Kukoamine A attenuates lipopolysaccharide-induced apoptosis, extracellular matrix degradation, and inflammation in nucleus pulposus cells by activating the P13K/Akt pathway.

Abstract

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.

• Authors: Dan Wang, Hao Qu, Hui Kang, Feng Xu, Wei Huang, Xianhua Cai
• Journal: Bioengineered (Bioengineered) Vol. 13 Issue 4 Pg. 8772-8784 (04 2022) ISSN: 2165-5987 [Electronic] United States
• PMID: 35333664 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Lipopolysaccharides
• kukoamine A
• Spermine
• Proto-Oncogene Proteins c-akt

Topics

• Apoptosis
• Cells, Cultured
• Extracellular Matrix (metabolism)
• Humans
• Inflammation (metabolism)
• Intervertebral Disc Degeneration
• Lipopolysaccharides (metabolism)
• Nucleus Pulposus (metabolism)
• Proto-Oncogene Proteins c-akt (metabolism)
• Spermine (analogs & derivatives)