Shikonin protects against lipopolysaccharide-induced inflammation and apoptosis in human nucleus pulposus cells through the nuclear factor-kappa B pathway

Shikonin significantly inhibited the inflammatory response and apoptosis of human primary NP cells, possibly through the NF-κB pathway.

Human primary nucleus pulposus (NP) cells cultured in vitro were used for the experiments. The effects of different concentrations of shikonin (1, 2, 4, 8, and 16 µM) on the activity of lipopolysaccharide (LPS)-induced NP cells were determined using the CCK-8 assay, and the appropriate drug concentration was determined. The experiment was divided into the control, LPS, and LPS + shikonin groups. ELISA and Western blot were used to detect the expression of the inflammatory factors tumor necrosis factor (TNF)-α and interleukin (IL)-1β. NP cell apoptosis was measured using Western blot and caspase 3 activity. Western blot and immunofluorescence assays were used to detect the protein expression of p-P65 and P65 and the nuclear translocation of P65.

To investigate the protective effect and mechanism of shikonin on human intervertebral disk degeneration.

The CCK-8 assay showed that shikonin had no cytotoxic effect on NP cells and increased the activity of LPS-induced NP cells, especially at a concentration of 4 μM. Shikonin reversed the expression of the inflammatory cytokines TNF-α and IL-1β and apoptosis-related molecules Bax, Bcl-2, and cleaved caspase 3 in LPS-induced NP cells. In addition, shikonin significantly decreased apoptosis and caspase-3 activity in LPS-induced NP cells. Furthermore, shikonin treatment significantly inhibited the expression of p-P65 and nuclear translocation of P65, and nuclear factor-kappa B (NF-κB) pathway inhibitor Pyrrolidinedithiocarbamate ammonium (PDTC) significantly enhanced the anti-inflammatory and antiapoptotic effects of shikonin in LPS-induced NP cells.