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PEP-1-GRX-1 Modulates Matrix Metalloproteinase-13 and Nitric Oxide Expression of Human Articular Chondrocytes.

Identifieur interne : 000333 ( Main/Exploration ); précédent : 000332; suivant : 000334

PEP-1-GRX-1 Modulates Matrix Metalloproteinase-13 and Nitric Oxide Expression of Human Articular Chondrocytes.

Auteurs : Hyun Sook Hwang ; In Young Park ; Hyun Ah Kim ; Soo Young Choi

Source :

RBID : pubmed:28214840

Descripteurs français

English descriptors

Abstract

BACKGROUND

The protein transduction domain (PTD) enables therapeutic proteins to directly penetrate the membranes of cells and tissues, and has been increasingly utilized. Glutaredoxin-1 (GRX-1) is an endogenous antioxidant enzyme involved in the cellular redox homeostasis system. In this study, we investigated whether PEP-1-GRX-1, a fusion protein of GRX-1 and PEP-1 peptide, a PTD, could suppress catabolic responses in primary human articular chondrocytes and a mouse carrageenan-induced paw edema model.

METHODS

Human articular chondrocytes were isolated enzymatically from articular cartilage and cultured in a monolayer. The transduction efficiency of PEP-1-GRX-1 into articular chondrocytes was measured by western blot and immunohistochemistry. The effects of PEP-1-GRX-1 on matrix metalloproteinases (MMPs) and catabolic factor expression in interleukin (IL)-1β- and lipopolysaccharide (LPS)-treated chondrocytes were analyzed by real-time quantitative reverse transcription-polymerase chain reaction and western blot. The effect of PEP-1-GRX1 on the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB) signaling pathway were also analyzed by western blot. Finally, the inhibitory effect of PEP-1-GRX-1 on MMP-13 production was measured in vivo in a mouse carrageenan-induced paw edema model.

RESULTS

PEP-1-GRX-1 significantly penetrated into human chondrocytes and mouse cartilage, whereas GRX-1 did not. PEP-1-GRX-1 significantly suppressed MMP-13 expression and nitric oxide (NO) production in LPS-stimulated chondrocytes, and NO production in IL-1β-stimulated chondrocytes, compared with GRX-1. In addition, PEP-1-GRX-1 decreased IL-1β- and LPS-induced activation of MAPK and NF-κB. In the mouse model of carrageenan-induced paw edema, PEP-1-GRX-1 significantly suppressed carrageenan-induced MMP-13 production as well as paw edema.

CONCLUSION

These results demonstrate that PEP-1-GRX-1 can be transduced efficiently in vitro and in vivo into human chondrocytes and mouse cartilage tissue and downregulate catabolic responses in chondrocytes by inhibiting the MAPK and NF-κB pathway. PEP-1-GRX-1 thus has the potential to reduce catabolic responses in chondrocytes and cartilage.


