Induction of interleukin-1 in articular cartilage by explantation and cutting.
OBJECTIVE: To investigate the effect of explantation and fine cutting of articular cartilage upon intracellular inflammatory signaling pathways and expression of interleukin-1 (IL-1). METHODS: Cartilage from porcine metacarpophalangeal joints was cultured in serum-free medium. Tissue extracts were...
| मुख्य लेखकों: | , , , , , |
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| स्वरूप: | Journal article |
| भाषा: | English |
| प्रकाशित: |
2004
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| _version_ | 1826270887912931328 |
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| author | Gruber, J Vincent, T Hermansson, M Bolton, M Wait, R Saklatvala, J |
| author_facet | Gruber, J Vincent, T Hermansson, M Bolton, M Wait, R Saklatvala, J |
| author_sort | Gruber, J |
| collection | OXFORD |
| description | OBJECTIVE: To investigate the effect of explantation and fine cutting of articular cartilage upon intracellular inflammatory signaling pathways and expression of interleukin-1 (IL-1). METHODS: Cartilage from porcine metacarpophalangeal joints was cultured in serum-free medium. Tissue extracts were examined for ERK activation by phosphorylated-Western blotting, for JNK and p38 MAPK activity by kinase assay, and for IkappaBalpha. IL-1alpha and IL-1beta messenger RNA (mRNA) was measured by reverse transcriptase-polymerase chain reaction. IL-1 activity was measured by the induction of serum amyloid A protein in cultured chondrocytes. RESULTS: All 3 MAPKs (p38, JNK, and ERK) were rapidly activated upon dissection and explantation of the cartilage. IL-1alpha and IL-1beta mRNA was also induced: the speed and magnitude of induction were increased if the explants had been finely cut. IL-1 activity that could be inhibited by IL-1 receptor antagonist or antibodies to IL-1alpha was found in extracts of explants cultured for 20 hours or lysates of cells isolated from them. This activity was likely due to intracellular proIL-1alpha that was not secreted. ProIL-1beta would not be detected because it is biologically inactive. The mechanism of inflammatory signaling pathway activation underlying the induction of IL-1 is unknown. CONCLUSION: Explantation and cutting of articular cartilage activates intracellular inflammatory signaling pathways and induces expression of mRNA for IL-1alpha and IL-1beta. Biologically active IL-1alpha protein was detectable in cartilage lysates and was probably intracellular proIL-1alpha. We were unable to show that IL-1 was secreted by chondrocytes. |
| first_indexed | 2024-03-06T21:47:55Z |
| format | Journal article |
| id | oxford-uuid:4a3f6a6a-4e89-4a05-bdc0-3a96de65a9b0 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T21:47:55Z |
| publishDate | 2004 |
| record_format | dspace |
| spelling | oxford-uuid:4a3f6a6a-4e89-4a05-bdc0-3a96de65a9b02022-03-26T15:36:22ZInduction of interleukin-1 in articular cartilage by explantation and cutting.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:4a3f6a6a-4e89-4a05-bdc0-3a96de65a9b0EnglishSymplectic Elements at Oxford2004Gruber, JVincent, THermansson, MBolton, MWait, RSaklatvala, J OBJECTIVE: To investigate the effect of explantation and fine cutting of articular cartilage upon intracellular inflammatory signaling pathways and expression of interleukin-1 (IL-1). METHODS: Cartilage from porcine metacarpophalangeal joints was cultured in serum-free medium. Tissue extracts were examined for ERK activation by phosphorylated-Western blotting, for JNK and p38 MAPK activity by kinase assay, and for IkappaBalpha. IL-1alpha and IL-1beta messenger RNA (mRNA) was measured by reverse transcriptase-polymerase chain reaction. IL-1 activity was measured by the induction of serum amyloid A protein in cultured chondrocytes. RESULTS: All 3 MAPKs (p38, JNK, and ERK) were rapidly activated upon dissection and explantation of the cartilage. IL-1alpha and IL-1beta mRNA was also induced: the speed and magnitude of induction were increased if the explants had been finely cut. IL-1 activity that could be inhibited by IL-1 receptor antagonist or antibodies to IL-1alpha was found in extracts of explants cultured for 20 hours or lysates of cells isolated from them. This activity was likely due to intracellular proIL-1alpha that was not secreted. ProIL-1beta would not be detected because it is biologically inactive. The mechanism of inflammatory signaling pathway activation underlying the induction of IL-1 is unknown. CONCLUSION: Explantation and cutting of articular cartilage activates intracellular inflammatory signaling pathways and induces expression of mRNA for IL-1alpha and IL-1beta. Biologically active IL-1alpha protein was detectable in cartilage lysates and was probably intracellular proIL-1alpha. We were unable to show that IL-1 was secreted by chondrocytes. |
| spellingShingle | Gruber, J Vincent, T Hermansson, M Bolton, M Wait, R Saklatvala, J Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title | Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title_full | Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title_fullStr | Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title_full_unstemmed | Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title_short | Induction of interleukin-1 in articular cartilage by explantation and cutting. |
| title_sort | induction of interleukin 1 in articular cartilage by explantation and cutting |
| work_keys_str_mv | AT gruberj inductionofinterleukin1inarticularcartilagebyexplantationandcutting AT vincentt inductionofinterleukin1inarticularcartilagebyexplantationandcutting AT hermanssonm inductionofinterleukin1inarticularcartilagebyexplantationandcutting AT boltonm inductionofinterleukin1inarticularcartilagebyexplantationandcutting AT waitr inductionofinterleukin1inarticularcartilagebyexplantationandcutting AT saklatvalaj inductionofinterleukin1inarticularcartilagebyexplantationandcutting |