Abstract
Microglia, resident innate immune cells in central nervous system, regulates neuroinflammation and is associated with a variety of neuropathologies. The present study investigated the antineuroinflammatory effects of hispidulin (HPD), a naturally flavone compound, in lipopolysaccharide- (LPS-) stimulated BV2 microglia cells. The expression levels of nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory factors were determined by the Griess method, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). Western blotting was used to measure various transcription factors such as Akt, nuclear factor-kappa B (NF-κB), and signal transducer and activator of transcription 3 (STAT3) activities. Our experimental results demonstrated that HPD increased cell viability and reduced apoptosis in LPS-treated BV2 microglia cells. Moreover, HPD significantly reduced the levels of NO, ROS, inducible nitric oxide synthase (iNOS), cyclooxygenase- (COX-) 2, tumor necrosis factor- (TNF-) α, interleukin- (IL-) 1β, IL-6, and prostaglandin E2 (PGE2) in a dose-dependent manner. Phosphorylation of NF-κB/IκB, Akt, and STAT3 proteins expression by HPD was suppressed in LPS-induced BV2 microglial cells. We concluded that HPD may inhibit neuroinflammatory responses by inhibiting NF-κB pathway activation and ROS formation. These results propose that HPD has potential as anti-inflammatory agents against microglia-mediated neuroinflammatory disorders.
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Abbreviations
- LPS:
-
lipopolysaccharide
- HPD:
-
hispidulin
- ROS:
-
reactive oxygen species
- COX-2:
-
cyclooxygenase-2
- ELISA:
-
enzyme-linked immunosorbent assay
- FBS:
-
fetal bovine serum
- IL-1β:
-
interleukin-1β
- IL6:
-
interleukin-6
- TNF-α:
-
tumor necrosis factor
- iNOS:
-
inducible NO synthase
- NF-κB:
-
nuclear factor-κB
- NO:
-
nitric oxide
- PGE2:
-
prostaglandin E2
- STAT3:
-
signal transducer and activator of transcription 3
- MTT:
-
3-(4, 5-dimethylthiazol- 2-yl)- 2, 5-diphenyl tetrazolium bromide
References
Atif M et al (2016) Pharmacological assessment of hispidulin - a natural bioactive flavone. Acta Pol Pharm 73:565–578
Bachiller S, Jiménez-Ferrer I, Paulus A, Yang Y, Swanberg M, Deierborg T, Boza-Serrano A (2018) Microglia in neurological diseases: a road map to brain-disease dependent-inflammatory response. Front Cell Neurosci 12:488. https://doi.org/10.3389/fncel.2018.00488
Block ML, Zecca L, Hong JS (2007) Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 8:57–69. https://doi.org/10.1038/nrn2038
Burton MD, Sparkman NL, Johnson RW (2011) Inhibition of interleukin-6 trans-signaling in the brain facilitates recovery from lipopolysaccharide-induced sickness behavior. J Neuroinflammation 8:54. https://doi.org/10.1186/1742-2094-8-54
Chen X, Yan X, Guo L (2018) Inhibitory effect of Patrinia on BRL-3A cell apoptosis through the TLR4/PI3K/AKT/GSK3beta and TLR4/P38/JNK signaling pathways. Mol Med Rep 17:5344–5349. https://doi.org/10.3892/mmr.2018.8466
Choi HE, Kwak HJ, Kim SK, Cheon HG (2018) Foenumoside B isolated from Lysimachia foenum-graecum extract suppresses LPS-induced inflammatory response via NF-kappaB/AP-1 inactivation in murine macrophages and in endotoxin-induced shock model. Eur J Pharmacol 832:120–128. https://doi.org/10.1016/j.ejphar.2018.05.022
Cianciulli A et al (2016) PI3k/Akt signalling pathway plays a crucial role in the anti-inflammatory effects of curcumin in LPS-activated microglia. Int Immunopharmacol 36:282–290. https://doi.org/10.1016/j.intimp.2016.05.007
Clavin M et al (2007) Anti-inflammatory activity of flavonoids from Eupatorium arnottianum. J Ethnopharmacol 112:585–589. https://doi.org/10.1016/j.jep.2007.04.007
Consonni A, Morara S, Codazzi F, Grohovaz F, Zacchetti D (2011) Inhibition of lipopolysaccharide-induced microglia activation by calcitonin gene related peptide and adrenomedullin. Mol Cell Neurosci 48:151–160. https://doi.org/10.1016/j.mcn.2011.07.006
Dai Y et al (2011) Disruption of IkappaB kinase (IKK)-mediated RelA serine 536 phosphorylation sensitizes human multiple myeloma cells to histone deacetylase (HDAC) inhibitors. J Biol Chem 286:34036–34050. https://doi.org/10.1074/jbc.M111.284216
Frakes AE et al (2014) Microglia induce motor neuron death via the classical NF-kappaB pathway in amyotrophic lateral sclerosis. Neuron 81:1009–1023. https://doi.org/10.1016/j.neuron.2014.01.013