DOI: 10.1159/000456090
PubMed: 28214840


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Le document en format XML

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<term>Cartilage, Articular (drug effects)</term>
<term>Cartilage, Articular (metabolism)</term>
<term>Cells, Cultured (MeSH)</term>
<term>Cysteamine (analogs & derivatives)</term>
<term>Cysteamine (metabolism)</term>
<term>Disease Models, Animal (MeSH)</term>
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<term>Edema (chemically induced)</term>
<term>Edema (metabolism)</term>
<term>Edema (pathology)</term>
<term>Glutaredoxins (genetics)</term>
<term>Glutaredoxins (metabolism)</term>
<term>Humans (MeSH)</term>
<term>Immunohistochemistry (MeSH)</term>
<term>Interleukin-1beta (pharmacology)</term>
<term>Lipopolysaccharides (toxicity)</term>
<term>Male (MeSH)</term>
<term>Matrix Metalloproteinase 13 (genetics)</term>
<term>Matrix Metalloproteinase 13 (metabolism)</term>
<term>Mice (MeSH)</term>
<term>Mice, Inbred C57BL (MeSH)</term>
<term>Mitogen-Activated Protein Kinases (metabolism)</term>
<term>NF-kappa B (metabolism)</term>
<term>Nitric Oxide (metabolism)</term>
<term>Peptides (genetics)</term>
<term>Peptides (metabolism)</term>
<term>Recombinant Fusion Proteins (biosynthesis)</term>
<term>Recombinant Fusion Proteins (isolation & purification)</term>
<term>Recombinant Fusion Proteins (pharmacology)</term>
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<term>Animaux (MeSH)</term>
<term>Carragénane (toxicité)</term>
<term>Cartilage articulaire (cytologie)</term>
<term>Cartilage articulaire (effets des médicaments et des substances chimiques)</term>
<term>Cartilage articulaire (métabolisme)</term>
<term>Cellules cultivées (MeSH)</term>
<term>Facteur de transcription NF-kappa B (métabolisme)</term>
<term>Glutarédoxines (génétique)</term>
<term>Glutarédoxines (métabolisme)</term>
<term>Humains (MeSH)</term>
<term>Immunohistochimie (MeSH)</term>
<term>Interleukine-1 bêta (pharmacologie)</term>
<term>Lipopolysaccharides (toxicité)</term>
<term>Matrix Metalloproteinase 13 (génétique)</term>
<term>Matrix Metalloproteinase 13 (métabolisme)</term>
<term>Mercaptamine (analogues et dérivés)</term>
<term>Mercaptamine (métabolisme)</term>
<term>Mitogen-Activated Protein Kinases (métabolisme)</term>
<term>Modèles animaux de maladie humaine (MeSH)</term>
<term>Monoxyde d'azote (métabolisme)</term>
<term>Mâle (MeSH)</term>
<term>Oedème (anatomopathologie)</term>
<term>Oedème (induit chimiquement)</term>
<term>Oedème (métabolisme)</term>
<term>Peptides (génétique)</term>
<term>Peptides (métabolisme)</term>
<term>Protéines de fusion recombinantes (biosynthèse)</term>
<term>Protéines de fusion recombinantes (isolement et purification)</term>
<term>Protéines de fusion recombinantes (pharmacologie)</term>
<term>Régulation négative (effets des médicaments et des substances chimiques)</term>
<term>Souris (MeSH)</term>
<term>Souris de lignée C57BL (MeSH)</term>
<term>Transduction du signal (effets des médicaments et des substances chimiques)</term>
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<term>Cysteamine</term>
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<term>Recombinant Fusion Proteins</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en">
<term>Glutaredoxins</term>
<term>Matrix Metalloproteinase 13</term>
<term>Peptides</term>
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<term>Recombinant Fusion Proteins</term>
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<term>Cysteamine</term>
<term>Glutaredoxins</term>
<term>Matrix Metalloproteinase 13</term>
<term>Mitogen-Activated Protein Kinases</term>
<term>NF-kappa B</term>
<term>Nitric Oxide</term>
<term>Peptides</term>
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<term>Lipopolysaccharides</term>
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<term>Mercaptamine</term>
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<term>Oedème</term>
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<term>Protéines de fusion recombinantes</term>
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<term>Cartilage articulaire</term>
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<term>Cartilage, Articular</term>
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<keywords scheme="MESH" qualifier="drug effects" xml:lang="en">
<term>Cartilage, Articular</term>
<term>Down-Regulation</term>
<term>Signal Transduction</term>
