PMID- 35597027 OWN - NLM STAT- MEDLINE DCOM- 20220614 LR - 20220614 IS - 1618-095X (Electronic) IS - 0944-7113 (Linking) VI - 102 DP - 2022 Jul 20 TI - The ethanolic extract of Artemisia anomala exerts anti-inflammatory effects via inhibition of NLRP3 inflammasome. PG - 154163 LID - S0944-7113(22)00241-0 [pii] LID - 10.1016/j.phymed.2022.154163 [doi] AB - BACKGROUND: Artemisia anomala S. Moore (Compositae), known as "Nan-Liu-Ji-Nu" in traditional Chinese medicine (TCM), has been used to treat many inflammatory diseases, including enteritis, acute icteric hepatitis, rheumatism, toothache, tonsillitis, and chronic bronchitis, for centuries. Our preliminary studies have demonstrated that the ethanolic extract of A. anomala (EAA) might be with the potential of inhibiting the activation of the NLRP3 inflammasome. However, the anti-inflammatory activity of EAA based on NLRP3 inflammasome inhibition is still unclear. PURPOSE: This work aimed to elucidate the anti-inflammatory mechanism of EAA by inhibiting NLRP3 inflammasome activation. METHODS: Lipopolysaccharide (LPS)-primed bone marrow-derived macrophages (BMDMs) were used to evaluate the inhibitory effects on NLRP3 inflammasome activation. The level of IL-1beta was determined by ELISA. The expression levels of IL-1beta, caspase-1, NLRP3, and ASC were assayed using western blot analysis. ASC oligomerization and speck formation were detected by immunofluorescence microscopy. The measurements of intracellular chloride and potassium were conducted using N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) probe assay and inductively coupled plasma-optical emission spectrometry (ICP-OES), respectively. Mitochondrial reactive oxygen species (mtROS) were examined using the MitoSOX method. Acridine orange (AO) staining was used to detect the permeability of the lysosomal membrane. A DSS-induced ulcerative colitis model was established to evaluate the anti-inflammatory effects of EAA in vivo. Finally, high-performance liquid chromatography (HPLC) was employed to identify and quantify the major constituents of EAA. RESULTS: In BMDMs, EAA significantly inhibited the release of IL-1beta induced by LPS. The mechanistic study revealed that EAA inhibited NLRP3 inflammasome activation by blocking the oligomerization of ASC and suppressed the LPS-induced priming step. Furthermore, EAA protected lysosomes by inhibiting the TAK1-JNK pathway, thereby inhibiting the assembly of downstream NLRP3 inflammasome and the production of IL-1beta. In addition, EAA exerted potent protective effects in an ulcerative colitis model by decreasing the content of colonic IL-1beta and alleviating the process of ulcerative colitis. HPLC analysis identified eight main components of EAA, including isofraxidin (1), quercetin-7-O-beta-D-glucopyranoside (2), apigenin-7-O-beta-D-glucopyranoside (3), 7-methoxycoumarin (4), quercetin (5), luteolin (6), kaempferol (7), and eupatorin (8), Of these compounds, quercetin and kaempferol were found to be the most potent ingredients. CONCLUSION: These findings collectively reveal that EAA exerts anti-inflammatory effects by both suppressing the NLRP3 priming step and protecting lysosomes to inhibit NLRP3 inflammasome activation, suggesting that this traditional herbal medicine might be used to treat NLRP3-driven inflammatory diseases. CI - Copyright (c) 2022. Published by Elsevier GmbH. FAU - Hong, Feng AU - Hong F AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Zhao, Min AU - Zhao M AD - Laboratory of Metabolomics and Drug-induced Liver Injury, Frontiers Science Center for Disease-related Molecular Network, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Xue, Lin-Lin AU - Xue LL AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Ma, Xu AU - Ma X AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Liu, Ling AU - Liu L AD - School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, PR China. FAU - Cai, Xiao-Ying AU - Cai XY AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Zhang, Rui-Jia AU - Zhang RJ AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Li, Na AU - Li N AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Wang, Lun AU - Wang L AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. FAU - Ni, Heng-Fan AU - Ni HF AD - School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, PR China. FAU - Wu, Wen-Shuang AU - Wu WS AD - Department of Thyroid Surgery, West China Hospital, Sichuan University, Chengdu 610041, PR China. Electronic address: wenshuang_wu@163.com. FAU - Ye, Hao-Yu AU - Ye HY AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. Electronic address: haoyu_ye@scu.edu.cn. FAU - Chen, Li-Juan AU - Chen LJ AD - Laboratory of Natural Product Drugs, State Key Laboratory of Biotherapy and Cancer Center, West China Medical School, West China Hospital, Sichuan University, Chengdu 610041, PR China. LA - eng PT - Journal Article DEP - 20220510 PL - Germany TA - Phytomedicine JT - Phytomedicine : international journal of phytotherapy and phytopharmacology JID - 9438794 RN - 0 (Anti-Inflammatory Agents) RN - 0 (Inflammasomes) RN - 0 (Interleukin-1beta) RN - 0 (Kaempferols) RN - 0 (Lipopolysaccharides) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (Plant Extracts) RN - 9IKM0I5T1E (Quercetin) RN - EC 3.4.22.36 (Caspase 1) SB - IM MH - Anti-Inflammatory Agents/pharmacology MH - *Artemisia MH - Caspase 1/metabolism MH - *Colitis, Ulcerative MH - Inflammasomes MH - Interleukin-1beta/metabolism MH - Kaempferols MH - Lipopolysaccharides/pharmacology MH - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism MH - Plant Extracts/pharmacology MH - Quercetin OTO - NOTNLM OT - Anti-inflammatory activity OT - Artemisia anomala OT - EAA OT - NF-kappaB OT - NLRP3 inflammasome EDAT- 2022/05/22 06:00 MHDA- 2022/06/15 06:00 CRDT- 2022/05/21 18:16 PHST- 2021/03/14 00:00 [received] PHST- 2022/04/11 00:00 [revised] PHST- 2022/05/08 00:00 [accepted] PHST- 2022/05/22 06:00 [pubmed] PHST- 2022/06/15 06:00 [medline] PHST- 2022/05/21 18:16 [entrez] AID - S0944-7113(22)00241-0 [pii] AID - 10.1016/j.phymed.2022.154163 [doi] PST - ppublish SO - Phytomedicine. 2022 Jul 20;102:154163. doi: 10.1016/j.phymed.2022.154163. Epub 2022 May 10.