Abstract
Nuciferine, isolated from Nelumbo nucifera (commonly known as lotus) leaves, has been shown to have beneficial effects, including antioxidant, anti-obesity, anti-diabetic, and anti-inflammatory properties. However, little is known about the mechanism of nuciferine action on the inflammatory response. This study aimed to investigate the anti-inflammatory effects of nuciferine and its underlying molecular mechanisms in lipopolysaccharide (LPS)-stimulated murine macrophages. In this study, nuciferine reduced LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production and mRNA expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Nuciferine also decreased the production of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. Furthermore, nuciferine inhibited the LPS-mediated transcriptional activity of nuclear factor (NF)-κB and activator protein (AP)-1, and the nuclear translocation of NF-κB p65 and activating transcription factor 2 (ATF2), an AP-1 subunit. Nuciferine also decreased the phosphorylation of IκB kinase (IKK), inhibitor of NF-κB (IκB), NF-κB, mitogen-activated protein kinase 3 (MKK3), MKK6, p38 mitogen-activated protein kinase (MAPK), and ATF2. Overall, our findings suggest that nuciferine may exert anti-inflammatory effects in LPS-induced macrophages by inhibiting the NF-κB and p38 MAPK/ATF2 signaling pathways.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are available from the corresponding author upon reasonable request.
Abbreviations
- VC:
-
Vehicle control
- LPS:
-
Lipopolysaccharide
- NO:
-
Nitric oxide
- PGE2 :
-
Prostaglandin E2
- iNOS:
-
Inducible nitric oxide synthase
- COX-2:
-
Cyclooxygenase-2
- IL-1β:
-
Interleukin-1β
- IL-6:
-
Interleukin 6
- TNF-α:
-
Tumor necrosis factor-α
- IKK:
-
IκB kinas
- IκB:
-
Inhibitor of NF-κB
- NF-κB:
-
Nuclear factor-κB
- MAP2K (also known as MKK):
-
Mitogen-activated protein kinase kinase
- MAPK:
-
Mitogen-activated protein kinase
- ATF2:
-
Activating transcription factor 2
- AP-1:
-
Activator protein 1
- JNK:
-
C-Jun N-terminal kinase
References
Ahn JH, Kim ES, Lee C, Kim S, Cho S-H, Hwang BY et al (2013) Chemical constituents from Nelumbo nucifera leaves and their anti-obesity effects. Bioorg Med Chem Lett 23(12):3604–3608
Bhat NR, Feinstein DL, Shen Q, Bhat AN (2002) p38 MAPK-mediated transcriptional activation of inducible nitric-oxide synthase in glial cells. Roles of nuclear factors, nuclear factor kappa B, cAMP response element-binding protein, CCAAT/enhancer-binding protein-beta, and activating transcription factor-2. J Biol Chem 277(33):29584–29592. https://doi.org/10.1074/jbc.M204994200 (Epub 2002/06/06)
Chen L, Deng H, Cui H, Fang J, Zuo Z, Deng J et al (2018a) Inflammatory responses and inflammation-associated diseases in organs. Oncotarget 9(6):7204–7218. https://doi.org/10.18632/oncotarget.23208 (Epub 2018/02/23)
Chen X, Zheng X, Zhang M, Yin H, Jiang K, Wu H et al (2018b) Nuciferine alleviates LPS-induced mastitis in mice via suppressing the TLR4-NF-kappaB signaling pathway. Inflamm Res 67(11–12):903–911. https://doi.org/10.1007/s00011-018-1183-2 (Epub 2018/08/27)
Chun KS, Kim SH, Song YS, Surh YJ (2004) Celecoxib inhibits phorbol ester-induced expression of COX-2 and activation of AP-1 and p38 MAP kinase in mouse skin. Carcinogenesis 25(5):713–722. https://doi.org/10.1093/carcin/bgh076 (Epub 20040116)
