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Prion Protein Protects against Renal Ischemia/Reperfusion Injury.

Zhang B, Cowden D, Zhang F, Yuan J, Siedlak S, Abouelsaad M, Zeng L, Zhou X, O'Toole J, Das AS, Kofskey D, Warren M, Bian Z, Cui Y, Tan T, Kresak A, Wyza RE, Petersen RB, Wang GX, Kong Q, Wang X, Sedor J, Zhu X, Zhu H, Zou WQ - PLoS ONE (2015)

Bottom Line: The cellular prion protein (PrPC), a protein most noted for its link to prion diseases, has been found to play a protective role in ischemic brain injury.While PrP was undetectable in KO kidneys, Western blotting revealed an increase in PrP in IR-injured WT kidneys compared to sham-treated kidneys.Compared to WT, KO kidneys exhibited increases in oxidative stress markers heme oxygenase-1, nitrotyrosine, and Nε-(carboxymethyl)lysine, and decreases in mitochondrial complexes I and III.

View Article: PubMed Central - PubMed

Affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, HuBei, The People's Republic of China; Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio, United States of America; Key Laboratory of Ministry of Health and Key Laboratory of Ministry of Education, Wuhan, HuBei, The People's Republic of China.

ABSTRACT
The cellular prion protein (PrPC), a protein most noted for its link to prion diseases, has been found to play a protective role in ischemic brain injury. To investigate the role of PrPC in the kidney, an organ highly prone to ischemia/reperfusion (IR) injury, we examined wild-type (WT) and PrPC knockout (KO) mice that were subjected to 30-min of renal ischemia followed by 1, 2, or 3 days of reperfusion. Renal dysfunction and structural damage was more severe in KO than in WT mice. While PrP was undetectable in KO kidneys, Western blotting revealed an increase in PrP in IR-injured WT kidneys compared to sham-treated kidneys. Compared to WT, KO kidneys exhibited increases in oxidative stress markers heme oxygenase-1, nitrotyrosine, and Nε-(carboxymethyl)lysine, and decreases in mitochondrial complexes I and III. Notably, phosphorylated extracellular signal-regulated kinase (pERK) staining was predominantly observed in tubular cells from KO mice following 2 days of reperfusion, a time at which significant differences in renal dysfunction, histological changes, oxidative stress, and mitochondrial complexes between WT and KO mice were observed. Our study provides the first evidence that PrPC may play a protective role in renal IR injury, likely through its effects on mitochondria and ERK signaling pathways.

No MeSH data available.


Related in: MedlinePlus

Detection of HO-1, nitrotyrosine and CML of sham and IR-injured WT and KO kidneys.(A) Detection of HO-1 from the frozen renal tissues of IR-injured WT and KO mice (three each) by Western blotting. The equal amounts of total proteins from renal homogenates were loaded based on BCA protein assay and the amounts of samples loaded were monitored by determining GAPDH. The Western blots are representative of three experiments. (B) Bar graph of HO-1 intensity based on quantitative analysis of the HO-1 band by densitometric analysis. The intensity of the protein band was normalized with GAPDH band intensity in each corresponding land. *p < 0.05; **p < 0.01. (C) HO-1 immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (D) Nitrotyrosine immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (E) CML immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (original magnification, x 200).
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pone.0136923.g004: Detection of HO-1, nitrotyrosine and CML of sham and IR-injured WT and KO kidneys.(A) Detection of HO-1 from the frozen renal tissues of IR-injured WT and KO mice (three each) by Western blotting. The equal amounts of total proteins from renal homogenates were loaded based on BCA protein assay and the amounts of samples loaded were monitored by determining GAPDH. The Western blots are representative of three experiments. (B) Bar graph of HO-1 intensity based on quantitative analysis of the HO-1 band by densitometric analysis. The intensity of the protein band was normalized with GAPDH band intensity in each corresponding land. *p < 0.05; **p < 0.01. (C) HO-1 immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (D) Nitrotyrosine immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (E) CML immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (original magnification, x 200).

Mentions: Heme oxygenase-1 (HO-1) has been observed to be upregulated by stress, and is believed to mediate transient resistance to oxidative damage from IR injury [34]. Using both Western blotting and immunohistochemistry, the levels of HO-1 were determined in sham and IR-injured kidneys. Western blot analysis revealed significant increases in the HO-1 levels on day 1 in the kidneys of injured WT and KO mice compared to sham WT and KO mice (p < 0.001). HO-1 in WT kidneys decreased to the amounts similar to sham mice by days 2 and 3 (Fig 4A and 4B). Although HO-1 in the KO kidney also diminished on days 2 and 3 compared to day 1, the KO mice still exhibited significantly higher HO-1 than WT (Fig 4A and 4B). The difference in the mean value of HO-1 was highly significant among all IR-injured kidneys of both WT and KO with different reperfusion times, as analyzed by the one-way ANOVA (p < 0.001). Consistent with the Western blot, immunohistochemistry revealed extremely intense HO-1 staining in kidney tubules in KO and WT mice on day 1; however, in contrast to the Western blot, more intense HO-1 staining was generally observed in WT than in KO kidneys of sham and all IR-injured mice (Fig 4C).


Prion Protein Protects against Renal Ischemia/Reperfusion Injury.

