title: TNF-alpha-induced NF-kappaB signaling reverses age-related declines in VEGF induction and angiogenic activity in intervertebral disc tissues.
abstract: We previously demonstrated that VEGF and its receptors were expressed in human herniated discs (HD). TNF-alpha induced VEGF, resulting in neovascularization of disc tissues in a model of HD. The goal of the current research was to investigate the precise role of TNF-alpha-induced VEGF and the mechanism of angiogenesis in disc tissues. We performed ELISAs, Western blots, and immunohistological examinations to assess the role of TNF-alpha-induced VEGF using organ disc cultures with wild type, TNF receptor 1-null (TNF-RI(null)), or TNF receptor 2-null (TNF-RII(null)) mice. VEGF induction was inhibited when we used TNF-RI(null)-derived disc tissues. NF-kappaB pathway inhibitors also strongly suppressed VEGF induction. Thus, TNF-alpha induced VEGF expression in disc cells primarily through the NF-kappaB pathway. In addition, VEGF immunoreactivity was detected predominantly in annulus fibrosus cells and increased after TNF-alpha stimulation. TNF-alpha treatment also resulted in CD31 expression on endothelial cells and formation of an anastomosing network. In contrast, angiogenic activity was strongly inhibited in the presence of NF-kappaB inhibitors or anti-VEGF antibody. Our data show angiogenesis activity in disc tissues is regulated by VEGF and the NF-kappaB pathway, both of which are induced by TNF-alpha. The level of angiogenic activity in disc tissues was closely related to aging. Because neovascularization of HD is indispensable for HD resorption, the prognosis of HD and the rate of the resorption process in patients may vary as a function of the patient's age.
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Yamanashi, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ohba
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Haro
authors.forename: Takashi
authors.initials: T
authors.lastname: Ando
authors.forename: Masanori
authors.initials: M
authors.lastname: Wako
authors.forename: Fumiko
authors.initials: F
authors.lastname: Suenaga
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Aso
authors.forename: Kensuke
authors.initials: K
authors.lastname: Koyama
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Hamada
authors.forename: Atsuhito
authors.initials: A
authors.lastname: Nakao
mesh_terms: D000375:Aging / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D007403:Intervertebral Disc / Q000098:blood supply / Q000166:cytology / Q000378:metabolism*; D007405:Intervertebral Disc Displacement / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D016328:NF-kappa B / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000502:physiology; D009924:Organ Culture Techniques; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism* / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D016328:NF-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; C506236:Tnfrsf1a protein, mouse; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse