Changes In The Expression Of Zinc Finger Protein A20 And Inflammatory Factors In Nucleus Pulposus Cells During Intervertebral Disc Degeneration

Objective: To detecte the expression of zinc finger protein A20 in nucleus pulposus of human degenerative intervertebral disc; and to explore the expression changes of zinc finger protein A20 in vitro with LPS induced prenatal mouse intervertebral disc nucleus pulposus cell.Methods: It was observed the degenerative situation of human nucleus pulposus by Alcian blue and Immunohistochemical observated the expression of zinc finger protein A20 in human intervertebral disc nucleus pulposus cells. Experimental group was prenatal mouse intervertebral disc nucleus pulposus cell by LPS-stimulated, NF- kappa B inhibition group was prenatal mouse intervertebral disc nucleus pulposus cell by LPS-stimulated and BAY 11-7082(NF-κB inhibitor),The control group was prenatal mouse intervertebral disc nucleus pulposus cell by added the same amount of PBS. MTT was used to observe the cytotoxicity of 0.01μg/ml concentrations of LPS on prenatal mouse intervertebral disc nucleus pulposus cell. With Real- time PCR observing prenatal mouse intervertebral disc nucleus pulposus cell the expression of zinc finger protein of A20、TNF-α、IL-1β、ADAMST-4、ADAMST-5、MMP-13 changed; Using Western Blot observing zinc finger protein of A20 protein, p65, p-p65 expression changed in prenatal mouse intervertebral disc nucleus pulposus cell.Results: Observed human degenerative intervertebral disc nucleus pulposus cell decrease, aggregation occurs by Alcian blue staining. Observed in human degenerative intervertebral disc nucleus pulposus cell A20 was significantly higher than the normal group by immunohistochemical staining. Prenatal mouse intervertebral disc nucleus pulposus cell induced by LPS, with Real-time PCR it was showed that the expression of A20, TNF-α, IL-1β, ADAMST-4, ADAMST-5, MMP-13 and mRNA were higher than NF- kappa B inhibition group and the control group in the experimental group(P<0.05), by Western Blot it was found in the experimental group of A20, TNF-α, IL-1β, ADAMST-4, ADAMST-5, MMP-13, p65 and p-p65 protein were significantly higher than those in NF- kappa B inhibition group and the control group(P <0.05).Conclusion: Increased expression of A20 in intervertebral disc degeneration. LPS stimulation, in the prenatal mouse intervertebral disc nucleus pulposus cell A20 and inflammatory factor expression increased, prior to and after the inhibition of NF- B activity were significant, suggest that cells of the nucleus pulposus inflammatory reaction and intervertebral disc degeneration may be associated and A20 in which may play an important role.