目的 探讨隐丹参酮在丹参抑制NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）炎症小体活化中的作用。方法 采用骨髓源性巨噬细胞（bone marrow derived macrophage，BMDMs）建立NLRP3炎症小体活化模型，评价丹参提取物对NLRP3炎症小体活化的影响。利用目标成分敲出/敲入评价隐丹参酮在丹参抑制NLRP3炎症小体活化中的作用，采用薄层色谱法“敲出”隐丹参酮得到目标成分样品（隐丹参酮样品）及阴性样品（丹参提取物中敲出隐丹参酮的样品），并利用NLRP3炎症小体活化模型，比较其对NLRP3炎症小体活化的抑制作用，通过ELISA和Western blotting检测相关炎症因子的释放和蛋白的表达；利用“敲入”方法，在阴性样品中加入目标成分样品，并将之与丹参提取物进行比较，评价二者对NLRP3炎症小体活化的调控作用；同时利用质谱对目标成分样品、阴性样品进行检测。结果 丹参提取物对NLRP3炎症小体的活化有抑制作用；“敲出”隐丹参酮后丹参提取物对NLRP3炎症小体活化的抑制作用减弱，“敲入”后抑制作用回升。结论 隐丹参酮是丹参抑制NLRP3炎症小体活化的关键成分。

Objective To explore the effect of cryptotanshinone in inhibiting the activation of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome in Danshen (Salviae Miltiorrhizae Radix et Rhizoma). Methods Bone marrow derived macrophages (BMDMs) was used to establish an NLRP3 inflammasome activation model and the effect of Salviae Miltiorrhizae Radix et Rhizoma extract on NLRP3 inflammasome activation was evaluated. Target component knock-out/knock-in was used to evaluate the effect of cryptotanshinone in inhibiting NLRP3 inflammasome activation in Salviae Miltiorrhizae Radix et Rhizoma. Thin layer chromatography was used to knock-out cryptotanshinone to obtain target component samples (cryptotanshinone samples) and negative samples (samples of Salviae Miltiorrhizae Radix et Rhizoma extract with cryptotanshinone knock-out), and the NLRP3 inflammasome activation model was used to compare its inhibitory effect on NLRP3 inflammasome activation. ELISA and Western blotting were used to detect the release of relevant inflammatory factors and protein expressions; The knock-in method which added the target component sample to the negative sample was used and was compared with Salviae Miltiorrhizae Radix et Rhizoma extract to evaluate their regulatory effects on NLRP3 inflammasome activation; Mass spectrometry was simultaneously used to detect target component samples and negative samples. Results Salviae Miltiorrhizae Radix et Rhizoma extract had an inhibitory effect on the activation of NLRP3 inflammasome; The inhibitory effect of Salviae Miltiorrhizae Radix et Rhizoma extract on NLRP3 inflammasome activation was weakened after knocking-out cryptotanshinone, but the inhibitory effect was restored after knocking-in. Conclusion Cryptotanshinone is the key component in Salviae Miltiorrhizae Radix et Rhizoma’s inhibition of NLRP3 inflammasome activation.

R285.5

国家自然科学基金资助项目（82003984，82230118）；北京市自然科学基金资助项目（7232321，7214296）；国家中医药管理局中医药创新团队及人才支持计划项目（ZYYCXTD-C-202005）；国家中医药管理局高水平中医药重点学科建设项目（zyyzdxk-2023309）