Effect Of Zinc Finger Protein A20 And NF-κB In The Rabbit Lumbar Disc Degeneration Animal Modle

Objective:To observe the expression of zinc finger protein A20,NF-κB in normal and degenerated rabbit lumbar intervertebral disc,the nucleus pulposus cells were cultured in vitro and further stimulated with endotoxin(LPS)to observe the expression and relationship of zinc finger protein A20,NF-κB and related inflammatory factors.Methods:To obtain the normal and degenerative intervertebral disc tissue in rabbit lumbar disc degeneration animal models,HE staining observe the morphological changes of nucleus pulposus and annulus fibrosus,immunohistochemistry was used to detect the expression of A20、NF-κB/p65 and COL-Ⅱ.The nucleus pulposus cells were isolated and cultured in vitro,then divided into normal group,degenerative group,LPS stimulation group and NF-κB inhibition group,Real-time PCR was employed to analyze the expression of A20、IL-1β、TNF-α、NF-κB、COL-Ⅱ,Western Blot was used to observe the A20 protein,p65 and COL-Ⅱexpression changed in the four groups,and TNF-α,IL-1β in cell supernatant was determined by ELISA.Results:Histological observation revealed the number of nucleus pulposus cells was significantly decreased,aggregation occurs in the degenerative group.COL-Ⅱ was obvious lower and A20,p65 was significantly higher than normal group by immunohistochemical staining.Cell experiments found that compared with the normal group,A20、TNF-α、IL-1β、p65 expression was significantly increased and COL-Ⅱdecreased in the mRNA and protein levels in degenerative group.above indexes Change more significant in LPS stimulation group than in degenerative group.after addition of NF-κB inhibitor,The above index expression instead,and have significant statistical significance with the LPS stimulation group(p<0.05).Conclusion:When the rabbit lumbar disc degeneration occurs,the expression of zinc finger protein A20 and the activity of NF-κB increased.After adding LPS,the expression of zinc finger protein A20,NF-κB and its related inflammatory factors was further increased,and zinc finger protein A20 and NF-κB was positively correlated.It suggested that the nucleus pulposus cell inflammatory response is closely related to the development of rabbit disc degeneration,which A20 may play an important role.