kukoamine-a and Intervertebral-Disc-Degeneration

Intervertebral disc degeneration (IDD) is the leading cause of back, neck, and radicular pain. This study aims to look at the roles of Kukoamine A (KuA) in nucleus pulposus cells (NPCs) of IDD and its related potential mechanisms. Cell viability of NPCs in the control, lipopolysaccharide (LPS) and LPS+KuA groups was firstly detected by cell counting kit (CCK)-8. Meanwhile, the protein expression of collagen II in LPS-induced NPCs was measured by western blot. Then, the experiments following the treatment of KuA in LPS-induced NPCs included cell proliferation assessment by 5-ethynyl-2'-deoxyuridine (EdU) kit, cell apoptosis and extracellular matrix degradation (ECM) analysis by Terminal dUTP nick-end labeling (TUNEL) and western blot, the detection of inflammatory cytokines by western blot and enzyme-linked immunosorbent assay (ELISA), P13K/Akt pathway-related protein levels analysis by western blot. Finally, after the addition of P13K/Akt pathway inhibitor LY294002, cell apoptosis, ECM and inflammation in KuA-treated NPCs induced by LPS were again examined by the same methods. Results indicated that KuA prevented loss of cell viability and attenuated the apoptosis, ECM, and inflammation in LPS-induced NPCs. Furthermore, western blot experiment verified the activation of KuA on P13K/Akt pathway in LPS-induced NPCs. However, inhibition of P13K/Akt pathway reversed the roles of KuA in LPS-induced NPCs. Thus, KuA attenuates LPS-induced apoptosis, ECM and inflammation in LPS-induced NPCs by activating the P13K/Akt pathway.

Topics: Apoptosis; Cells, Cultured; Extracellular Matrix; Humans; Inflammation; Intervertebral Disc Degeneration; Lipopolysaccharides; Nucleus Pulposus; Proto-Oncogene Proteins c-akt; Spermine