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p53 activation contributes to patulin-induced nephrotoxicity via modulation of reactive oxygen species generation.

Jin H, Yin S, Song X, Zhang E, Fan L, Hu H - Sci Rep (2016)

Bottom Line: Patulin is a major mycotoxin found in fungal contaminated fruits and their derivative products.To the best of our knowledge, this is the first report addressing the functional role of p53 in patulin-induced oxidative stress.The findings of the present study provided novel insights into understanding mechanisms behind oxidative stress in response to patulin exposure.

View Article: PubMed Central - PubMed

Affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, Department of Nutrition and Health, College of Food Science and Nutritional Engineering, China Agricultural University, No17 Qinghua East Road, Haidian District, Beijing 100083, China.

ABSTRACT
Patulin is a major mycotoxin found in fungal contaminated fruits and their derivative products. Previous studies showed that patulin was able to induce increase of reactive oxygen species (ROS) generation and oxidative stress was suggested to play a pivotal role in patulin-induced multiple toxic signaling. The objective of the present study was to investigate the functional role of p53 in patulin-induced oxidative stress. Our study demonstrated that higher levels of ROS generation and DNA damage were induced in wild-type p53 cell lines than that found in either knockdown or knockout p53 cell lines in response to patulin exposure, suggesting p53 activation contributed to patulin-induced ROS generation. Mechanistically, we revealed that the pro-oxidant role of p53 in response to patulin was attributed to its ability to suppress catalase activity through up-regulation of PIG3. Moreover, these in vitro findings were further validated in the p53 wild-type/knockout mouse model. To the best of our knowledge, this is the first report addressing the functional role of p53 in patulin-induced oxidative stress. The findings of the present study provided novel insights into understanding mechanisms behind oxidative stress in response to patulin exposure.

No MeSH data available.


Related in: MedlinePlus

p53 activation promoted patulin-induced oxidative stress in vivo.(A). ROS levels were measured in p53wide type/knockout mice in response to patulin exposure. Frozen kidney sections were stained with 10 μM H2DCFDA for 30 min at 37 °C. Cells staining positively for the oxidized dye were identified by confocal microscopy. (B-D). Oxidative injury caused by patulin in p53 wide type/knockout mice. Kidney damage was assessed by measuring GSH (B), CAT (C) and LPO (D). (E). Western blotting analysis of p53, PIG3 and H2AX phosphorylation in kidney tissues. **P < 0.01. (The blots shown are representative of three independent experiments).
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f6: p53 activation promoted patulin-induced oxidative stress in vivo.(A). ROS levels were measured in p53wide type/knockout mice in response to patulin exposure. Frozen kidney sections were stained with 10 μM H2DCFDA for 30 min at 37 °C. Cells staining positively for the oxidized dye were identified by confocal microscopy. (B-D). Oxidative injury caused by patulin in p53 wide type/knockout mice. Kidney damage was assessed by measuring GSH (B), CAT (C) and LPO (D). (E). Western blotting analysis of p53, PIG3 and H2AX phosphorylation in kidney tissues. **P < 0.01. (The blots shown are representative of three independent experiments).

Mentions: The above data have established a pro-oxidant function of p53 in response to patulin exposure in the cell culture model. We further validated these in vitro findings in a homozygous p53 knockout mouse model. To know the kinetic process of patulin-induced oxidative stress in vivo, we carried out a time-course experiment using p53 wild-type mouse model. Patulin was given by i.p injection for 1, 3, 6 or 12 h and then the samples were collected for analysis of the oxidative stress biomarkers. As shown in Fig. S1A, patulin caused a rapid (as early as 3 h) inhibition of glutathione and catalase activity and increase of lipid oxidation. These effects were gradually decreased starting from 12 h of exposure. Based on the time-course results, 3 h exposure design was chosen to investigate the functional role of p53 in patulin-induced oxidative stress in vivo. As shown in Fig. 6A, Fluorescence microscope observation showed that patulin caused a significant elevated DCF fluorescence intensity in the kidney tissues of p53 WT mice. In contrast, the DCF fluorescence intensity by patulin was dramatically attenuated in the kidney tissues of p53 knockout mice, supporting p53-dependent ROS generation by patulin in vivo. Consistent with the ROS data, the contents of glutathione (GSH) (Fig. 6B) and catalase activity (Fig. 6C) were significantly decreased in response to patulin exposure, accompanied by increased lipid oxidation (LPO) (Fig. 6D), in p53 WT mice, whereas such changes were not significant in p53 KO mice at the experimental condition. Accordingly, the induction of p53 and its target PIG3 was observed in patulin-treated p53 WT mice, which was paralleled with the increased H2AX phosphorylation. PIG3 was not detectable in p53 KO mice, which was probably due to p53 deficiency, whereas a slightly increased H2AX phosphorylation in response to patulin exposure was found in p53 KO mice (Fig. 6E). We also analyzed the histopathological changes by patulin in both p53 wild-type and knockout mice and the results are shown in Fig. S1B. No obvious pathological changes were found in the kidney tissues of both p53 wild-type (1, 3, 6 or 12 h treatment) and knockout mice (3 h treatment), suggesting continuous treatment and persistent oxidative stress may be required for a detectable pathological damage. Taken together, these data clearly supported a pro-oxidant role of p53 in response to patulin in the animal model.


p53 activation contributes to patulin-induced nephrotoxicity via modulation of reactive oxygen species generation.

