Effects of IGF-1 on IL-1β-induced apoptosis in rabbit nucleus pulposus cells in vitro.

Excessive apoptosis in intervertebral disc (IVD) cells is important in IVD degeneration. Interleukin (IL)-1β has been shown to induce apoptosis in these cells. However, whether insulin-like growth factor-1 (IGF-1) inhibits IL-1β-induced apoptosis in the nucleus pulposus remains unclear. The purpose of this study was to investigate the effects of IGF-1 on IL-1β-induced apoptosis in the nucleus pulposus. Cells isolated from the nucleus pulposus were grown in culture to a monolayer. These cells were identified using immuno-histochemistry for type II collagen and toluidine blue staining for glycosaminoglycans. Following exposure to IGF-1 or IL-1β, the cells were observed using light microscopy. Giemsa staining, TdT-mediated dUTP-biotin nick end-labeling (TUNEL) and flow cytometry (FCM) were used to detect the rate of early cell death, which served as an indicator of apoptosis. In the IL-1β group, a large number of these cells underwent apoptosis and demonstrated morphological changes associated with apoptosis. A small proportion of cells exposed to IGF-1 alone underwent apoptosis. No obvious signs of apoptosis were observed in the control group. TUNEL results revealed that the rate of apoptosis in the IGF-1 group was significantly reduced compared with that in the IL-1β group (P<0.01), confirmed using FCM. Compared with the control group, the apoptotic rate was also significantly increased in IL-1β-exposed cells (P<0.01). These findings strongly suggested that IGF-1 inhibits IL-1β-induced apoptosis in the nucleus pulposus.