Lipopolysaccharide (LPS) induces sclerostin secretion by extracellular vesicle via TLR4/miR-92a-3p/PTEN/NF-κB signalling pathway in murine macrophage

被引:0
作者
Kwok, Carsten Tsun-Ka [1 ]
Wong, Chun-Chak [2 ]
Li, Jing-Jing [3 ,4 ]
Kwan, Yiu-Wa [5 ]
Leung, George Pak-Heng [6 ]
Tsoi, Bun [1 ,7 ]
Chow, Franklin Wang-Ngai [8 ]
Seto, Sai-Wang [1 ,7 ,9 ,10 ]
机构
[1] Hong Kong Polytech Univ, Dept Food Sci & Nutr, Hong Kong, Peoples R China
[2] Hong Kong Polytech Univ, Dept Appl Biol & Chem Technol, Hong Kong, Peoples R China
[3] Hong Kong Polytech Univ, Dept Rehabil Sci, Hong Kong, Peoples R China
[4] Hong Kong Polytech Univ, Shenzhen Res Inst, Shenzhen, Peoples R China
[5] Chinese Univ Hong Kong, Sch Biomed Sci, Hong Kong, Peoples R China
[6] Univ Hong Kong, Dept Pharmacol & Pharm, Hong Kong, Peoples R China
[7] Hong Kong Polytech Univ, Res Ctr Chinese Med Innovat, Hong Kong, Peoples R China
[8] Hong Kong Polytech Univ, Dept Hlth Technol & Informat, Hong Kong, Peoples R China
[9] Western Sydney Univ, NICM Hlth Res Inst, Penrith, NSW 2751, Australia
[10] Univ Western Australia, Sch Biomed Sci, Perth, WA, Australia
关键词
Macrophage; Sclerostin; microRNA; 92a-3p; TLR4/NF-kappa B; Inflammation; Extracellular vesicles; TNF-ALPHA; EXPRESSION; MICE; INFLAMMATION; OSTEOCLAST;
D O I
10.1007/s00011-024-01987-1
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
BackgroundSclerostin (SOST) is traditionally regarded as an osteocyte-derived secreted glycoprotein that regulates bone mineralization. Recent studies reported that SOST is also released from non-skeletal sources, especially during inflammation. However, the cellular source and regulatory mechanisms governing SOST generation in inflammation remain unclear. This study investigated whether macrophages produce SOST in response to inflammatory stimuli and determined associated regulatory pathways.MethodsThe effect of lipopolysaccharide (LPS)-induced inflammation in SOST generation and its underlying regulatory mechanism was examined on mouse macrophage RAW 264.7 by western blot and immunofluorescent staining. Transfection with miR-92a-3p mimic and inhibitor were used to validate its role in SOST production. The role of NF-kappa B and TLR4 were studied using pharmacological inhibitors BAY 11-7085 and TAK242, respectively. The involvement of NF-kappa B and TLR4 in LPS-induced SOST production was further validated through nuclear NF-kappa B p65 immunoprecipitation and TLR4 small interfering RNA (siRNA) experiments, respectively.GW4869 and manumycin A (extracellular vesicles (EV) biogenesis inhibitors) were used to examine the associated of SOST and EV. Finally, SOST expression and characteristics of the isolated EV were assessed by Western blot and nanoparticle tracking analysis (NTA).ResultsLPS significantly induced SOST protein expression and secretion in RAW 264.7. MiR-92a-3p was upregulated by LPS stimulation in macrophages. Transfection of miR-92a-3p mimic increased SOST generation in RAW 264.7. Inhibition of TLR4 and NF-kappa B signalling pathways using pharmacological inhibitors significantly suppressed LPS-induced SOST in RAW 264.7. Similarly, TLR4 siRNA effectively suppressed LPS-induced SOST level. However, the LPS-induced upregulation of miR-92a-3p was only regulated by TLR4, but not by NF-kappa B. NF-kappa B was found to directly bind to the mouse sost promoter, thereby activating sost transcription. Additionally, SOST secretion was found predominantly associated with EV from LPS-stimulated cells, and inhibition of EV biogenesis suppressed SOST production in RAW 264.7 cells.ConclusionsIn conclusion, our study showed, for the first time, that LPS induced SOST generation and secretion via TLR4/miR-92a-3p/PTEN/NF-kappa B singling pathway in murine macrophage RAW 264.7 cells. Moreover, we showed that SOST is secreted from the RAW 264.7 cells in the form of extracellular vesicle. This study identified macrophage as a novel source of SOST, highlighting its potential role in inflammatory diseases.
