Neobavaisoflavone ameliorates LPS-induced RAW264.7 cell inflammations by suppressing the activation of NF-κB and MAPKs signaling pathways

Document Type : Original Article

Authors

1 State Key Laboratory of Component-based Chinese Medicine, Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin, China

2 Tianjin Key Laboratory of Traditional Chinese medicine Pharmacology, Tianjin University of Traditional Chinese Medicine, Tianjin, China

3 China Resources Sanjiu Medical & Pharmaceutical Co., Ltd, Shenzhen, China

Abstract

Objective(s): Neobavaisoflavone (NBIF) is an isoflavone isolated from Psoralea corylifolia L. It can effectively regulate the redox state as a natural anti-oxidant and show some anti-inflammatory activity. However, its molecular mechanism is poorly studied. In this study, RAW264.7 cells were treated with lipopolysaccharide (LPS) to investigate the anti-inflammatory activity and potential NBIF mechanism.
Materials and Methods: RAW264.7 cells were treated with LPS (62.5 ng/ml) and exposed to different concentrations of NBIF (0.01, 0.1, and 1 μM) for 24 hr. Inflammatory cytokines of RAW264.7 cells were measured by the Griess method, ELISA, and western blot. Phagocytosis of RAW264.7 macrophages was measured by FITC-dextran uptake assay. The phosphorylation protein expression levels of MAPKs (JNK, p38, and ERK), NF-κB p65, IκBα, and IκB kinase were analyzed by western blot. The results were analyzed using one-way ANOVA with Tukey’s multiple comparison test.
Results: NBIF significantly inhibited NO and ROS production by down-regulation of iNOS and COX-2 protein expression. Additionally, the amount of release and protein levels of inflammation cytokines IL-6, IL-1β, and TNF-α were significantly decreased by NBIF. Moreover, FITC–dextran uptake assay by flow cytometry presented that NBIF significantly enhanced the phagocytic capacity of RAW264.7. Mechanistically, NBIF significantly down-regulated MAPK activation and inhibited the nuclear translocation of NF-κB p65. 
Conclusion: The present study demonstrates that NBIF inhibited inflammation and enhanced the phagocytic capacity of RAW264.7 cell-related MAPKs and NF-κB signaling pathways induced by LPS. These findings suggest that NBIF may have clinical utility as an anti-inflammatory agent.

