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Experimental nonalcoholic fatty liver disease in mice leads to cytochrome p450 2a5 upregulation through nuclear factor erythroid 2-like 2 translocation.

Cui Y, Wang Q, Li X, Zhang X - Redox Biol (2013)

Bottom Line: Livers of Nrf2 (-/-) mice on the high-fat diet exhibited more oxidative stress than their wild-type counterparts as assessed by a significant depletion of reduced glutathione that was coupled with increases in malondialdehyde.Furthermore, results in Nrf2-deficient mice showed that CYP2A5 expression was significantly attenuated in the absence of Nrf2, as was found with the conventional target genes of Nrf2.These findings suggest that the high-fat diet induced alteration in cellular redox status and induction of CYP2A5 was modulated through the redox-sensitive transcription Nrf2.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin 150030, Heilongjiang, China ; College of Animal Sciences and Technology, Heilongjiang Bayi Agricultural University, 2# Xinyang Road, New Development District, Daqing 163319, Heilongjiang, China.

ABSTRACT
Mouse cytochrome P450 2A5 (CYP2A5) is upregulated in various liver diseases and a putative common feature for all of these conditions is altered cellular redox status. Nuclear factor erythroid 2-like 2 (Nrf2) is a transcription factor that is post-translationally regulated by oxidative stress and controls the transcription of protective target genes. In the present study, we have characterized the regulation of CYP2A5 by Nrf2 and evaluated gene expression, protein content and activity of anti-oxidant enzymes in the Nrf2 (+/+) and Nrf2 (-/-) mice model of non-alcoholic fatty liver (NAFLD). After eight weeks of feeding on a high-fat diet, livers from Nrf2 (-/-) mice showed a substantial increase in macro and microvesicular steatosis and a massive increase in the number of neutrophil polymorphs, compared to livers from wild-type mice treated similarly. Livers of Nrf2 (-/-) mice on the high-fat diet exhibited more oxidative stress than their wild-type counterparts as assessed by a significant depletion of reduced glutathione that was coupled with increases in malondialdehyde. Furthermore, results in Nrf2-deficient mice showed that CYP2A5 expression was significantly attenuated in the absence of Nrf2, as was found with the conventional target genes of Nrf2. The treatment of wild-type mice with high-fat diet leaded to nuclear accumulation of Nrf2, and co-immunoprecipitation experiments showed that Nrf2 was bound to Cyp2a5. These findings suggest that the high-fat diet induced alteration in cellular redox status and induction of CYP2A5 was modulated through the redox-sensitive transcription Nrf2.

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Nrf2 protein expresses along with Cyp2a5 protein interactively in WT mice and NAFLD induces the two protein expression. (A) Co-immunoprecipitated with anti-Nrf2 and Western blot with anti-Cpy2a5 showing expression of Cyp2a5. (B) Co-immunoprecipitated with anti-Cpy2a5 and Western blot with anti-Nrf2 showing expression of Nrf2. Relative amount of the co-immunoprecipitation complex Nrf2 and Cyp2a5 valuated by the ratio of band optical density. All values represent the mean±SD (n=3) against the control levels. Mean differences are significant from control group at P<0.05(⁎) and P<0.01(⁎⁎). (1) liver extracts (input) were used as positive control; (2) immunoprecipitates from wild-type control group mice livers; (3) immunoprecipitates from high-fat diet group mice livers; (4) immunoprecipitates with IgG used as negative control; IP, co-immunoprecipitation; IB: immunoblot.
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f0030: Nrf2 protein expresses along with Cyp2a5 protein interactively in WT mice and NAFLD induces the two protein expression. (A) Co-immunoprecipitated with anti-Nrf2 and Western blot with anti-Cpy2a5 showing expression of Cyp2a5. (B) Co-immunoprecipitated with anti-Cpy2a5 and Western blot with anti-Nrf2 showing expression of Nrf2. Relative amount of the co-immunoprecipitation complex Nrf2 and Cyp2a5 valuated by the ratio of band optical density. All values represent the mean±SD (n=3) against the control levels. Mean differences are significant from control group at P<0.05(⁎) and P<0.01(⁎⁎). (1) liver extracts (input) were used as positive control; (2) immunoprecipitates from wild-type control group mice livers; (3) immunoprecipitates from high-fat diet group mice livers; (4) immunoprecipitates with IgG used as negative control; IP, co-immunoprecipitation; IB: immunoblot.

Mentions: As shown in Fig. 6, Cyp2a5 protein was detected in the protein complex from the imunoprecipitation of cell lysates using an Nrf2 antibody (Fig. 6A), and Nrf2 protein also was detected in the imunoprecipitations of Cyp2a5 (Fig. 6B). Furthermore, in the high-fat diet group both Nrf2 and Cyp2a5 protein expression were higher significantly (P<0.05 or P<0.01) than in control group mice. There was no co-immunoprecipitation of Nrf2 with Cyp2a5 in the absence of antibody.