引用
收藏
页数:18
相关论文
共 53 条
  • [1] Almansouri Abdulrahman Y, 2016, J Bone Metab, V23, P16, DOI 10.11005/jbm.2016.23.1.16
  • [2] TNF-α Upregulates Sclerostin Expression in Obese Mice Fed a High-Fat Diet
    Baek, Kyunghwa
    Hwang, Hyo Rin
    Park, Hyun-Jung
    Kwon, Arang
    Qadir, Abdul S.
    Ko, Seong-Hee
    Woo, Kyung Mi
    Ryoo, Hyun-Mo
    Kim, Gwan-Shik
    Baek, Jeong-Hwa
    [J]. JOURNAL OF CELLULAR PHYSIOLOGY, 2014, 229 (05) : 640 - 650
  • [3] MicroRNA-138 Aggravates Inflammatory Responses of Macrophages by Targeting SIRT1 and Regulating the NF-κB and AKT Pathways
    Bai, Xiao-Zhi
    Zhang, Ju-Lei
    Liu, Yang
    Zhang, Wei
    Li, Xiao-Qiang
    Wang, Ke-Jia
    Cao, Meng-Yuan
    Zhang, Jia-Ning
    Han, Fu
    Shi, Ji-Hong
    Hu, Da-Hai
    [J]. CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, 2018, 49 (02) : 489 - 500
  • [4] Extracellular MicroRNA-92a Mediates Endothelial Cell-Macrophage Communication
    Chang, Ya-Ju
    Li, Yi-Shuan
    Wu, Chia-Ching
    Wang, Kuei-Chun
    Huang, Tzu-Chieh
    Chen, Zhen
    Chien, Shu
    [J]. ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2019, 39 (12) : 2492 - 2504
  • [5] The functions of tumor suppressor PTEN in innate and adaptive immunity
    Chen, Lang
    Guo, Deyin
    [J]. CELLULAR & MOLECULAR IMMUNOLOGY, 2017, 14 (07) : 581 - 589
  • [6] Anti-inflammatory effects of miR-150 are associated with the downregulation of STAT1 in macrophages following lipopolysaccharide treatment
    Chen, Song
    Zhu, Haijun
    Sun, Jie
    Zhu, Lili
    Qin, Long
    Wan, Jian
    [J]. EXPERIMENTAL AND THERAPEUTIC MEDICINE, 2021, 22 (04)
  • [7] Sclerostin inhibition reverses systemic, periarticular and local bone loss in arthritis
    Chen, Xiao-Xiang
    Baum, Wolfgang
    Dwyer, Denise
    Stock, Michael
    Schwabe, Kay
    Ke, Hua-Zhu
    Stolina, Marina
    Schett, Georg
    Bozec, Aline
    [J]. ANNALS OF THE RHEUMATIC DISEASES, 2013, 72 (10) : 1732 - 1736
  • [8] Secretion of an Argonaute protein by a parasitic nematode and the evolution of its siRNA guides
    Chow, Franklin Wang-Ngai
    Koutsovoulos, Georgios
    Ovando-Vazquez, Cesare
    Neophytou, Kyriaki
    Bermudez-Barrientos, Jose R.
    Laetsch, Dominik R.
    Robertson, Elaine
    Kumar, Sujai
    Claycomb, Julie M.
    Blaxter, Mark
    Abreu-Goodger, Cei
    Buck, Amy H.
    [J]. NUCLEIC ACIDS RESEARCH, 2019, 47 (07) : 3594 - 3606
  • [9] Sclerostin Protects Against Vascular Calcification Development in Mice
    De Mare, Annelies
    Opdebeeck, Britt
    Neven, Ellen
    D'Haese, Patrick C.
    Verhulst, Anja
    [J]. JOURNAL OF BONE AND MINERAL RESEARCH, 2022, 37 (04) : 687 - 699
  • [10] The Role of Sclerostin in Bone and Ectopic Calcification
    De Mare, Annelies
    D'Haese, Patrick C.
    Verhulst, Anja
    [J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2020, 21 (09)