PMID- 32272431 OWN - NLM STAT- MEDLINE DCOM- 20210126 LR - 20210126 IS - 1950-6007 (Electronic) IS - 0753-3322 (Linking) VI - 126 DP - 2020 Jun TI - Shikonin mitigates ovariectomy-induced bone loss and RANKL-induced osteoclastogenesis via TRAF6-mediated signaling pathways. PG - 110067 LID - S0753-3322(20)30258-4 [pii] LID - 10.1016/j.biopha.2020.110067 [doi] AB - BACKGROUND: Postmenopausal osteoporosis results from estrogen withdrawal and is characterized mainly by bone resorption. Shikonin is a bioactive constitute of Chinese traditional herb which plays a role in antimicrobial and antitumor activities. The study was designed to investigate the role of shikonin on postmenopausal osteoporosis and explore its underlying mechanisms. METHODS: Immunofluorescence staining was performed to evaluate the effects of shikonin on actin ring formation. The expression levels of the nuclear factor kappa-B (NF-kappaB) and mitogen-activated protein kinase (MAPK) pathway were determined by Western blot analysis. To determine whether shikonin influences the receptor activator of nuclear factor-kappaB ligand (RANKL)-induced association between receptor activator of NF-kappaB (RANK) and tumor necrosis factor receptor associated factor 6 (TRAF6), immunofluorescence staining and immunoprecipitation experiments were performed. During our validation model, histomorphometric examination and micro-computed tomography (CT) were conducted to assess the morphology of osteoporosis. RESULTS: Shikonin prevented bone loss by inhibiting osteoclastogenesis in vitro and improving bone loss in ovariectomized mice in vivo. At the molecular level, Western blot analysis indicated that shikonin inhibited the phosphorylation of inhibitor of NF-kappaB (IkappaB), P50, P65, extracellular regulated protein kinases (ERK), c-Jun N-terminal kinase (JNK), and P38. Interaction of TRAF6 and RANK was prevented, and downstream MAPK and NF-kappaB signaling pathways were downregulated. CONCLUSION: Osteoclastic bone resorption was reduced in the presence of shikonin in vitro and in vivo. Shikonin is a promising candidate for treatment of postmenopausal osteoporosis. CI - Copyright (c) 2020 The Author(s). Published by Elsevier Masson SAS.. All rights reserved. FAU - Chen, Kai AU - Chen K AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: 515854039@qq.com. FAU - Yan, Zijun AU - Yan Z AD - Graduate Management Unit, Shanghai Changhai Hospital, Navy Medical University, Shanghai 200433, China. Electronic address: 15821673297@163.com. FAU - Wang, Yiran AU - Wang Y AD - Shanghai Changhai Hospital, Navy Medical University, Shanghai 200433, China. Electronic address: wyrwangyiran@163.com. FAU - Yang, Yilin AU - Yang Y AD - Shanghai Changhai Hospital, Navy Medical University, Shanghai 200433, China. Electronic address: 994237071@qq.com. FAU - Cai, Mengxi AU - Cai M AD - Graduate Management Unit, Shanghai Changhai Hospital, Navy Medical University, Shanghai 200433, China. Electronic address: cicismmu@163.com. FAU - Huang, Chunyou AU - Huang C AD - Graduate Management Unit, Shanghai Changhai Hospital, Navy Medical University, Shanghai 200433, China. Electronic address: hcysmmu@163.com. FAU - Li, Bo AU - Li B AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: smmulibo@163.com. FAU - Yang, Mingyuan AU - Yang M AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: yangmingyuan0330@163.com. FAU - Zhou, Xiaoyi AU - Zhou X AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: 13818909826@163.com. FAU - Wei, Xianzhao AU - Wei X AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: weixianzhao@126.com. FAU - Yang, Changwei AU - Yang C AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: changwei_y@qq.com. FAU - Chen, Ziqiang AU - Chen Z AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: ziqiang_chen81@126.com. FAU - Zhai, Xiao AU - Zhai X AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: zhaixiao@smmu.edu.cn. FAU - Li, Ming AU - Li M AD - Department of Orthopedics, Changhai Hospital of Navy Medical University, Shanghai 200433, China. Electronic address: limingchspine@126.com. LA - eng PT - Journal Article DEP - 20200406 PL - France TA - Biomed Pharmacother JT - Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie JID - 8213295 RN - 0 (Biomarkers) RN - 0 (NF-kappa B) RN - 0 (Naphthoquinones) RN - 0 (RANK Ligand) RN - 0 (Receptor Activator of Nuclear Factor-kappa B) RN - 0 (TNF Receptor-Associated Factor 6) RN - 3IK6592UBW (shikonin) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinases) SB - IM MH - Animals MH - Biomarkers MH - Bone Resorption/drug therapy/*etiology/*metabolism/pathology MH - Cell Differentiation MH - Cell Survival/drug effects MH - Disease Models, Animal MH - Disease Susceptibility MH - Female MH - Fluorescent Antibody Technique MH - Gene Expression Regulation/drug effects MH - Humans MH - Immunohistochemistry MH - Mice MH - Mitogen-Activated Protein Kinases/metabolism MH - Models, Biological MH - NF-kappa B/metabolism MH - Naphthoquinones/chemistry/*pharmacology MH - Osteoclasts/drug effects/metabolism MH - Osteogenesis/*drug effects MH - Osteoporosis, Postmenopausal MH - Ovariectomy/adverse effects MH - Protein Binding MH - RANK Ligand/*metabolism MH - Receptor Activator of Nuclear Factor-kappa B/genetics/metabolism MH - Signal Transduction/*drug effects MH - TNF Receptor-Associated Factor 6/*metabolism MH - X-Ray Microtomography OTO - NOTNLM OT - Osteoclast OT - Osteoclastogensis OT - Osteoporosis OT - Shikonin OT - TRAF6 COIS- Declaration of Competing Interest The authors declare that there are no conflicts of interest. EDAT- 2020/04/10 06:00 MHDA- 2021/01/27 06:00 CRDT- 2020/04/10 06:00 PHST- 2019/11/17 00:00 [received] PHST- 2020/02/27 00:00 [revised] PHST- 2020/03/03 00:00 [accepted] PHST- 2020/04/10 06:00 [pubmed] PHST- 2021/01/27 06:00 [medline] PHST- 2020/04/10 06:00 [entrez] AID - S0753-3322(20)30258-4 [pii] AID - 10.1016/j.biopha.2020.110067 [doi] PST - ppublish SO - Biomed Pharmacother. 2020 Jun;126:110067. doi: 10.1016/j.biopha.2020.110067. Epub 2020 Apr 6.