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<keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr">
<term>Cartilage articulaire</term>
<term>Régulation négative</term>
<term>Transduction du signal</term>
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<term>Glutarédoxines</term>
<term>Matrix Metalloproteinase 13</term>
<term>Peptides</term>
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<term>Oedème</term>
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<term>Protéines de fusion recombinantes</term>
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<term>Edema</term>
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<term>Facteur de transcription NF-kappa B</term>
<term>Glutarédoxines</term>
<term>Matrix Metalloproteinase 13</term>
<term>Mercaptamine</term>
<term>Mitogen-Activated Protein Kinases</term>
<term>Monoxyde d'azote</term>
<term>Oedème</term>
<term>Peptides</term>
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<term>Cells, Cultured</term>
<term>Disease Models, Animal</term>
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<term>Cellules cultivées</term>
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<term>Immunohistochimie</term>
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<div type="abstract" xml:lang="en">
<p>
<b>BACKGROUND</b>
</p>
<p>The protein transduction domain (PTD) enables therapeutic proteins to directly penetrate the membranes of cells and tissues, and has been increasingly utilized. Glutaredoxin-1 (GRX-1) is an endogenous antioxidant enzyme involved in the cellular redox homeostasis system. In this study, we investigated whether PEP-1-GRX-1, a fusion protein of GRX-1 and PEP-1 peptide, a PTD, could suppress catabolic responses in primary human articular chondrocytes and a mouse carrageenan-induced paw edema model.</p>
</div>
<div type="abstract" xml:lang="en">
<p>
<b>METHODS</b>
</p>
<p>Human articular chondrocytes were isolated enzymatically from articular cartilage and cultured in a monolayer. The transduction efficiency of PEP-1-GRX-1 into articular chondrocytes was measured by western blot and immunohistochemistry. The effects of PEP-1-GRX-1 on matrix metalloproteinases (MMPs) and catabolic factor expression in interleukin (IL)-1β- and lipopolysaccharide (LPS)-treated chondrocytes were analyzed by real-time quantitative reverse transcription-polymerase chain reaction and western blot. The effect of PEP-1-GRX1 on the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB) signaling pathway were also analyzed by western blot. Finally, the inhibitory effect of PEP-1-GRX-1 on MMP-13 production was measured in vivo in a mouse carrageenan-induced paw edema model.</p>
</div>
<div type="abstract" xml:lang="en">
<p>
<b>RESULTS</b>
</p>
<p>PEP-1-GRX-1 significantly penetrated into human chondrocytes and mouse cartilage, whereas GRX-1 did not. PEP-1-GRX-1 significantly suppressed MMP-13 expression and nitric oxide (NO) production in LPS-stimulated chondrocytes, and NO production in IL-1β-stimulated chondrocytes, compared with GRX-1. In addition, PEP-1-GRX-1 decreased IL-1β- and LPS-induced activation of MAPK and NF-κB. In the mouse model of carrageenan-induced paw edema, PEP-1-GRX-1 significantly suppressed carrageenan-induced MMP-13 production as well as paw edema.</p>
</div>
<div type="abstract" xml:lang="en">
<p>
<b>CONCLUSION</b>
</p>
<p>These results demonstrate that PEP-1-GRX-1 can be transduced efficiently in vitro and in vivo into human chondrocytes and mouse cartilage tissue and downregulate catabolic responses in chondrocytes by inhibiting the MAPK and NF-κB pathway. PEP-1-GRX-1 thus has the potential to reduce catabolic responses in chondrocytes and cartilage.</p>
</div>
</front>
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<AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">The protein transduction domain (PTD) enables therapeutic proteins to directly penetrate the membranes of cells and tissues, and has been increasingly utilized. Glutaredoxin-1 (GRX-1) is an endogenous antioxidant enzyme involved in the cellular redox homeostasis system. In this study, we investigated whether PEP-1-GRX-1, a fusion protein of GRX-1 and PEP-1 peptide, a PTD, could suppress catabolic responses in primary human articular chondrocytes and a mouse carrageenan-induced paw edema model.</AbstractText>
