High-dose TGF-β1 degrades human nucleus pulposus cells via ALK1-Smad 1/5/8 activation

Transforming growth factor beta 1 (TGF-beta 1) can promote the proliferation and differentiation of intervertebral disc cells and participates in its repair process. However, whether TGF-beta 1 engages in the process of disc degeneration has not yet been fully elucidated. The present study aimed to investigate the function of high-dose TGF-beta 1 on the metabolism of nucleus pulposus cells (NPCs). TGF-beta 1 levels in human degenerative intervertebral disc tissues and tumor necrosis factor (TNF)-alpha-induced degenerative NPCs were analyzed. Furthermore, NPCs were treated with TGF-beta 1 and inhibitors of TGF-beta 1 receptors [ALK tyrosine kinase receptor (ALK) 1 and ALK5] to determine the effect of the receptors in the mediation of NPC degeneration. The NPC state was determined by the components of secretory collagen I/II, tissue inhibitor of metalloproteinase-3 (TIMP-3) and matrix metalloproteinase (MMP)-13. The mRNA expression of Smad1/2/3/5/8, the downstream gene of TGF-beta 1 mediated by ALK, was also measured. Results showed that TGF-beta 1 and ALK1 were positively associated with the degree of degeneration of NP or NPCs in vitro, but negatively associated with ALK5. Furthermore, high-doses of TGF-(31 suppressed collagen II, but enhanced collagen I, TIMP-3, MMP-13, ALK1/5 and Smad1/2/3/5/8 expression. ALK5 inhibition induced the suppression of Smad2/3 and aggravated high-dose TGF-beta 1-induced NPC degeneration, as shown by the reduction in collagen II and increase in collagen I, TIMP-3 and MMP-13. By contrast, ALK1 inhibition resulted in Smad1/5/8 suppression and alleviated high-dose TGF-beta 1-induced NPC degeneration. Taken together, it was concluded that high-doses of TGF-beta 1 contributed to the degeneration of NPCs via the upregulation of ALK1 and Smad1/5/8.