ABSTRACT
Low back pain (LBP) is one of the most common complains in orthopedic outpatient department and intervertebral disc degeneration (IDD) is one of the most important reasons of LBP. The mechanisms of IDD contain a complex biochemical cascade which includes inflammation, vascular ingrowth, and results in degradation of matrix. In our study, we used both in vitro and in vivo models to investigate the relation between tissue inhibitor of metalloproteinase-3 (TIMP3) expression and IDD. Loss of TIMP3 expression was found in degenerative intervertebral disc (IVD), this change of expression was closely related with the dephosphorylation of smad2/3. Overexpression of TIMP3 significantly inhibited the release of TNF-α and matrix degradation induced by Lipopolysaccharide. Vascular ingrowth was also suppressed by TIMP3 in the in vitro and in vivo models. Further, animal experiments confirmed that the degeneration of IVD was reduced after overexpression of TIMP3 in nucleus pulposus. Taken together, our results indicated TIMP-3 might play an important role in the pathogenesis of IDD and therefore be a potential therapeutic target in the future.
Subject(s)
Extracellular Matrix/pathology , Inflammation/pathology , Intervertebral Disc Degeneration/pathology , Neovascularization, Pathologic/pathology , Nucleus Pulposus/pathology , Tissue Inhibitor of Metalloproteinase-3/deficiency , Adult , Aged , Animals , Cells, Cultured , Extracellular Matrix/metabolism , Female , Humans , Inflammation/genetics , Inflammation/metabolism , Intervertebral Disc Degeneration/genetics , Intervertebral Disc Degeneration/metabolism , Male , Middle Aged , Neovascularization, Pathologic/genetics , Neovascularization, Pathologic/metabolism , Nucleus Pulposus/blood supply , Nucleus Pulposus/metabolism , Prognosis , Rats, Sprague-Dawley , Signal Transduction , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Approximately 50% of the cases of low back pain (LBP) are attributed to discogenic origin. The causes of discogenic pain are complicated and consist of a complex biochemical cascade. Neovascularization of intervertebral discs (IVDs) is believed to be associated with discogenic pain. The antiangiogenesis ability of tissue inhibitor of metalloproteinase3 (TIMP3) has been reported in many tumors, yet whether TIMP3 is associated with neovascularization of IVDs remains unknown. In the present study, both in vitro and in vivo models were used to investigate the association between discogenic pain and TIMP3 expression in nucleus pulposus (NP). PCR results demonstrated that inflammation induced downregulation of TIMP3 expression in NP cells. By using an adenovirus system to upregulate TIMP3 expression, the effect of TIMP3 on angiogenesis was measured by endothelial cell migration and tube formation assays. The results demonstrated that overexpression of TIMP3 suppressed angiogenesis in NP without the regulation of vascular endothelial growth factor (VEGF) expression. TNFα converting enzyme (TACE) expression was downregulated by TIMP3, thus inhibiting the TACEinduced activation of TNFα in NP cells. Immunohistochemical staining of IVDs also confirmed that TIMP3 inhibited the expression of substance P in NP. Taken together, the present results indicated the expression of TIMP3 in NP may have a key role in the development of discogenic pain.