Experimental nonalcoholic fatty liver disease in mice leads to cytochrome p450 2a5 upregulation through nuclear factor erythroid 2-like 2 translocation.

Cui Y, Wang Q, Li X, Zhang X - Redox Biol (2013)

Nrf2 protein expresses along with Cyp2a5 protein interactively in WT mice and NAFLD induces the two protein expression. (A) Co-immunoprecipitated with anti-Nrf2 and Western blot with anti-Cpy2a5 showing expression of Cyp2a5. (B) Co-immunoprecipitated with anti-Cpy2a5 and Western blot with anti-Nrf2 showing expression of Nrf2. Relative amount of the co-immunoprecipitation complex Nrf2 and Cyp2a5 valuated by the ratio of band optical density. All values represent the mean±SD (n=3) against the control levels. Mean differences are significant from control group at P<0.05(⁎) and P<0.01(⁎⁎). (1) liver extracts (input) were used as positive control; (2) immunoprecipitates from wild-type control group mice livers; (3) immunoprecipitates from high-fat diet group mice livers; (4) immunoprecipitates with IgG used as negative control; IP, co-immunoprecipitation; IB: immunoblot.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814957&req=5

f0030: Nrf2 protein expresses along with Cyp2a5 protein interactively in WT mice and NAFLD induces the two protein expression. (A) Co-immunoprecipitated with anti-Nrf2 and Western blot with anti-Cpy2a5 showing expression of Cyp2a5. (B) Co-immunoprecipitated with anti-Cpy2a5 and Western blot with anti-Nrf2 showing expression of Nrf2. Relative amount of the co-immunoprecipitation complex Nrf2 and Cyp2a5 valuated by the ratio of band optical density. All values represent the mean±SD (n=3) against the control levels. Mean differences are significant from control group at P<0.05(⁎) and P<0.01(⁎⁎). (1) liver extracts (input) were used as positive control; (2) immunoprecipitates from wild-type control group mice livers; (3) immunoprecipitates from high-fat diet group mice livers; (4) immunoprecipitates with IgG used as negative control; IP, co-immunoprecipitation; IB: immunoblot.
Mentions: As shown in Fig. 6, Cyp2a5 protein was detected in the protein complex from the imunoprecipitation of cell lysates using an Nrf2 antibody (Fig. 6A), and Nrf2 protein also was detected in the imunoprecipitations of Cyp2a5 (Fig. 6B). Furthermore, in the high-fat diet group both Nrf2 and Cyp2a5 protein expression were higher significantly (P<0.05 or P<0.01) than in control group mice. There was no co-immunoprecipitation of Nrf2 with Cyp2a5 in the absence of antibody.

Bottom Line: Livers of Nrf2 (-/-) mice on the high-fat diet exhibited more oxidative stress than their wild-type counterparts as assessed by a significant depletion of reduced glutathione that was coupled with increases in malondialdehyde.Furthermore, results in Nrf2-deficient mice showed that CYP2A5 expression was significantly attenuated in the absence of Nrf2, as was found with the conventional target genes of Nrf2.These findings suggest that the high-fat diet induced alteration in cellular redox status and induction of CYP2A5 was modulated through the redox-sensitive transcription Nrf2.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin 150030, Heilongjiang, China ; College of Animal Sciences and Technology, Heilongjiang Bayi Agricultural University, 2# Xinyang Road, New Development District, Daqing 163319, Heilongjiang, China.

ABSTRACT
Mouse cytochrome P450 2A5 (CYP2A5) is upregulated in various liver diseases and a putative common feature for all of these conditions is altered cellular redox status. Nuclear factor erythroid 2-like 2 (Nrf2) is a transcription factor that is post-translationally regulated by oxidative stress and controls the transcription of protective target genes. In the present study, we have characterized the regulation of CYP2A5 by Nrf2 and evaluated gene expression, protein content and activity of anti-oxidant enzymes in the Nrf2 (+/+) and Nrf2 (-/-) mice model of non-alcoholic fatty liver (NAFLD). After eight weeks of feeding on a high-fat diet, livers from Nrf2 (-/-) mice showed a substantial increase in macro and microvesicular steatosis and a massive increase in the number of neutrophil polymorphs, compared to livers from wild-type mice treated similarly. Livers of Nrf2 (-/-) mice on the high-fat diet exhibited more oxidative stress than their wild-type counterparts as assessed by a significant depletion of reduced glutathione that was coupled with increases in malondialdehyde. Furthermore, results in Nrf2-deficient mice showed that CYP2A5 expression was significantly attenuated in the absence of Nrf2, as was found with the conventional target genes of Nrf2. The treatment of wild-type mice with high-fat diet leaded to nuclear accumulation of Nrf2, and co-immunoprecipitation experiments showed that Nrf2 was bound to Cyp2a5. These findings suggest that the high-fat diet induced alteration in cellular redox status and induction of CYP2A5 was modulated through the redox-sensitive transcription Nrf2.

Show MeSH
Related in: MedlinePlus