TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model
Abstract Background Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Following disc injury, nerve growth factor (NGF) concentrations rise in IVDs, and anti-NGF therapy has been shown to attenuate LBP in humans. Increased levels of tumor necrosis factor-α (TNF-α) and tr...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-07-01
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| Series: | BMC Musculoskeletal Disorders |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12891-021-04509-w |
| _version_ | 1831747452052439040 |
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| author | Yuji Yokozeki Kentaro Uchida Ayumu Kawakubo Mitsufumi Nakawaki Tadashi Okubo Masayuki Miyagi Gen Inoue Makoto Itakura Hiroyuki Sekiguchi Masashi Takaso |
| author_facet | Yuji Yokozeki Kentaro Uchida Ayumu Kawakubo Mitsufumi Nakawaki Tadashi Okubo Masayuki Miyagi Gen Inoue Makoto Itakura Hiroyuki Sekiguchi Masashi Takaso |
| author_sort | Yuji Yokozeki |
| collection | DOAJ |
| description | Abstract Background Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Following disc injury, nerve growth factor (NGF) concentrations rise in IVDs, and anti-NGF therapy has been shown to attenuate LBP in humans. Increased levels of tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) in degenerative IVDs and in in vitro studies suggest that these factors promote NGF production. However, whether these factors regulate NGF in vivo remains unclear. Thus, we studied NGF regulation in a mouse model of IVD injury. Methods After inducing IVD injury, we examined mRNA levels of Tnfa, Tgfb, and Ngf in IVDs from control and IVD-injured mice across 7 days. To do this, we used magnetic cell separation to isolate CD11b ( +) (macrophage-rich) and CD11b (-) (IVD cell-rich) cell fractions from injured IVDs. To study the effect of TNF-α on Ngf expression, we examined Ngf expression in injured IVDs from C57BL/6 J and Tnfa-knockout (KO) mice (C57BL/6 J background). To study the effect of TGF-β on Ngf expression, C57/BL6J mice were given an intraperitoneal injection of either the TGF-β inhibitor SB431542 or DMSO solution (vehicle) one and two days before harvesting IVDs. Results mRNA expression of Tnfa, Tgfb, and Ngf was significantly increased in injured IVDs. Tnfa was predominantly expressed in the CD11b ( +) fraction, and Tgfb in the CD11b (-) fraction. Ngf expression was comparable between CD11b ( +) and CD11b (-) fractions, and between wild-type and Tnfa-KO mice at post-injury day (PID) 1, 3, and 7. SB431542 suppressed TGF-β-mediated Ngf expression and NGF production in vitro. Further, administration of SB431542 significantly reduced Ngf expression in IVDs such that levels were below those observed in vehicle-treated animals at PID3 and PID7. Conclusion A TGF-β inhibitor reduced Ngf expression in a mouse model of IVD injury, suggesting that TGF-β may regulate NGF expression in vivo. |
| first_indexed | 2024-12-21T21:32:57Z |
| format | Article |
| id | doaj.art-4883f062c4c54a20b245cdeaa73fb551 |
| institution | Directory Open Access Journal |
| issn | 1471-2474 |
| language | English |
| last_indexed | 2024-12-21T21:32:57Z |
| publishDate | 2021-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Musculoskeletal Disorders |
| spelling | doaj.art-4883f062c4c54a20b245cdeaa73fb5512022-12-21T18:49:34ZengBMCBMC Musculoskeletal Disorders1471-24742021-07-0122111010.1186/s12891-021-04509-wTGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury modelYuji Yokozeki0Kentaro Uchida1Ayumu Kawakubo2Mitsufumi Nakawaki3Tadashi Okubo4Masayuki Miyagi5Gen Inoue6Makoto Itakura7Hiroyuki Sekiguchi8Masashi Takaso9Department of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Laboratory Animal Science, Kitasato University School of MedicineDepartment of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Orthopaedic Surgery, Kitasato University School of MedicineDepartment of Biochemistry, Kitasato University School of MedicineShonan University of Medical Sciences Research InstituteDepartment of Orthopaedic Surgery, Kitasato University School of MedicineAbstract Background Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Following disc injury, nerve growth factor (NGF) concentrations rise in IVDs, and anti-NGF therapy has been shown to attenuate LBP in humans. Increased levels of tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) in degenerative IVDs and in in vitro studies suggest that these factors promote NGF production. However, whether these factors regulate NGF in vivo remains unclear. Thus, we studied NGF regulation in a mouse model of IVD injury. Methods After inducing IVD injury, we examined mRNA levels of Tnfa, Tgfb, and Ngf in IVDs from control and IVD-injured mice across 7 days. To do this, we used magnetic cell separation to isolate CD11b ( +) (macrophage-rich) and CD11b (-) (IVD cell-rich) cell fractions from injured IVDs. To study the effect of TNF-α on Ngf expression, we examined Ngf expression in injured IVDs from C57BL/6 J and Tnfa-knockout (KO) mice (C57BL/6 J background). To study the effect of TGF-β on Ngf expression, C57/BL6J mice were given an intraperitoneal injection of either the TGF-β inhibitor SB431542 or DMSO solution (vehicle) one and two days before harvesting IVDs. Results mRNA expression of Tnfa, Tgfb, and Ngf was significantly increased in injured IVDs. Tnfa was predominantly expressed in the CD11b ( +) fraction, and Tgfb in the CD11b (-) fraction. Ngf expression was comparable between CD11b ( +) and CD11b (-) fractions, and between wild-type and Tnfa-KO mice at post-injury day (PID) 1, 3, and 7. SB431542 suppressed TGF-β-mediated Ngf expression and NGF production in vitro. Further, administration of SB431542 significantly reduced Ngf expression in IVDs such that levels were below those observed in vehicle-treated animals at PID3 and PID7. Conclusion A TGF-β inhibitor reduced Ngf expression in a mouse model of IVD injury, suggesting that TGF-β may regulate NGF expression in vivo.https://doi.org/10.1186/s12891-021-04509-wTransforming growth factor-βNerve growth factorIntervertebral disc |
| spellingShingle | Yuji Yokozeki Kentaro Uchida Ayumu Kawakubo Mitsufumi Nakawaki Tadashi Okubo Masayuki Miyagi Gen Inoue Makoto Itakura Hiroyuki Sekiguchi Masashi Takaso TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model BMC Musculoskeletal Disorders Transforming growth factor-β Nerve growth factor Intervertebral disc |
| title | TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| title_full | TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| title_fullStr | TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| title_full_unstemmed | TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| title_short | TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| title_sort | tgf β regulates nerve growth factor expression in a mouse intervertebral disc injury model |
| topic | Transforming growth factor-β Nerve growth factor Intervertebral disc |
| url | https://doi.org/10.1186/s12891-021-04509-w |
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