-induced oxidative stress in human lens epithelial cells and the possible molecular mechanism involved.

Methods: SRA01/04 cells exposed to different doses of ultraviolet B (UVB) were cultured with various concentrations of fisetin and subsequently monitored for cell viability by the 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. The effect of fisetin on the generation of reactive oxygen species (ROS) of SRA01/04 cells was determined by flow cytometry. Translocation of nuclear factor kappa-B (NF-kappaB) was examined by immunocytochemistry. Expression of NF-kappaB/P65, inhibiter kappa B (IkappaB), and mitogen activated protein kinase (MAPK) proteins were measured by western blot.

Results: Treatment of SRA01/04 cells with fisetin inhibited UVB-induced cell death and the generation of ROS. Fisetin inhibited UVB-induced activation and translocation of NF-kappaB/p65, which was mediated through an inhibition of the degradation and activation of IkappaB. Fisetin also inhibited UVB-induced phosphorylation of the p38 and c-Jun N-terminal kinase (JNK) proteins of the MAPK family at various time points studied.

Conclusions: The flavonoid, fisetin, could be useful in attenuation of UV radiation-induced oxidative stress and the activation of NF-kappaB and MAPK signaling in human lens epithelial cells, which suggests that fisetin has a potential protective effect against cataractogenesis.

Show MeSH
Related in: MedlinePlus