Chun JN, Park S, Lee S, Kim JK, Park EJ, Kang M et al (2018) Schisandrol B and schisandrin B inhibit TGFbeta1-mediated NF-kappaB activation via a Smad-independent mechanism. Oncotarget 9(3):3121–3130. https://doi.org/10.18632/oncotarget.23213 (Epub 2018/02/10)
Fang P, Li X, Dai J, Cole L, Camacho JA, Zhang Y et al (2018) Immune cell subset differentiation and tissue inflammation. J Hematol Oncol 11(1):97. https://doi.org/10.1186/s13045-018-0637-x (Epub 2018/08/02)
Gazon H, Barbeau B, Mesnard JM, Peloponese JM Jr (2017) Hijacking of the AP-1 signaling pathway during development of ATL. Front Microbiol 8:2686. https://doi.org/10.3389/fmicb.2017.02686 (Epub 20180115)
Gozdecka M, Breitwieser W (2012) The roles of ATF2 (activating transcription factor 2) in tumorigenesis. Biochem Soc Trans 40(1):230–234. https://doi.org/10.1042/BST20110630 (Epub 2012/01/21)
Guo F, Yang X, Li X, Feng R, Guan C, Wang Y et al (2013) Nuciferine prevents hepatic steatosis and injury induced by a high-fat diet in hamsters. PLoS ONE 8(5):e63770. https://doi.org/10.1371/journal.pone.0063770 (Epub 2013/05/22)
Gyorfy Z, Duda E, Vizler C (2013) Interactions between LPS moieties and macrophage pattern recognition receptors. Vet Immunol Immunopathol 152(1–2):28–36. https://doi.org/10.1016/j.vetimm.2012.09.020 (Epub 2012/10/23)
Han C, Sun T, Liu Y, Fan G, Zhang W, Liu C (2020) Protective effect of Polygonatum sibiricum polysaccharides on gentamicin-induced acute kidney injury in rats via inhibiting p38 MAPK/ATF2 pathway. Int J Biol Macromol 151:595–601. https://doi.org/10.1016/j.ijbiomac.2020.02.049 (Epub 2020/02/15)
Heo J (2002) Donguibogam. Beob-in Munhwasa, Seoul, Korea, pp 1883–1884
Heumann D, Roger T (2002) Initial responses to endotoxins and Gram-negative bacteria. Clin Chim Acta 323(1–2):59–72. https://doi.org/10.1016/s0009-8981(02)00180-8 (Epub 2002/07/24)
Hollenbach E, Neumann M, Vieth M, Roessner A, Malfertheiner P, Naumann M (2004) Inhibition of p38 MAP kinase- and RICK/NF-kappaB-signaling suppresses inflammatory bowel disease. FASEB J 18(13):1550–1552. https://doi.org/10.1096/fj.04-1642fje (Epub 20040802)
Kim DH, Chung JH, Yoon JS, Ha YM, Bae S, Lee EK et al (2013) Ginsenoside Rd inhibits the expressions of iNOS and COX-2 by suppressing NF-kappaB in LPS-stimulated RAW264.7 cells and mouse liver. J Ginseng Res 37(1):54–63. https://doi.org/10.5142/jgr.2013.37.54
Kim SM, Park EJ, Kim JY, Choi J, Lee HJ (2020) Anti-inflammatory effects of fermented lotus root and linoleic acid in lipopolysaccharide-induced RAW 264.7 Cells. Life (basel). https://doi.org/10.3390/life10110293 (Epub 20201119)
Kumar S, Boehm J, Lee JC (2003) p38 MAP kinases: key signalling molecules as therapeutic targets for inflammatory diseases. Nat Rev Drug Discov 2(9):717–726. https://doi.org/10.1038/nrd1177
Li S, Wang Y, Matsumura K, Ballou LR, Morham SG, Blatteis CM (1999) The febrile response to lipopolysaccharide is blocked in cyclooxygenase-2(-/-), but not in cyclooxygenase-1(-/-) mice. Brain Res 825(1–2):86–94. https://doi.org/10.1016/s0006-8993(99)01225-1
Li Z, Chen Y, An T, Liu P, Zhu J, Yang H et al (2019) Nuciferine inhibits the progression of glioblastoma by suppressing the SOX2-AKT/STAT3-Slug signaling pathway. J Exp Clin Cancer Res 38(1):139. https://doi.org/10.1186/s13046-019-1134-y (Epub 2019/03/30)
