补骨脂异黄酮对炎症小体的调控作用及机制初探

目的基于小鼠骨髓巨噬细胞系,建立NLRP3、NLRC4及AIM2炎症小体活化模型,探索补骨脂异黄酮对炎症小体的调控作用及初步机制。方法采用LPS联合ATP、Nigericin、Salmonella和Poly(dA∶dT)等刺激因子,分别构建NLRP3、NLRC4和AIM2炎症小体活化模型;Caspase-Glo~® 1 Inflammasome Assay检测caspase-1活性;免疫印迹法检测细胞培养上清中mature IL-1βp17、caspase-1 p20的蛋白水平,以及细胞裂解液中pro-caspase-1、pro-IL-1β、ASC、NLRP3等蛋白表达。结果补骨脂异黄酮可抑制LPS联合ATP、Nigericin、Salmonella和Poly(dA∶dT)诱导的pro-caspase-1自我剪切,抑制caspase-1介导的IL-1β产生,表明补骨脂异黄酮可抑制NLRP3、NLRC4及AIM2炎症小体活性。此外,研究发现补骨脂异黄酮对NLRP3炎症小体活性的抑制作用具有不可逆性,且不依赖于NF-κB通路。结论补骨脂异黄酮可通过抑制pro-caspase-1自我剪切活化,从而抑制NLRP3、NLRC4、AIM2炎症小体的激活,进一步抑制其介导的免疫炎症反应,本研究也在一定程度上为补骨脂异黄酮相关制剂治疗免疫炎症疾病提供依据。

Regulation and mechanism of corylin on inflammasomes

Aim To investigate the regulation effect and molecular mechanism of corylin on NLRP3,NLRC4,AIM2 inflammasomes with immortalized bone marrow-derived macrophages.Methods The effect of corylin on the NLRP3,NLRC4,AIM2 inflammasome activation was evaluated with LPS-induced ATP,nigericin,salmonella,poly(dA∶dT).Caspase-1 activity was determined by Caspase-Glo■1 Inflammasome Assay.Western blot was performed to observe the protein expression levels of mature IL-1β,caspase-1 p20 in the culture supernatants,pro-caspase-1,pro-IL-1β,ASC,NLRP3 in the cell lysates.Results Corylin blocked the self-slicing of pro-caspase-1 induced by ATP,nigericin,salmonella and poly(dA∶dT),then suppressed the secretion of mature IL-1βmediated by caspase-1,which showed that corylin inhibited the NLRP3,NLRC4,AIM2 inflammasomes activation.Moreover,corylin irreversibly attenuated the activation of NLRP3 inflammasome without affecting NF-κB signaling pathway.Conclusions Corylin inhibits the inflammasome activation of NLRP3,NLRC4,and AIM2,and further reduces its mediated immune inflammatory response.Meanwhile,it provides new ideas and strategies for the treatment of immune inflammatory diseases by using corylin-related preparations.