Zhang B, Cowden D, Zhang F, Yuan J, Siedlak S, Abouelsaad M, Zeng L, Zhou X, O'Toole J, Das AS, Kofskey D, Warren M, Bian Z, Cui Y, Tan T, Kresak A, Wyza RE, Petersen RB, Wang GX, Kong Q, Wang X, Sedor J, Zhu X, Zhu H, Zou WQ - PLoS ONE (2015)

Detection of HO-1, nitrotyrosine and CML of sham and IR-injured WT and KO kidneys.(A) Detection of HO-1 from the frozen renal tissues of IR-injured WT and KO mice (three each) by Western blotting. The equal amounts of total proteins from renal homogenates were loaded based on BCA protein assay and the amounts of samples loaded were monitored by determining GAPDH. The Western blots are representative of three experiments. (B) Bar graph of HO-1 intensity based on quantitative analysis of the HO-1 band by densitometric analysis. The intensity of the protein band was normalized with GAPDH band intensity in each corresponding land. *p < 0.05; **p < 0.01. (C) HO-1 immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (D) Nitrotyrosine immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (E) CML immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (original magnification, x 200).
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getmorefigures.php?uid=PMC4556704&req=5

pone.0136923.g004: Detection of HO-1, nitrotyrosine and CML of sham and IR-injured WT and KO kidneys.(A) Detection of HO-1 from the frozen renal tissues of IR-injured WT and KO mice (three each) by Western blotting. The equal amounts of total proteins from renal homogenates were loaded based on BCA protein assay and the amounts of samples loaded were monitored by determining GAPDH. The Western blots are representative of three experiments. (B) Bar graph of HO-1 intensity based on quantitative analysis of the HO-1 band by densitometric analysis. The intensity of the protein band was normalized with GAPDH band intensity in each corresponding land. *p < 0.05; **p < 0.01. (C) HO-1 immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (D) Nitrotyrosine immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (bar = 200 μm; original magnification, x 400). (E) CML immunohistochemistry of sham and IR-injured WT and KO mice. IHC staining images are representative renal tissue sections from three mice of each group (original magnification, x 200).
Mentions: Heme oxygenase-1 (HO-1) has been observed to be upregulated by stress, and is believed to mediate transient resistance to oxidative damage from IR injury [34]. Using both Western blotting and immunohistochemistry, the levels of HO-1 were determined in sham and IR-injured kidneys. Western blot analysis revealed significant increases in the HO-1 levels on day 1 in the kidneys of injured WT and KO mice compared to sham WT and KO mice (p < 0.001). HO-1 in WT kidneys decreased to the amounts similar to sham mice by days 2 and 3 (Fig 4A and 4B). Although HO-1 in the KO kidney also diminished on days 2 and 3 compared to day 1, the KO mice still exhibited significantly higher HO-1 than WT (Fig 4A and 4B). The difference in the mean value of HO-1 was highly significant among all IR-injured kidneys of both WT and KO with different reperfusion times, as analyzed by the one-way ANOVA (p < 0.001). Consistent with the Western blot, immunohistochemistry revealed extremely intense HO-1 staining in kidney tubules in KO and WT mice on day 1; however, in contrast to the Western blot, more intense HO-1 staining was generally observed in WT than in KO kidneys of sham and all IR-injured mice (Fig 4C).

Bottom Line: The cellular prion protein (PrPC), a protein most noted for its link to prion diseases, has been found to play a protective role in ischemic brain injury.While PrP was undetectable in KO kidneys, Western blotting revealed an increase in PrP in IR-injured WT kidneys compared to sham-treated kidneys.Compared to WT, KO kidneys exhibited increases in oxidative stress markers heme oxygenase-1, nitrotyrosine, and Nε-(carboxymethyl)lysine, and decreases in mitochondrial complexes I and III.

View Article: PubMed Central - PubMed

Affiliation: Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, HuBei, The People's Republic of China; Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio, United States of America; Key Laboratory of Ministry of Health and Key Laboratory of Ministry of Education, Wuhan, HuBei, The People's Republic of China.

ABSTRACT
The cellular prion protein (PrPC), a protein most noted for its link to prion diseases, has been found to play a protective role in ischemic brain injury. To investigate the role of PrPC in the kidney, an organ highly prone to ischemia/reperfusion (IR) injury, we examined wild-type (WT) and PrPC knockout (KO) mice that were subjected to 30-min of renal ischemia followed by 1, 2, or 3 days of reperfusion. Renal dysfunction and structural damage was more severe in KO than in WT mice. While PrP was undetectable in KO kidneys, Western blotting revealed an increase in PrP in IR-injured WT kidneys compared to sham-treated kidneys. Compared to WT, KO kidneys exhibited increases in oxidative stress markers heme oxygenase-1, nitrotyrosine, and Nε-(carboxymethyl)lysine, and decreases in mitochondrial complexes I and III. Notably, phosphorylated extracellular signal-regulated kinase (pERK) staining was predominantly observed in tubular cells from KO mice following 2 days of reperfusion, a time at which significant differences in renal dysfunction, histological changes, oxidative stress, and mitochondrial complexes between WT and KO mice were observed. Our study provides the first evidence that PrPC may play a protective role in renal IR injury, likely through its effects on mitochondria and ERK signaling pathways.

No MeSH data available.


Related in: MedlinePlus