PMID- 31479750 OWN - NLM STAT- MEDLINE DCOM- 20200624 LR - 20200624 IS - 1096-1186 (Electronic) IS - 1043-6618 (Linking) VI - 149 DP - 2019 Nov TI - Identification of an IKKbeta inhibitor for inhibition of inflammation in vivo and in vitro. PG - 104440 LID - S1043-6618(19)30781-9 [pii] LID - 10.1016/j.phrs.2019.104440 [doi] AB - Targeting on the IKKbeta to discover anti-inflammatory drugs has been launched for ten years, due to its predominant role in canonical NF-kappaB signaling. In the current study, we identified a novel IKKbeta inhibitor, ellipticine (ELL), an alkaloid isolated from Ochrosia elliptica and Rauvolfia sandwicensis. We found that ELL reduced the secretion and mRNA expression of TNF-alpha and IL-6 and decreased the protein expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in bone marrow derived macrophages (BMDMs) stimulated with LPS. In coincided with the results, ELL suppressed PGE2 and NO production in BMDMs. Underlying mechanistic study showed that ELL inhibited IkappaBalpha phosphorylation and degradation as well as NF-kappaB nuclear translocation, which was attributed to suppression of IKKalpha/beta activation. Furthermore, kinase assay and binding assay results indicated that ELL inhibited IKKbeta activity via directly binding to IKKbeta and in turn resulted in suppression of NF-kappaB signaling. To identify the binding sites of ELL on IKKbeta, IKKbeta(C46A) plasmid was prepared and the kinase assay was performed. The results demonstrated that the inhibitory effect of ELL on IKKbeta activity was impaired in the mutation, implying that anti-inflammatory effect of ELL was partially attributed to binding on cysteine 46. Furthermore, ELL up-regulated LC3 II expression and reduced p62 expression, suggesting that autophagy induction contributed to the anti-inflammatory effect of ELL as well. In coincided with the in vitro results, ELL increased the survival and antagonized the hypothermia in the mice with LPS-induced septic shock. Consistently, ELL reduced TNF-alpha and IL-6 production in the serum of the mice treated with LPS. Collectively, our study provides evidence that ELL is an IKKbeta inhibitor and has potential to be developed as a lead compound for treatment inflammatory diseases in the future. CI - Copyright (c) 2019 Elsevier Ltd. All rights reserved. FAU - Chen, Qi AU - Chen Q AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China; Nursing Department, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, 646000, China. FAU - Liu, Juan AU - Liu J AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Zhuang, Yuxin AU - Zhuang Y AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Bai, Li-Ping AU - Bai LP AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Yuan, Qing AU - Yuan Q AD - Department of Basic Medicine, Southwest Medical University, Luzhou, Sichuan Province, 646000, China. FAU - Zheng, Silin AU - Zheng S AD - Nursing Department, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, 646000, China. FAU - Liao, Kangsheng AU - Liao K AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Khan, Md Asaduzzaman AU - Khan MA AD - Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Southwest Medical University, Room 3-319, Zhongshan Road, Luzhou, Sichuan, 646000, China. FAU - Wu, Qibiao AU - Wu Q AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Luo, Cheng AU - Luo C AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Liu, Liang AU - Liu L AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. FAU - Wang, Hui AU - Wang H AD - School of Biological Medicine, Beijing City University, Beijing, 100084, China. Electronic address: vera_wh@126.com. FAU - Li, Ting AU - Li T AD - State Key Laboratory of Quality Research in Chinese Medicine/Macau Institute for Applied Research in Medicine and Health, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau, China. Electronic address: tli@must.edu.mo. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190831 PL - Netherlands TA - Pharmacol Res JT - Pharmacological research JID - 8907422 RN - 0 (Anti-Inflammatory Agents) RN - 0 (Ellipticines) RN - 117VLW7484 (ellipticine) RN - EC 2.7.11.10 (I-kappa B Kinase) SB - IM MH - Animals MH - Anti-Inflammatory Agents/chemistry/pharmacology/*therapeutic use MH - Cells, Cultured MH - Drug Discovery MH - Ellipticines/chemistry/pharmacology/*therapeutic use MH - Female MH - Humans MH - I-kappa B Kinase/*antagonists & inhibitors/immunology MH - Inflammation/*drug therapy/immunology MH - Mice MH - Ochrosia/chemistry MH - Shock, Septic/*drug therapy/immunology OTO - NOTNLM OT - IKK OT - Inflammation OT - Inhibitor EDAT- 2019/09/04 06:00 MHDA- 2020/06/25 06:00 CRDT- 2019/09/04 06:00 PHST- 2019/05/05 00:00 [received] PHST- 2019/08/19 00:00 [revised] PHST- 2019/08/29 00:00 [accepted] PHST- 2019/09/04 06:00 [pubmed] PHST- 2020/06/25 06:00 [medline] PHST- 2019/09/04 06:00 [entrez] AID - S1043-6618(19)30781-9 [pii] AID - 10.1016/j.phrs.2019.104440 [doi] PST - ppublish SO - Pharmacol Res. 2019 Nov;149:104440. doi: 10.1016/j.phrs.2019.104440. Epub 2019 Aug 31.