Based On HMGB1/RAGE/NF-κB Pathway To Explore The Anti-inflammatory Molecular Mechanism Of Triptolide

Objective: The purpose of this study was to explore the effect of triptolide(TP)on the growth of mouse peritoneal macrophage RAW264.7 cells through a series of vitro experiments based on HMGB1/RAGE/NF-κB pathway.Methods: In this project,RAW264.7 cells were induced by lipopolysaccharide(LPS)at a dose of 0.5 μg/m L for 8 h to establish an inflammation model.CCK8 assay was used to screen the optimal concentration of TP,which was 25 n M.The cells were divided into 3 groups as follow: control group,LPS treated group,LPS +TP treated group.After 8 hours,the supernatant of cell culture medium was used for ELISA;protein extraction for western blot,RNA extraction for RT-PCR.The secretion of the inflammatory cytokines including tumor necrosis factor(TNF-α),interleukin-1β(IL-1β)and interleukin-10(IL-10)were detected by ELISA.Western blot assay was used to detect the protein expression levels of HMGB1,RAGE and NF-κB.RT-PCR was performed to detect the m RNA expression of HMGB1,RAGE and NF-κB.Results: Compared with the LPS treated group,content of IL-1β in the LPS + TP treated group was reduced significantly.There was a significantly statistical difference between the two groups.Furthermore,TP significantly reduced the protein and m RNA expression of HMGB1,RAGE and NF-κB.The differences were statistically significant.Conclusions: The results in vitro cell experiments showed that TP inhibited the production of inflammatory factors in RAW264.7 cells induced by LPS,and the mechanism may be related to the regulation of the HMGB1/RAGE/NF-κB signaling pathway.