摘要
目的以LPS致敏的小鼠巨噬细胞J774A.1作为炎症细胞模型,研究灯盏花乙素对ATP诱导的炎症小体活化和细胞焦亡的影响及其机制。方法利用碘化丙锭(PI)染色法检测LPS+ATP诱导的小鼠J774A.1巨噬细胞发生细胞焦亡的情况;免疫印迹法检测细胞裂解液和上清中IL-1β、caspase-1、HMGB1等蛋白的表达水平;基于微珠的免疫测定法(CBA)检测细胞上清中IL-1β的分泌水平。结果 ATP能够明显诱导LPS致敏的J774A.1巨噬细胞中caspase-1活化、成熟IL-1β(17 ku)和HMGB1释放至培养上清中,并诱导细胞焦亡;而灯盏花乙素预处理能够剂量依赖性地抑制ATP诱导的caspase-1活化以及成熟IL-1β、HMGB1的释放,并抑制细胞焦亡;同时,经腺苷酸环化酶抑制剂MDL12330A和蛋白激酶A(PKA)抑制剂H89处理,可以逆转灯盏花乙素对ATP诱导的细胞焦亡的抑制作用。结论灯盏花乙素通过调节PKA活性,抑制NLRP3炎症小体的活化与细胞焦亡,从而发挥抗炎作用。
Aim To explore the influences of scutellarin on ATP-induced NLRP3 inflammasome activation and pyroptosis,using LPS-primed murine macrophages J774 A. 1 as an inflammatory cell model,and to explore the underlying mechanism. Methods The effects of scutellarin on ATP-induced pyroptosis in murine J774 A. 1 macrophages were analyzed by propidium iodide( PI) staining assay. The levels of IL-1β,caspase-1 and HMGB1 in cell lysates and culture supernatants were analysed using Western blot. The levels of IL-1β in cell culture supernatants were measured by cytometric beads array( CBA). Results ATP significantly induced caspase-1 activation and mature IL-1β and HMGB1 release into the culture supernatants in LPS-primed murine J774 A. 1 macrophages, and induced pyroptosis. Scutellarin treatment dose-dependently inhibited ATP-induced caspase-1 activation,mature IL-1β and HMGB1 release,and pyroptosis. Notably,scutellarin 's inhibitory effects on ATP-induced pyroptosis were markedly reversed by the adenylate cyclase inhibitor MDL12330 A and selective protein kinase A( PKA) inhibitor H89. Conclusion Scutellarin inhibits NLRP3 inflammasome activation and pyroptosis by modulating the PKA activity in macrophages,thereby exhibiting anti-inflammatory activities.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第2期174-180,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81773965
81673664)