Keywords

References

1. Zhong J X, Shi G X . Editorial: Regulation of inflammation in chronic disease. Front Immunol 2019; 12:737-739.
2.    Soehnlein O, Libby P. Targeting inflammation in atherosclerosis - from experimental insights to the clinic. Nat Rev Drug Discov 2021; 20:589-610. 
3.    S Watanabe, M Alexander, AV Misharin, Budinger GRS. The role of macrophages in the resolution of inflammation. J Clin Invest 2019; 129:2619-2628.
4.    Hwang JH, Kim KJ, Ryu SJ, Lee BY. Caffeine prevents LPS-induced inflammatory responses in RAW264.7 cells and zebrafish. Chem Biol Interact 2016; 248:1-7.
5.    Feehan KT, Gilroy DW. Is resolution the end of inflammation? Trends Mol Med 2019; 25:198-214.
6.    Fang Y W, Yang L, He J W. Plantanone C attenuates LPS-stimulated inflammation by inhibiting NF-κB/iNOS/COX-2/MAPKs/Akt pathways in RAW 264.7 macrophages. Biomed Pharmacother 2021; 143:112104-112112.
7.    Kourtzelis I, Hajishengallis G, Chavakis T. Phagocytosis of apoptotic cells in resolution of inflammation. Front Immunol 2020; 11:553-561.
8.    Cao Y, Chen J, Ren G, Zhang Y, Tan X, Yang L. Punicalagin prevents inflammation in LPS-induced RAW264.7 macrophages by inhibiting FoxO3a/autophagy signaling pathway. Nutrients 2019; 11:2794-2808. 
9.    Tang B, Li X, Ren Y, Wang J, Xu D, Hang Y, et al. MicroRNA-29a regulates lipopolysaccharide (LPS)-induced inflammatory responses in murine macrophages through the Akt1/ NF-κB pathway. Exp Cell Res 2017; 360:74-80.
10.    Yeung YT, Aziz F, Guerrero-Castilla A, Arguelles S. Signaling pathways in inflammation and anti-inflammatory therapies. Curr Pharm Des 2018; 24:1449-1484.
11.    Shou J, Kong X, Wang X, Tang Y, Wang C, Wang M, et al. Tizoxanide inhibits inflammation in LPS-activated RAW264.7 macrophages via the suppression of NF-κB and MAPK activation. Inflammation 2019; 42:1336-1349. 
12.    Ullah H, Kwon T H, Park S J, Kim SD, Rhee MH. Isoleucilactucin ameliorates coal fly ash-induced inflammation through the NF-κB and MAPK pathways in MH-S cells. Int J Mol Sci 2021; 22:9506-9518.
13.    Alam F, Khan GN, Asad MHHB. Psoralea corylifolia L: Ethnobotanical, biological, and chemical aspects: A review. Phytother Res 2018; 32:597-615. 
14.    Gao Q, Xu Z, Zhao G, et al. Simultaneous quantification of 5 main components of Psoralea corylifolia L. in rats’ plasma by utilizing ultra high pressure liquid chromateap-hy tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2016; 1011:128-135.
15.    Szliszka E, Skaba D, Czuba ZP, Wang H, Weng Z, Pei K, et al. Inhibition of inflammatory mediators by neobavaisoflavone in activated RAW264.7 macrophages. Molecules 2011; 16:3701-3712. 
16.    Szliszka E, Czuba ZP, Sedek L, Paradysz A, Crol W.  Enhanced TRAIL-mediated apopt-osis in prostate cancer cells by the bioactive compounds neobavaisoflavone and psoralidin isolated from Psoralea corylifolia. Pharmacol Rep 2011; 63:139-148. 
17.    Onuora S. Inflammation: potassium channel regulates osteoclastogenesis. Nat Rev Rheumatol 2018; 14:64-65. 
18.    Don M J, Lin L C, Chiou W F. Neobavaisoflavone stimulates osteogenesis via p38-mediated up-regulation of transcription factors and osteoid genes expression in MC3T3-E1 cells. Phytomedicine 2012; 19:551-561. 
19.    Chen H, Fang C, Zhi X, Song S, Gu Y, Chen X, et al. Neobavaisoflavone inhibits osteoclastogenesis through blocking RANKL signalling-mediated TRAF6 and c-Src recruitment and NF-κB, MAPK and Akt pathways. J Cell Mol Med 2020; 24:9067-9084. 
20.    Vijayan V, Pradhan P, Braud L, Fuchs HR, Gueler F, Motterlini R, et al. Human and murine macrophages exhibit differential metabolic responses to lipopolysaccharide – a divergent role for glycolysis. Redox Biol 2019; 22:101147-101156.
21.    Yang Y, Wei Z, Teichmann At, Wieland FH, Wang A, Lei X, et al. Development of a novel nitric oxide (NO) production inhibitor with potential therapeutic effect on chronic inflammation. Eur J Med Chem 2020; 193:112216-112230.
22.    Minhas R, Bansal Y, Bansal G. Inducible nitric oxide synthase inhibitors: A comprehensive update. Med Res Rev 2019; 40:823-855.
23.    Fan G, Jiang X, Wu X, Fordjour PA, Miao L, Zhang H, et al. Anti-inflammatory activity of tanshinone IIA in LPS-stimulated RAW264.7 macrophages via miRNAs and TLR4-NF-κB pathway. Inflammation 2016; 39:375-384.
24.    Rezaei M, Ghafouri H, Aghamaali MR, Shourian M. Thiazolidinedione derivative suppresses LPS- induced COX-2 expression and NO production in RAW 264.7 macro-phages. Iran J Pharm Res 2019; 18:1371-1379.
25.    Ko EY, Heo SJ, Cho SH, Lee WW, Kim SY, Yang HW, et al. 3Bromo5(ethoxymethyl)1,2benzenediol inhibits LPS- induced pro-inflammatory responses by preventing ROS production and downregulating NF-κB in vitro and in a zebrafish model. Int Immunopharmacol 2019; 67:98-105.
26.    Son ES, Park JW, Kim SH, Park HR, Han W, Kwon OC, et al. Antiinflammatory activity of 3,5,6,7,3’,4’hexamethox -yflavone via repression of the NFκB and MAPK signaling pathways in LPS stimulated RAW264.7 cells. Mol Med Rep 2020; 22:1985-1993.
27.    Yu H, Lin L, Zhang Z, Zhang H, Hu H. Targeting NF-κB pathway for the therapy of diseases: mechanism and clinical study. Signal Transduct Target Ther 2020; 5:209-232.
28.    Dorrington M G, Fraser I. NF-κB signaling in macrophages: dynamics, crosstalk, and signal integration. Front Immunol 2019; 10:705-717.
29.    Behl T, Rana T, Alotaibi GH, Shamsuzzaman M, Naqvi M, Sehgal A, et al. Polyphenols inhibiting MAPK signalling pathway mediated oxidative stress and inflammation in depression. Biomed Pharmacother 2022; 146:112545.
30.    Pang M, Yuan Y, Wang D, Li T, Wang D, Shi X, et al. Recombinant CC16 protein inhibits the production of pro-inflammatory cytokines via NF-κB and p38 MAPK pathways in LPS-activated RAW264.7 macrophages. Acta Biochim Biophys Sin (Shanghai) 2017; 49:435-443.
31.    Jung T W, Chung Y H, Kim H C, El-Aty AMA, Jeong JH. LECT2 promotes inflammation and insulin resistance in adipocytes via P38 pathways. J Mol Endocrinol 2018; 61:37-45.
32.    Ouyang S, Yao YH,  Zhang ZM, Liu JS, Xiang H. Curcumin inhibits hypoxia inducible factor-1α-induced inflammation and apoptosis in macrophages through an ERK dependent pathway. Eur Rev Med Pharmacol Sci 2019; 23:1816-1825. 
33.    Guo M, Härtlova A, Gierliński M, Prescott A, Castellvi J, Losa JH, et al. Triggering MSR1 promotes JNK-mediated inflamemation in IL-4-activated macrophages. EMBO J 2019; 38:100299-100314. 
Iranian Journal of Basic Medical Sciences
Volume 25, Issue 8
August 2022
Pages 1021-1027

Files

Share

How to cite

Statistics