<AbstractText Label="METHODS" NlmCategory="METHODS">Human articular chondrocytes were isolated enzymatically from articular cartilage and cultured in a monolayer. The transduction efficiency of PEP-1-GRX-1 into articular chondrocytes was measured by western blot and immunohistochemistry. The effects of PEP-1-GRX-1 on matrix metalloproteinases (MMPs) and catabolic factor expression in interleukin (IL)-1β- and lipopolysaccharide (LPS)-treated chondrocytes were analyzed by real-time quantitative reverse transcription-polymerase chain reaction and western blot. The effect of PEP-1-GRX1 on the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB) signaling pathway were also analyzed by western blot. Finally, the inhibitory effect of PEP-1-GRX-1 on MMP-13 production was measured in vivo in a mouse carrageenan-induced paw edema model.</AbstractText>
<AbstractText Label="RESULTS" NlmCategory="RESULTS">PEP-1-GRX-1 significantly penetrated into human chondrocytes and mouse cartilage, whereas GRX-1 did not. PEP-1-GRX-1 significantly suppressed MMP-13 expression and nitric oxide (NO) production in LPS-stimulated chondrocytes, and NO production in IL-1β-stimulated chondrocytes, compared with GRX-1. In addition, PEP-1-GRX-1 decreased IL-1β- and LPS-induced activation of MAPK and NF-κB. In the mouse model of carrageenan-induced paw edema, PEP-1-GRX-1 significantly suppressed carrageenan-induced MMP-13 production as well as paw edema.</AbstractText>
<AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">These results demonstrate that PEP-1-GRX-1 can be transduced efficiently in vitro and in vivo into human chondrocytes and mouse cartilage tissue and downregulate catabolic responses in chondrocytes by inhibiting the MAPK and NF-κB pathway. PEP-1-GRX-1 thus has the potential to reduce catabolic responses in chondrocytes and cartilage.</AbstractText>
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<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName>
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<MeshHeading>
<DescriptorName UI="D016328" MajorTopicYN="N">NF-kappa B</DescriptorName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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<MeshHeading>
<DescriptorName UI="D009569" MajorTopicYN="N">Nitric Oxide</DescriptorName>
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<MeshHeading>
<DescriptorName UI="D010455" MajorTopicYN="N">Peptides</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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<DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName>
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<QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName>
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<DescriptorName UI="D015398" MajorTopicYN="N">Signal Transduction</DescriptorName>
<QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName>
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<KeywordList Owner="NOTNLM">
<Keyword MajorTopicYN="N">Chondrocyte</Keyword>
<Keyword MajorTopicYN="N">Glutaredoxin-1</Keyword>
<Keyword MajorTopicYN="N">Matrix metalloproteinase</Keyword>
<Keyword MajorTopicYN="N">Osteoarthritis.</Keyword>
<Keyword MajorTopicYN="N">Protein transduction domain</Keyword>
</KeywordList>
</MedlineCitation>
<PubmedData>
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<PubMedPubDate PubStatus="received">
<Year>2016</Year>
<Month>08</Month>
<Day>08</Day>
</PubMedPubDate>
<PubMedPubDate PubStatus="accepted">
<Year>2016</Year>
<Month>11</Month>
<Day>29</Day>
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<Month>2</Month>
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<Year>2017</Year>
<Month>6</Month>
<Day>7</Day>
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<Month>2</Month>
<Day>20</Day>
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<ArticleId IdType="pubmed">28214840</ArticleId>
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<name sortKey="Choi, Soo Young" sort="Choi, Soo Young" uniqKey="Choi S" first="Soo Young" last="Choi">Soo Young Choi</name>
<name sortKey="Hwang, Hyun Sook" sort="Hwang, Hyun Sook" uniqKey="Hwang H" first="Hyun Sook" last="Hwang">Hyun Sook Hwang</name>
<name sortKey="Kim, Hyun Ah" sort="Kim, Hyun Ah" uniqKey="Kim H" first="Hyun Ah" last="Kim">Hyun Ah Kim</name>
<name sortKey="Park, In Young" sort="Park, In Young" uniqKey="Park I" first="In Young" last="Park">In Young Park</name>
</noCountry>
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