Li D, Liu B, Fan Y, Liu M, Han B, Meng Y et al (2021) Nuciferine protects against folic acid-induced acute kidney injury by inhibiting ferroptosis. Br J Pharmacol 178(5):1182–1199. https://doi.org/10.1111/bph.15364 (Epub 2021/01/16)
Lim JS, Oh J, Yun HS, Lee JS, Hahn D, Kim JS (2022) Anti-neuroinflammatory activity of 6,7-dihydroxy-2,4-dimethoxy phenanthrene isolated from Dioscorea batatas Decne partly through suppressing the p38 MAPK/NF-kappaB pathway in BV2 microglial cells. J Ethnopharmacol 282:114633. https://doi.org/10.1016/j.jep.2021.114633 (Epub 2021/09/15)
Liu S, Feng G, Wang GL, Liu GJ (2008) p38MAPK inhibition attenuates LPS-induced acute lung injury involvement of NF-kappaB pathway. Eur J Pharmacol 584(1):159–165. https://doi.org/10.1016/j.ejphar.2008.02.009 (Epub 2008/03/11)
Makarov SS (2000) NF-kappaB as a therapeutic target in chronic inflammation: recent advances. Mol Med Today 6(11):441–448. https://doi.org/10.1016/s1357-4310(00)01814-1
Malemud CJ (2004) Protein kinases in chondrocyte signaling and osteoarthritis. Clin Orthop Relat Res. https://doi.org/10.1097/01.blo.0000143802.41885.50
Ning Q, Wang Y, Zhang Y, Shen G, Xie Z, Pang J (2019) Nuciferine prevents hepatic steatosis by regulating lipid metabolism in diabetic rat model. Open Life Sci. 14:699–706. https://doi.org/10.1515/biol-2019-0079 (Epub 2019/12/31)
Park E-J, You-Suk L, Kim SM, Ah Jin J, Jeong-Hyun Y, Lee S-H et al (2020) Immune-enhancing effects of red Platycodon grandiflorus root extract via p38 MAPK-mediated NF-κB activation. Appl Sci 10(16):5457. https://doi.org/10.3390/app10165457
Paudel KR, Panth N (2015) Phytochemical profile and biological activity of Nelumbo nucifera. Evid-Based Complement Altern Med 2015:789124. https://doi.org/10.1155/2015/789124
Pham TH, Kim MS, Le MQ, Song YS, Bak Y, Ryu HW et al (2017) Fargesin exerts anti-inflammatory effects in THP-1 monocytes by suppressing PKC-dependent AP-1 and NF-kB signaling. Phytomedicine 24:96–103. https://doi.org/10.1016/j.phymed.2016.11.014 (Epub 20161121)
Saha RN, Jana M, Pahan K (2007) MAPK p38 regulates transcriptional activity of NF-kappaB in primary human astrocytes via acetylation of p65. J Immunol 179(10):7101–7109. https://doi.org/10.4049/jimmunol.179.10.7101
Scheidereit C (2006) IkappaB kinase complexes: gateways to NF-kappaB activation and transcription. Oncogene 25(51):6685–6705. https://doi.org/10.1038/sj.onc.1209934
Slomiany BL, Slomiany A (2019) Syk: a new target for attenuation of Helicobacter pylori-induced gastric mucosal inflammatory responses. Inflammopharmacology 27(2):203–211. https://doi.org/10.1007/s10787-019-00577-6 (Epub 20190228)
Wang L, Tu YC, Lian TW, Hung JT, Yen JH, Wu MJ (2006) Distinctive antioxidant and antiinflammatory effects of flavonols. J Agric Food Chem 54(26):9798–9804. https://doi.org/10.1021/jf0620719
Wang MX, Zhao XJ, Chen TY, Liu YL, Jiao RQ, Zhang JH et al (2016) Nuciferine alleviates renal injury by inhibiting inflammatory responses in fructose-fed rats. J Agric Food Chem 64(42):7899–7910. https://doi.org/10.1021/acs.jafc.6b03031 (Epub 2016/10/11)
Watkins LR, Maier SF, Goehler LE (1995) Immune activation: the role of pro-inflammatory cytokines in inflammation, illness responses and pathological pain states. Pain 63(3):289–302. https://doi.org/10.1016/0304-3959(95)00186-7
