Abstract:

Based on bioinformatics, this study validates the mechanism of action of Haitongpi-Tougucao (compound Haitongpi) in inhibiting lipopolysaccharide (LPS)-induced ferroptosis in rat inflammatory chondrocytes. Bioinformatics tools were used to predict the mechanism of action of ferroptosis in osteoarthritis and identify pathways for validation. The key techniques used were as follows: the detection of ferrous ion content and reduced glutathione (GSH) content using relevant kits; the detection of cell viability and the levels of related cytokines IL-1β, IL-6, and TNF-α using the enzyme-linked immunosorbent assay (ELISA) after dosing; and the use of protein immunoblotting (western blot, WB) to detect the protein expression levels of NLRP3, Caspase-1, ASC, and GPX4, a gene that inhibits ferroptosis, related to the NLRP3 inflammasome pathway in each group. The results revealed that the ferrous ion content was significantly decreased,while the GSH content was significantly increased; the ELISA experiment showed that the levels of inflammatory factors IL-1β, IL-6, and TNF-α were decreased in each group administered with the drug compared with those in the model group; the WB results showed that the expression levels of NLRP3, Caspase-1, and ASC proteins were significantly decreased and GPX4 protein expression levels were significantly increased in each group administered with a specific dosage of the drug compared with those in the model group. Therefore, the compound Haitongpi can intervene in the ferroptosis of inflammatory chondrocytes by mediating the NLRP3 inflammasome pathway, thereby achieving the purpose of osteoarthritis treatment.

Key words: osteoarthritis, ferroptosis, Haitongpi-tougucao, bioinformatics, NLRP3 inflammasome pathway