Celastrol targets IRAKs to block Toll-like receptor 4-mediated nuclear factor-κB activation

OBJECTIVE: Celastrol has been established as a nuclear factor-κB(NF-κB) activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that int erleukin-1 receptor-associated kinases(IRA Ks) are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB(e.g., the inter leukin-1 receptor(IL-1R)/Toll- like receptor(TLR) superfamily).METHODS: The human hepatocellular cell line(Hep G2) treated with palmitic acid(PA) was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.RESULTS: The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the Hep G 2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.CONCLUSION: The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.