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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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Effect of fisetin on UVB-induced phosphorylation of the MAPK pathway. SRA01/04 cells were exposed to UVB (30 mJ/ cm2) with or without pretreatment with fisetin (25 μg/ml) for 1 h. Cells were harvested at different time points (30, 60, and 120 min) after UVB exposure, and cell lysates were prepared. The phosphorylated and total protein levels of ERK1/2, JNK, and p38 were detected with specific antibodies using western blot analysis as described in Methods. A representative blot from three independent experiments with identical observations is shown.
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f4: Effect of fisetin on UVB-induced phosphorylation of the MAPK pathway. SRA01/04 cells were exposed to UVB (30 mJ/ cm2) with or without pretreatment with fisetin (25 μg/ml) for 1 h. Cells were harvested at different time points (30, 60, and 120 min) after UVB exposure, and cell lysates were prepared. The phosphorylated and total protein levels of ERK1/2, JNK, and p38 were detected with specific antibodies using western blot analysis as described in Methods. A representative blot from three independent experiments with identical observations is shown.

Mentions: Data on the kinetics of MAPK activation in UVB-irradiated cells showed that phosphorylation of p38 started 30 min after irradiation and that maximum phosphorylation occurred 2 h after irradiation (Figure 4). Western blot and subsequent measurement of the intensity of the bands relative to the total amount of p38 phosphorylation indicated that treatment with fisetin markedly inhibited UVB-induced phosphorylation of p38 at each time point studied. A marked induction in JNK phosphorylation started after 30 min and remained at a high level until 2 h had passed (Figure 4). Treatment with fisetin inhibited UVB-induced phosphorylation of JNK at each time point studied. In contrast, exposure of SRA01/04 cells to UVB reduced phosphorylation of ERK1/2 (p42/p44) after 30 min, and the reduced levels of ERK1/2 phosphorylation were observed until 2 h after irradiation (Figure 4). Treatment with fisetin elevated the phosphorylation of ERK1/2 at each time point studied. Importantly, treatment of SRA01/04 cells with fisetin alone did not induce the phosphorylation of the ERK1/2, JNK, or p38 proteins of the MAPK family (data not shown). Further, the total amount of ERK1/2, JNK, and p38 remained unchanged at each time point studied.


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 phosphorylation of the MAPK pathway. SRA01/04 cells were exposed to UVB (30 mJ/ cm2) with or without pretreatment with fisetin (25 μg/ml) for 1 h. Cells were harvested at different time points (30, 60, and 120 min) after UVB exposure, and cell lysates were prepared. The phosphorylated and total protein levels of ERK1/2, JNK, and p38 were detected with specific antibodies using western blot analysis as described in Methods. A representative blot from three independent experiments with identical observations is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Effect of fisetin on UVB-induced phosphorylation of the MAPK pathway. SRA01/04 cells were exposed to UVB (30 mJ/ cm2) with or without pretreatment with fisetin (25 μg/ml) for 1 h. Cells were harvested at different time points (30, 60, and 120 min) after UVB exposure, and cell lysates were prepared. The phosphorylated and total protein levels of ERK1/2, JNK, and p38 were detected with specific antibodies using western blot analysis as described in Methods. A representative blot from three independent experiments with identical observations is shown.
Mentions: Data on the kinetics of MAPK activation in UVB-irradiated cells showed that phosphorylation of p38 started 30 min after irradiation and that maximum phosphorylation occurred 2 h after irradiation (Figure 4). Western blot and subsequent measurement of the intensity of the bands relative to the total amount of p38 phosphorylation indicated that treatment with fisetin markedly inhibited UVB-induced phosphorylation of p38 at each time point studied. A marked induction in JNK phosphorylation started after 30 min and remained at a high level until 2 h had passed (Figure 4). Treatment with fisetin inhibited UVB-induced phosphorylation of JNK at each time point studied. In contrast, exposure of SRA01/04 cells to UVB reduced phosphorylation of ERK1/2 (p42/p44) after 30 min, and the reduced levels of ERK1/2 phosphorylation were observed until 2 h after irradiation (Figure 4). Treatment with fisetin elevated the phosphorylation of ERK1/2 at each time point studied. Importantly, treatment of SRA01/04 cells with fisetin alone did not induce the phosphorylation of the ERK1/2, JNK, or p38 proteins of the MAPK family (data not shown). Further, the total amount of ERK1/2, JNK, and p38 remained unchanged at each time point studied.

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