Jin H, Yin S, Song X, Zhang E, Fan L, Hu H - Sci Rep (2016)

p53 activation promoted patulin-induced oxidative stress in vivo.(A). ROS levels were measured in p53wide type/knockout mice in response to patulin exposure. Frozen kidney sections were stained with 10 μM H2DCFDA for 30 min at 37 °C. Cells staining positively for the oxidized dye were identified by confocal microscopy. (B-D). Oxidative injury caused by patulin in p53 wide type/knockout mice. Kidney damage was assessed by measuring GSH (B), CAT (C) and LPO (D). (E). Western blotting analysis of p53, PIG3 and H2AX phosphorylation in kidney tissues. **P < 0.01. (The blots shown are representative of three independent experiments).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4829895&req=5

f6: p53 activation promoted patulin-induced oxidative stress in vivo.(A). ROS levels were measured in p53wide type/knockout mice in response to patulin exposure. Frozen kidney sections were stained with 10 μM H2DCFDA for 30 min at 37 °C. Cells staining positively for the oxidized dye were identified by confocal microscopy. (B-D). Oxidative injury caused by patulin in p53 wide type/knockout mice. Kidney damage was assessed by measuring GSH (B), CAT (C) and LPO (D). (E). Western blotting analysis of p53, PIG3 and H2AX phosphorylation in kidney tissues. **P < 0.01. (The blots shown are representative of three independent experiments).
Mentions: The above data have established a pro-oxidant function of p53 in response to patulin exposure in the cell culture model. We further validated these in vitro findings in a homozygous p53 knockout mouse model. To know the kinetic process of patulin-induced oxidative stress in vivo, we carried out a time-course experiment using p53 wild-type mouse model. Patulin was given by i.p injection for 1, 3, 6 or 12 h and then the samples were collected for analysis of the oxidative stress biomarkers. As shown in Fig. S1A, patulin caused a rapid (as early as 3 h) inhibition of glutathione and catalase activity and increase of lipid oxidation. These effects were gradually decreased starting from 12 h of exposure. Based on the time-course results, 3 h exposure design was chosen to investigate the functional role of p53 in patulin-induced oxidative stress in vivo. As shown in Fig. 6A, Fluorescence microscope observation showed that patulin caused a significant elevated DCF fluorescence intensity in the kidney tissues of p53 WT mice. In contrast, the DCF fluorescence intensity by patulin was dramatically attenuated in the kidney tissues of p53 knockout mice, supporting p53-dependent ROS generation by patulin in vivo. Consistent with the ROS data, the contents of glutathione (GSH) (Fig. 6B) and catalase activity (Fig. 6C) were significantly decreased in response to patulin exposure, accompanied by increased lipid oxidation (LPO) (Fig. 6D), in p53 WT mice, whereas such changes were not significant in p53 KO mice at the experimental condition. Accordingly, the induction of p53 and its target PIG3 was observed in patulin-treated p53 WT mice, which was paralleled with the increased H2AX phosphorylation. PIG3 was not detectable in p53 KO mice, which was probably due to p53 deficiency, whereas a slightly increased H2AX phosphorylation in response to patulin exposure was found in p53 KO mice (Fig. 6E). We also analyzed the histopathological changes by patulin in both p53 wild-type and knockout mice and the results are shown in Fig. S1B. No obvious pathological changes were found in the kidney tissues of both p53 wild-type (1, 3, 6 or 12 h treatment) and knockout mice (3 h treatment), suggesting continuous treatment and persistent oxidative stress may be required for a detectable pathological damage. Taken together, these data clearly supported a pro-oxidant role of p53 in response to patulin in the animal model.

Bottom Line: Patulin is a major mycotoxin found in fungal contaminated fruits and their derivative products.To the best of our knowledge, this is the first report addressing the functional role of p53 in patulin-induced oxidative stress.The findings of the present study provided novel insights into understanding mechanisms behind oxidative stress in response to patulin exposure.

View Article: PubMed Central - PubMed

Affiliation: Beijing Advanced Innovation Center for Food Nutrition and Human Health, Department of Nutrition and Health, College of Food Science and Nutritional Engineering, China Agricultural University, No17 Qinghua East Road, Haidian District, Beijing 100083, China.

ABSTRACT
Patulin is a major mycotoxin found in fungal contaminated fruits and their derivative products. Previous studies showed that patulin was able to induce increase of reactive oxygen species (ROS) generation and oxidative stress was suggested to play a pivotal role in patulin-induced multiple toxic signaling. The objective of the present study was to investigate the functional role of p53 in patulin-induced oxidative stress. Our study demonstrated that higher levels of ROS generation and DNA damage were induced in wild-type p53 cell lines than that found in either knockdown or knockout p53 cell lines in response to patulin exposure, suggesting p53 activation contributed to patulin-induced ROS generation. Mechanistically, we revealed that the pro-oxidant role of p53 in response to patulin was attributed to its ability to suppress catalase activity through up-regulation of PIG3. Moreover, these in vitro findings were further validated in the p53 wild-type/knockout mouse model. To the best of our knowledge, this is the first report addressing the functional role of p53 in patulin-induced oxidative stress. The findings of the present study provided novel insights into understanding mechanisms behind oxidative stress in response to patulin exposure.

No MeSH data available.


Related in: MedlinePlus