Effect of Inotodiol on LPS-induced Injury of RAW264.7 Cells

Abstract

To explore the effect of Inotolinol (INO) on the inflammatory response of LPS-induced mouse mononuclear macrophages (RAW264.7). The experiment has investigated in vitro the effects of INO on mouse mononuclear macrophages (RAW264.7) induced by LPS. To do this, the following methods were adopted: The viability of RAW264.7 cells was detected by CCK-8 assay. Cell apoptosis was examined by Hoechst33342 and PI. The secretion of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-18 (IL-18) and the content of nitric oxide (NO) in cells were measured by ELISA and Griess. The production of reactive oxygen species (ROS), the content of malondialdehyde (MDA) and hydrogen peroxide (H2O2), and the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) in cells were detected by fluorescence probe and kit. The results showed that 20 μmol/L INO could inhibit the decline of RAW264.7 cell viability induced by LPS, prevent cell apoptosis, and significantly inhibit the secretion of IL-6, IL-1β, TNF-α, IL-18 and NO (P<0.01), could significantly reduce the contents of MDA and H2O2 in cells (P<0.01), and could significantly increase the activities of SOD, CAT and GSH in cells (P<0.01). The above results indicate that INO can effectively inhibit the production of pro-inflammatory factors and superoxides induced by LPS in RAW264.7 cells, improve the anti-inflammatory and anti-oxidative abilities of cells, and thus protect cells from damage.

Keywords

• inotodiol
• lps
• raw264.7 cells
• the inflammatory response
• oxidative stress