The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease.
Inflammatory cholestatic liver diseases, including Primary Sclerosing Cholangitis (PSC), are characterized by periportal inflammation with progression to cirrhosis. The objective of this study was to examine interactions between oxidative stress and autophagy in cholestasis. Using hepatic tissue fro...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0276879 |
| _version_ | 1797808292610703360 |
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| author | Colin T Shearn Aimee L Anderson Michael W Devereux David J Orlicky Cole Michel Dennis R Petersen Colin G Miller Sanjiv Harpavat Edward E Schmidt Ronald J Sokol |
| author_facet | Colin T Shearn Aimee L Anderson Michael W Devereux David J Orlicky Cole Michel Dennis R Petersen Colin G Miller Sanjiv Harpavat Edward E Schmidt Ronald J Sokol |
| author_sort | Colin T Shearn |
| collection | DOAJ |
| description | Inflammatory cholestatic liver diseases, including Primary Sclerosing Cholangitis (PSC), are characterized by periportal inflammation with progression to cirrhosis. The objective of this study was to examine interactions between oxidative stress and autophagy in cholestasis. Using hepatic tissue from male acute cholestatic (bile duct ligated) as well as chronic cholestatic (Mdr2KO) mice, localization of oxidative stress, the antioxidant response and induction of autophagy were analyzed and compared to human PSC liver. Concurrently, the ability of reactive aldehydes to post-translationally modify the autophagosome marker p62 was assessed in PSC liver tissue and in cell culture. Expression of autophagy markers was upregulated in human and mouse cholestatic liver. Whereas mRNA expression of Atg12, Lamp1, Sqstm1 and Map1lc3 was increased in acute cholestasis in mice, it was either suppressed or not significantly changed in chronic cholestasis. In human and murine cholestasis, periportal hepatocytes showed increased IHC staining of ubiquitin, 4-HNE, p62, and selected antioxidant proteins. Increased p62 staining colocalized with accumulation of 4-HNE-modified proteins in periportal parenchymal cells as well as with periportal macrophages in both human and mouse liver. Mechanistically, p62 was identified as a direct target of lipid aldehyde adduction in PSC hepatic tissue and in vitro cell culture. In vitro LS-MS/MS analysis of 4-HNE treated recombinant p62 identified carbonylation of His123, Cys128, His174, His181, Lys238, Cys290, His340, Lys341 and His385. These data indicate that dysregulation of autophagy and oxidative stress/protein damage are present in the same periportal hepatocyte compartment of both human and murine cholestasis. Thus, our results suggest that both increased expression as well as ineffective autophagic degradation of oxidatively-modified proteins contributes to injury in periportal parenchymal cells and that direct modification of p62 by reactive aldehydes may contribute to autophagic dysfunction. |
| first_indexed | 2024-03-13T06:35:26Z |
| format | Article |
| id | doaj.art-ce15a1ec4a624b35aa47d9ec04c0050d |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-03-13T06:35:26Z |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj.art-ce15a1ec4a624b35aa47d9ec04c0050d2023-06-09T05:31:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032022-01-011711e027687910.1371/journal.pone.0276879The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease.Colin T ShearnAimee L AndersonMichael W DevereuxDavid J OrlickyCole MichelDennis R PetersenColin G MillerSanjiv HarpavatEdward E SchmidtRonald J SokolInflammatory cholestatic liver diseases, including Primary Sclerosing Cholangitis (PSC), are characterized by periportal inflammation with progression to cirrhosis. The objective of this study was to examine interactions between oxidative stress and autophagy in cholestasis. Using hepatic tissue from male acute cholestatic (bile duct ligated) as well as chronic cholestatic (Mdr2KO) mice, localization of oxidative stress, the antioxidant response and induction of autophagy were analyzed and compared to human PSC liver. Concurrently, the ability of reactive aldehydes to post-translationally modify the autophagosome marker p62 was assessed in PSC liver tissue and in cell culture. Expression of autophagy markers was upregulated in human and mouse cholestatic liver. Whereas mRNA expression of Atg12, Lamp1, Sqstm1 and Map1lc3 was increased in acute cholestasis in mice, it was either suppressed or not significantly changed in chronic cholestasis. In human and murine cholestasis, periportal hepatocytes showed increased IHC staining of ubiquitin, 4-HNE, p62, and selected antioxidant proteins. Increased p62 staining colocalized with accumulation of 4-HNE-modified proteins in periportal parenchymal cells as well as with periportal macrophages in both human and mouse liver. Mechanistically, p62 was identified as a direct target of lipid aldehyde adduction in PSC hepatic tissue and in vitro cell culture. In vitro LS-MS/MS analysis of 4-HNE treated recombinant p62 identified carbonylation of His123, Cys128, His174, His181, Lys238, Cys290, His340, Lys341 and His385. These data indicate that dysregulation of autophagy and oxidative stress/protein damage are present in the same periportal hepatocyte compartment of both human and murine cholestasis. Thus, our results suggest that both increased expression as well as ineffective autophagic degradation of oxidatively-modified proteins contributes to injury in periportal parenchymal cells and that direct modification of p62 by reactive aldehydes may contribute to autophagic dysfunction.https://doi.org/10.1371/journal.pone.0276879 |
| spellingShingle | Colin T Shearn Aimee L Anderson Michael W Devereux David J Orlicky Cole Michel Dennis R Petersen Colin G Miller Sanjiv Harpavat Edward E Schmidt Ronald J Sokol The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. PLoS ONE |
| title | The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. |
| title_full | The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. |
| title_fullStr | The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. |
| title_full_unstemmed | The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. |
| title_short | The autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease. |
| title_sort | autophagic protein p62 is a target of reactive aldehydes in human and murine cholestatic liver disease |
| url | https://doi.org/10.1371/journal.pone.0276879 |
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