Abstract:
OBJECTIVE To observe the effect of Wenjing Tongluo Decoction on IL-1β-induced inflammation of articular chondrocytes and explore its mechanism.
METHODS Primary chondrocytes were extracted from knee cartilage of mice and identified by toluidine blue staining. The chondrocyte inflammation model was induced by IL-1β. Lentiviral RNAi-HMGB1 was transfected into chondrocytes to interfere the expression of HMGB1. Then the cells were intervened with different concentrations of Wenjing Tongluo Decoction containing serum. Western Blot was used to analyze the protein expression of HMGB1 and NF-κB signaling pathway. The expression of HMGB1, NF-κB1, NF-κB2 mRNA was detected by qPCR. ELISA was applied to detect the expression of inflammatory factors IL-6, PGE-2 and TNF-α in cell supernatants.
RESULTS Compared with normal cultured cells, the expression of HMGB1 increased significantly (P < 0.01) in the IL-1β-induced chondrocyte inflammatory model, and the relative expression of p-IκBα/IκBα and p-P65/P65 proteins and the expression of NF-κB1 and NF-κB2 mRNA were increased (P < 0.01). Compared with the model group, lentiviral RNAi-HMGB1 transfection could significantly downregulate the expression of HMGB1 (P < 0.01), and inhibit the upregulation of the relative expression of p-IκBα/IκBα (P < 0.01) and p-P65/P65 protein and NF-κB1 and NF-κB2 mRNA expression (P < 0.05). Compared with the model group, different concentrations of Wenjing Tongluo Decoction showed inhibitory effects on HMGB1 and NF-κB signaling pathways, demonstrated by the decrease in the protein expression of HMGB1, p-IκBα/IκBα and p-P65/P65 (P < 0.05).
CONCLUSION Wenjing Tongluo Decoction can improve IL-1β-induced chondrocyte inflammation, and its mechanism may be related to inhibition of HMGB1-mediated activation of NF-κB signaling pathway.