Watson G, Ronai ZA, Lau E (2017) ATF2, a paradigm of the multifaceted regulation of transcription factors in biology and disease. Pharmacol Res 119:347–357. https://doi.org/10.1016/j.phrs.2017.02.004 (Epub 2017/02/19)
Wu H, Yang Y, Guo S, Yang J, Jiang K, Zhao G et al (2017) Nuciferine ameliorates inflammatory responses by inhibiting the TLR4-mediated pathway in lipopolysaccharide-induced acute lung injury. Front Pharmacol 8:939. https://doi.org/10.3389/fphar.2017.00939 (Epub 20171221)
Yamamoto Y, Gaynor RB (2004) IkappaB kinases: key regulators of the NF-kappaB pathway. Trends Biochem Sci 29(2):72–79. https://doi.org/10.1016/j.tibs.2003.12.003 (Epub 2004/04/23)
Yi YS, Cho JY, Kim D (2016) Cerbera manghas methanol extract exerts anti-inflammatory activity by targeting c-Jun N-terminal kinase in the AP-1 pathway. J Ethnopharmacol 193:387–396. https://doi.org/10.1016/j.jep.2016.08.033 (Epub 20160822)
Yoo JH, Park EJ, Kim SH, Lee HJ (2020) Gastroprotective Effects of fermented lotus root against ethanol/HCl-induced gastric mucosal acute toxicity in rats. Nutrients. https://doi.org/10.3390/nu12030808
Zhang DD, Zhang JG, Wu X, Liu Y, Gu SY, Zhu GH et al (2015) Nuciferine downregulates Per-Arnt-Sim kinase expression during its alleviation of lipogenesis and inflammation on oleic acid-induced hepatic steatosis in HepG2 cells. Front Pharmacol 6:238. https://doi.org/10.3389/fphar.2015.00238 (Epub 2015/11/06)
Zhang C, Deng J, Liu D, Tuo X, Xiao L, Lai B et al (2018a) Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARalpha/PPARgamma coactivator-1alpha pathway. Br J Pharmacol 175(22):4218–4228. https://doi.org/10.1111/bph.14482 (Epub 2018/08/22)
Zhang C, Deng J, Liu D, Tuo X, Yu Y, Yang H et al (2018b) Nuciferine inhibits proinflammatory cytokines via the PPARs in LPS-induced RAW264.7 cells. Molecules. https://doi.org/10.3390/molecules23102723 (Epub 2018/10/27)
Zhang L, Gao J, Tang P, Chong L, Liu Y, Liu P et al (2018c) Nuciferine inhibits LPS-induced inflammatory response in BV2 cells by activating PPAR-γ. Int Immunopharmacol 63:9–13
Acknowledgements
We thank Ju-Hong Jeon (Seoul National University) for providing the 3×κB-Luc reporter gene plasmids.
Funding
This work was supported by the Gachon University research fund of 2022(GCU-202106640001) and Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through the High Value-Added Food Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (grant number 321024-04-1-HD020).
Author information
Authors and Affiliations
Contributions
Conceptualization: E-JP and H-JL; formal analysis: S-MK and E-JP; funding acquisition: H-JL; investigation: S-MK; methodology: S-MK and E-JP; project administration: E-JP; supervision: E-JP and H-JL; validation: S-MK; visualization: S-MK and E-JP; writing—original draft preparation: S-MK; writing—review and editing: E-JP and H-JL.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kim, SM., Park, EJ. & Lee, HJ. Nuciferine attenuates lipopolysaccharide-stimulated inflammatory responses by inhibiting p38 MAPK/ATF2 signaling pathways. Inflammopharmacol 30, 2373–2383 (2022). https://doi.org/10.1007/s10787-022-01075-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10787-022-01075-y