KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43
Abstract Background Ossification of ligamentum flavum (OLF) is a prevalent degenerative spinal disease, typically causing severe neurological dysfunction. Kruppel-like factor 5 (KLF5) plays an essential role in the regulation of skeletal development. However, the mechanism KLF5 plays in OLF remains...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-04-01
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| Series: | Journal of Orthopaedic Surgery and Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13018-024-04702-2 |
| _version_ | 1827277068982812672 |
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| author | Hubing Guo Lingxia Yang Jin Liu Liqi Chen Yufeng Huang Jinsong Li |
| author_facet | Hubing Guo Lingxia Yang Jin Liu Liqi Chen Yufeng Huang Jinsong Li |
| author_sort | Hubing Guo |
| collection | DOAJ |
| description | Abstract Background Ossification of ligamentum flavum (OLF) is a prevalent degenerative spinal disease, typically causing severe neurological dysfunction. Kruppel-like factor 5 (KLF5) plays an essential role in the regulation of skeletal development. However, the mechanism KLF5 plays in OLF remains unclear, necessitating further investigative studies. Methods qRT-PCR, immunofluorescent staining and western blot were used to measure the expression of KLF5. Alkaline Phosphatase (ALP) staining, Alizarin red staining (ARS), and the expression of Runt-related transcription factor 2 (RUNX2), osteopontin (OPN), and osteocalcin (OCN) were used to evaluate the osteogenic differentiation. Luciferase activity assay and ChIP-PCR were performed to investigate the molecular mechanisms. Results KLF5 was significantly upregulated in OLF fibroblasts in contrast to normal ligamentum flavum (LF) fibroblasts. Silencing KLF5 diminished osteogenic markers and mineralized nodules, while its overexpression had the opposite effect, confirming KLF5’s role in promoting ossification. Moreover, KLF5 promotes the ossification of LF by activating the transcription of Connexin 43 (CX43), and overexpressing CX43 could reverse the suppressive impact of KLF5 knockdown on OLF fibroblasts’ osteogenesis. Conclusion KLF5 promotes the OLF by transcriptionally activating CX43. This finding contributes significantly to our understanding of OLF and may provide new therapeutic targets. |
| first_indexed | 2024-04-24T07:14:41Z |
| format | Article |
| id | doaj.art-f9c736cafec94ae1bdd8c39e81bb5a08 |
| institution | Directory Open Access Journal |
| issn | 1749-799X |
| language | English |
| last_indexed | 2024-04-24T07:14:41Z |
| publishDate | 2024-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Orthopaedic Surgery and Research |
| spelling | doaj.art-f9c736cafec94ae1bdd8c39e81bb5a082024-04-21T11:24:59ZengBMCJournal of Orthopaedic Surgery and Research1749-799X2024-04-011911910.1186/s13018-024-04702-2KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43Hubing Guo0Lingxia Yang1Jin Liu2Liqi Chen3Yufeng Huang4Jinsong Li5The First Department of Orthopaedic Surgery, The First Hospital of TianshuiDepartment of Odermatology, The First Hospital of TianshuiThe First Department of Orthopaedic Surgery, The First Hospital of TianshuiThe First Department of Orthopaedic Surgery, The First Hospital of TianshuiThe First Department of Orthopaedic Surgery, The First Hospital of TianshuiDepartment of Spine Surgery, The Third Xiangya Hospital, Central South UniversityAbstract Background Ossification of ligamentum flavum (OLF) is a prevalent degenerative spinal disease, typically causing severe neurological dysfunction. Kruppel-like factor 5 (KLF5) plays an essential role in the regulation of skeletal development. However, the mechanism KLF5 plays in OLF remains unclear, necessitating further investigative studies. Methods qRT-PCR, immunofluorescent staining and western blot were used to measure the expression of KLF5. Alkaline Phosphatase (ALP) staining, Alizarin red staining (ARS), and the expression of Runt-related transcription factor 2 (RUNX2), osteopontin (OPN), and osteocalcin (OCN) were used to evaluate the osteogenic differentiation. Luciferase activity assay and ChIP-PCR were performed to investigate the molecular mechanisms. Results KLF5 was significantly upregulated in OLF fibroblasts in contrast to normal ligamentum flavum (LF) fibroblasts. Silencing KLF5 diminished osteogenic markers and mineralized nodules, while its overexpression had the opposite effect, confirming KLF5’s role in promoting ossification. Moreover, KLF5 promotes the ossification of LF by activating the transcription of Connexin 43 (CX43), and overexpressing CX43 could reverse the suppressive impact of KLF5 knockdown on OLF fibroblasts’ osteogenesis. Conclusion KLF5 promotes the OLF by transcriptionally activating CX43. This finding contributes significantly to our understanding of OLF and may provide new therapeutic targets.https://doi.org/10.1186/s13018-024-04702-2KLF5Ligamentum flavumOssificationCX43 |
| spellingShingle | Hubing Guo Lingxia Yang Jin Liu Liqi Chen Yufeng Huang Jinsong Li KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 Journal of Orthopaedic Surgery and Research KLF5 Ligamentum flavum Ossification CX43 |
| title | KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 |
| title_full | KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 |
| title_fullStr | KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 |
| title_full_unstemmed | KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 |
| title_short | KLF5 promotes the ossification process of ligamentum flavum by transcriptionally activating CX43 |
| title_sort | klf5 promotes the ossification process of ligamentum flavum by transcriptionally activating cx43 |
| topic | KLF5 Ligamentum flavum Ossification CX43 |
| url | https://doi.org/10.1186/s13018-024-04702-2 |
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