Effect of nerve growth factor and its transforming tyrosine kinase protein and low-affinity nerve growth factor receptors on apoptosis of notochordal cells.

PURPOSE

The disappearance of notochordal cells by apoptosis is thought to be the starting point of intervertebral disc degeneration. The aim of this study was to determine the apoptotic pathway of notochordal cells as well as the anti-apoptotic potential of caspase inhibitors.

METHODS

Rat notochordal cells were isolated, cultured, and placed in either 0 % (apoptosis-promoting condition) or 10 % (normal control) foetal bovine serum (FBS). We identified and quantified apoptotic cell deaths and caspase activities. In addition, we examined the cells for expression of nerve growth factor (NGF) and its two receptors--TrkA (survival signal) and p75 (apoptotic signal)--and downstream pathways. Finally, we analysed the degree of anti-apoptotic effects of caspase inhibitors on the cells.

RESULTS

The apoptotic rate and expressions of caspase-8 (extrinsic pathway), -9 (intrinsic pathway), and -3 (common executioner) of notochordal cells were increased in 0 % FBS compared with those in 10 % FBS. Expressions of NGF, p75 receptor and JNK downstream pathways were also increased in 0 % FBS. In contrast, expressions of the TrkA receptor and Akt and MAPK downstream pathways were decreased in 0 % FBS. Pancaspase, capase-9 and capase-8 inhibitors significantly reduced apoptotic cell death.

CONCLUSIONS

Our results suggest that notochordal cells undergo apoptosis through both the intrinsic and extrinsic pathways by activation of NGF, p75 receptor, and the JNK downstream pathway. We also found that apoptosis of notochordal cells can be attenuated by caspase inhibitors. Caspase inhibitors may play a therapeutic role in delaying the starting point of disc degeneration that is due to inappropriate or premature excessive apoptosis of notochordal cells.