Jang Ah-Ra, Lee Ha-Nul, Hong Jung Joo, Kim Young-Min, Park Jong-Hwan
Laboratory of Animal Medicine, College of Veterinary Medicine and Animal Medical Institute, Chonnam National University, Gwangju 61186, Republic of Korea.
NodCure, Inc., Gwangju 61186, Republic of Korea.
Exp Ther Med. 2023 Feb 7;25(3):128. doi: 10.3892/etm.2023.11827. eCollection 2023 Mar.
() is used in traditional East Asian medicine for the treatment of various diseases, including inflammatory disease. However, it has remained unclear whether extracts of inhibit inflammasome activation in macrophages. The present study assessed the inhibitory effect of an ethanol extract of (CZE) on the activation of the inflammasome in macrophages and the underlying mechanism. Bone marrow-derived macrophages were obtained from wild-type C57BL/6 mice. The release of IL-1β and lactate dehydrogenase in response to nucleotide-binding oligomerization domain-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome activators, such as ATP, nigericin and monosodium urate (MSU) crystals, was significantly decreased by CZE in lipopolysaccharide (LPS)-primed BMDMs. Western blotting revealed that CZE inhibited ATP-induced caspase-1 cleavage and IL-1β maturation. To investigate whether CZE inhibits the priming step of the NLRP3 inflammasome, we confirmed the role of CZE at the gene level using RT-qPCR. CZE also downregulated the gene expression of NLRP3 and pro-IL-1β as well as NF-κB activation in BMDMs in response to LPS. Apoptosis-associated speck-like protein containing a caspase-recruitment domain (CARD) oligomerization and speck formation by NLRP3 inflammasome activators were suppressed by CZE. By contrast, CZE did not affect NLR family CARD domain-containing protein 4 or absent in melanoma 2 inflammasome activation in response to and poly(dA:dT) in LPS-primed BMDMs, respectively. The results revealed that three key components of CZE, namely linarin, 3,5-dicaffeoylquinic acid and chlorogenic acid, decreased IL-1β secretion in response to ATP, nigericin and MSU. These findings suggest that CZE effectively inhibited activation of the NLRP3 inflammasome.
（）在传统东亚医学中用于治疗包括炎症性疾病在内的各种疾病。然而，其提取物是否能抑制巨噬细胞中的炎性小体激活仍不清楚。本研究评估了（某植物名，文中未明确给出具体中文名）乙醇提取物（CZE）对巨噬细胞中炎性小体激活的抑制作用及其潜在机制。从野生型C57BL/6小鼠中获取骨髓来源的巨噬细胞。在脂多糖（LPS）预处理的骨髓来源巨噬细胞（BMDMs）中，CZE显著降低了对核苷酸结合寡聚化结构域样受体（NLR）家族含pyrin结构域3（NLRP3）炎性小体激活剂（如ATP、尼日利亚菌素和尿酸单钠（MSU）晶体）的反应中白细胞介素-1β（IL-1β）和乳酸脱氢酶的释放。蛋白质免疫印迹法显示CZE抑制了ATP诱导的半胱天冬酶-1裂解和IL-1β成熟。为了研究CZE是否抑制NLRP3炎性小体的启动步骤，我们使用逆转录定量聚合酶链反应（RT-qPCR）在基因水平上证实了CZE的作用。CZE还下调了LPS刺激的BMDMs中NLRP3和前体IL-1β的基因表达以及核因子κB（NF-κB）的激活。CZE抑制了NLRP3炎性小体激活剂诱导的含半胱天冬酶招募结构域（CARD）的凋亡相关斑点样蛋白的寡聚化和斑点形成。相比之下，CZE分别对LPS预处理的BMDMs中对（某物质，文中未明确给出具体中文名）和聚（dA:dT）的反应中NLR家族含CARD结构域蛋白4或黑色素瘤缺失2炎性小体激活没有影响。结果显示，CZE的三个关键成分，即香叶木素、3,5-二咖啡酰奎宁酸和绿原酸，降低了对ATP、尼日利亚菌素和MSU的反应中IL-1β的分泌。这些发现表明CZE有效地抑制了NLRP3炎性小体的激活。
