| Objective:To observe the relationship between nuclear factor kapp B(NF-κB)activation and cell apoptosis,before and after LPS stimulation of human degene-rateve nucleus pulposus cells.Methods:Degenerative human nucleus pulposus cells were cultured in vitro.Useing CCK-8 to observe the proliferative effect of LPS on the nucleus pulposus cells in vitro,in the concentration of 100,200,500 and 1 000 μg/mL and choose the most apropriate concentration.The experiment was divided into 3 groups: blank control group,LPS(500 μg/mL)groups,and NF-κB inhibitor(BAY11-7082)plus LPS(500 μg/mL)group,Annexin V-FITC/PI flow cytometry and Hoechest33258 staining was used to analyze apoptosis.The expression of cleaved caspase 3,PARP,P65,P-P65 proteins were detected by Western blot respectively.Results:The results of CCK-8 assay showed that when LPS was 500 μg/mL,the cell vitality was declined obviously.Compared with the blank control group,cell apoptosis rate of the LPS group is increasing(p<0.05),and the expression of P-P65,cleaved caspase 3,cleaved PARP were also higher(p< 0.05).Compared with the LPS group,cell apoptosis rate of the NF-κB inhibitor plus LPS group is significantly lower(p<0.05)and the expression of P-P65,cleaved caspase 3,cleaved PARP were also lower(p< 0.05).Conclusions:NF-κB signaling pathway may be associated with the nucleus pulposus cell apoptosis in disc degeneration,so it was worth further study. |
