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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 UVB-induced generation of reactive oxygen species in human lens epithelial cells. A: SRA01/04 cells were pretreated with 25 μg/ml fisetin for 1 h followed by exposure to 30 mJ/cm2 UVB before incubation for 2 h and being loaded with DCFH-DA. Oxidant generation was measured by DCF flow cytometry as described in Methods. B: Representative fluorescent images of controls and UVB-irradiated cells were taken under fluorescence microscopy. Magnification, 100X.
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f2: Effect of fisetin on UVB-induced generation of reactive oxygen species in human lens epithelial cells. A: SRA01/04 cells were pretreated with 25 μg/ml fisetin for 1 h followed by exposure to 30 mJ/cm2 UVB before incubation for 2 h and being loaded with DCFH-DA. Oxidant generation was measured by DCF flow cytometry as described in Methods. B: Representative fluorescent images of controls and UVB-irradiated cells were taken under fluorescence microscopy. Magnification, 100X.

Mentions: DCF-DA was used to carry out the generation of ROS. As shown in Figure 2A, treatment with 30 mJ/cm2 UVB significantly enhanced the generation of ROS from 0.9% to 26.9%. Pretreatment with 25 μg/ml fisetin distinctly reduced the generation of ROS. The generation of ROS was also examined by DCF fluorescence. There was an expected lack of staining in the UVB-free control group (Figure 2B). SRA01/04 cells exposed to UVB alone had markedly heavy staining, indicating a marked increase in ROS generation at the single cell level. UVB-exposed cells revealed a significant decrease of DCF staining in the presence of fisetin (Figure 2B).


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 UVB-induced generation of reactive oxygen species in human lens epithelial cells. A: SRA01/04 cells were pretreated with 25 μg/ml fisetin for 1 h followed by exposure to 30 mJ/cm2 UVB before incubation for 2 h and being loaded with DCFH-DA. Oxidant generation was measured by DCF flow cytometry as described in Methods. B: Representative fluorescent images of controls and UVB-irradiated cells were taken under fluorescence microscopy. Magnification, 100X.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Effect of fisetin on UVB-induced generation of reactive oxygen species in human lens epithelial cells. A: SRA01/04 cells were pretreated with 25 μg/ml fisetin for 1 h followed by exposure to 30 mJ/cm2 UVB before incubation for 2 h and being loaded with DCFH-DA. Oxidant generation was measured by DCF flow cytometry as described in Methods. B: Representative fluorescent images of controls and UVB-irradiated cells were taken under fluorescence microscopy. Magnification, 100X.
Mentions: DCF-DA was used to carry out the generation of ROS. As shown in Figure 2A, treatment with 30 mJ/cm2 UVB significantly enhanced the generation of ROS from 0.9% to 26.9%. Pretreatment with 25 μg/ml fisetin distinctly reduced the generation of ROS. The generation of ROS was also examined by DCF fluorescence. There was an expected lack of staining in the UVB-free control group (Figure 2B). SRA01/04 cells exposed to UVB alone had markedly heavy staining, indicating a marked increase in ROS generation at the single cell level. UVB-exposed cells revealed a significant decrease of DCF staining in the presence of fisetin (Figure 2B).

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