目的 探讨雷公藤红素对小鼠巨噬细胞RAW264.7焦亡的影响及相关机制。方法 采用MTT法测定细胞增殖；吖啶橙（AO）/溴化乙啶（EB）、Hoechst/PI荧光染色观察细胞焦亡形态变化；ELISA法检测细胞上清液中白细胞介素-1β（IL-1β）分泌量；Western blotting法检测Caspase-1蛋白表达水平；Caspase-1活性检测试剂盒检测细胞内Caspase-1酶活性。结果 经MTT法检测发现，雷公藤红素在50 nmol/L以下对巨噬细胞RAW264.7增殖活性无显著影响；AO/EB、Hoechst/PI染色显示雷公藤红素能够改善脂多糖（LPS）与三磷酸腺苷（ATP）诱导的细胞焦亡；ELISA结果显示，雷公藤红素能够抑制IL-1β的分泌（P<0.05），且具有一定的浓度依赖性；Western blotting检测发现cleaved-Caspase-1蛋白表达量降低，同时雷公藤红素能抑制Caspase-1的酶活性。结论 雷公藤红素抑制LPS与ATP诱导的细胞焦亡，可能与抑制IL-1β的分泌，抑制Caspase-1的活化有密切关系。

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.

国家自然科学基金项目（81503339）；山东省博士基金资助项目（BS2014YY049）；烟台市科技计划资助项目（2014ZH092）；科研启动基金资助项目（BY2013KYQD09）