Effect of celastrol on pyroptosis of macrophages RAW264.7 / 中草药

ABSTRACT

Objective: To investigate the effects and underlying mechanism of celastrol on pyroptosis in macrophage RAW264.7. Methods: Cell viability was determined by using MTT assay. Effects of celastrol on pyroptosis of macrophages were observed through AO/EB and Hoechst/PI fluorescence staining. The secretions of IL-1β in the culture supernatant were analyzed by ELISA. The protein levels of Caspase-1 was assayed by Western blotting. And Caspase-1 activity was detected with Caspase-1 activity detection kit. Results: Based on the MTT assay, the concentrations under 50 nmol/L of celastrol used were not toxic to the macrophages. PI or EB uptake induced by lipopolysaccharide (LPS) and ATP were significantly reduced by celastrol. Pretreatment with celastrol greatly reduced the secretion of IL-1β in a dose-dependent manner, and inhibited the up-regulation of cleaved-Caspase-1. Additionally, celastrol could inhibit the activation of Caspase-1 in macrophages. Conclusion: Celastrol significantly reduced pyroptosis induced by LPS and ATP through inhibition of IL-1β secretion and Caspase-1 activation.