Gao H, Jiang Q, Han Y, Peng J, Wang C (2015) Hispidulin potentiates the antitumor effect of sunitinib against human renal cell carcinoma in laboratory models. Cell Biochem Biophys 71:757–764. https://doi.org/10.1007/s12013-014-0260-6
Ham HJ et al (2019) Bee venom phospholipase A2 ameliorates amyloidogenesis and neuroinflammation through inhibition of signal transducer and activator of transcription-3 pathway in Tg2576 mice. Transl Neurodegener 8:26. https://doi.org/10.1186/s40035-019-0167-7
Hansen DV, Hanson JE, Sheng M (2018) Microglia in Alzheimer’s disease. J Cell Biol 217:459–472. https://doi.org/10.1083/jcb.201709069
He L et al (2011) Hispidulin, a small flavonoid molecule, suppresses the angiogenesis and growth of human pancreatic cancer by targeting vascular endothelial growth factor receptor 2-mediated PI3K/Akt/mTOR signaling pathway. Cancer Sci 102:219–225. https://doi.org/10.1111/j.1349-7006.2010.01778.x
Hsing CH, Lin MC, Choi PC, Huang WC, Kai JI, Tsai CC, Cheng YL, Hsieh CY, Wang CY, Chang YP, Chen YH, Chen CL, Lin CF (2011) Anesthetic propofol reduces endotoxic inflammation by inhibiting reactive oxygen species-regulated Akt/IKKbeta/NF-kappaB signaling. PLoS One 6:e17598. https://doi.org/10.1371/journal.pone.0017598
Kang CH et al (2012) Caffeine suppresses lipopolysaccharide-stimulated BV2 microglial cells by suppressing Akt-mediated NF-kappaB activation and ERK phosphorylation. Food Chem Toxicol 50:4270–4276. https://doi.org/10.1016/j.fct.2012.08.041
Kelly KA, Michalovicz LT, Miller JV, Castranova V, Miller DB, O'Callaghan JP (2018) Prior exposure to corticosterone markedly enhances and prolongs the neuroinflammatory response to systemic challenge with LPS. PLoS One 13:e0190546. https://doi.org/10.1371/journal.pone.0190546
Kim N et al (2018) Piperlongumine inhibits neuroinflammation via regulating NF-kappaB signaling pathways in lipopolysaccharide-stimulated BV2 microglia cells. J Pharmacol Sci 137:195–201. https://doi.org/10.1016/j.jphs.2018.06.004
Leyva-Lopez N, Gutierrez-Grijalva EP, Ambriz-Perez DL, Heredia JB (2016) Flavonoids as cytokine modulators: a possible therapy for inflammation-related diseases. Int J Mol Sci 17. https://doi.org/10.3390/ijms17060921
Lin YC et al (2010) Hispidulin potently inhibits human glioblastoma multiforme cells through activation of AMP-activated protein kinase (AMPK). J Agric Food Chem 58:9511–9517. https://doi.org/10.1021/jf1019533
Montoya T et al (2018) Peracetylated hydroxytyrosol, a new hydroxytyrosol derivate, attenuates LPS-induced inflammatory response in murine peritoneal macrophages via regulation of non-canonical inflammasome, Nrf2/HO1 and JAK/STAT signaling pathways. J Nutr Biochem 57:110–120. https://doi.org/10.1016/j.jnutbio.2018.03.014
Muhammad T, Ikram M, Ullah R, Rehman SU, Kim MO (2019) Hesperetin, a citrus flavonoid, attenuates LPS-induced neuroinflammation, apoptosis and memory impairments by modulating TLR4/NF-kappaB signaling. Nutrients 11. https://doi.org/10.3390/nu11030648
Murakami A, Ohigashi H (2007) Targeting NOX, INOS and COX-2 in inflammatory cells: chemoprevention using food phytochemicals. Int J Cancer 121:2357–2363. https://doi.org/10.1002/ijc.23161
Murase S, McKay RD (2014) Neuronal activity-dependent STAT3 localization to nucleus is dependent on Tyr-705 and Ser-727 phosphorylation in rat hippocampal neurons. Eur J Neurosci 39:557–565. https://doi.org/10.1111/ejn.12412
Nam HY et al (2018) Ibrutinib suppresses LPS-induced neuroinflammatory responses in BV2 microglial cells and wild-type mice. J Neuroinflammation 15:271. https://doi.org/10.1186/s12974-018-1308-0
Norris GT, Kipnis J (2019) Immune cells and CNS physiology: microglia and beyond. J Exp Med 216:60–70. https://doi.org/10.1084/jem.20180199
Pan MH, Lai CS, Ho CT (2010) Anti-inflammatory activity of natural dietary flavonoids. Food Funct 1:15–31. https://doi.org/10.1039/c0fo00103a
Panthi S, Manandhar S, Gautam K (2018) Hydrogen sulfide, nitric oxide, and neurodegenerative disorders. Transl Neurodegener 7:3. https://doi.org/10.1186/s40035-018-0108-x
Park JC, Yoo H, Kim CE, Shim SY, Lee M (2017) Hispidulin-7-O-neohesperidoside from Cirsium japonicum var. ussuriense attenuates the production of inflammatory mediators in LPS-induced raw 264.7 cells and HT-29 cells. Pharmacogn Mag 13:707–711. https://doi.org/10.4103/0973-1296.218116
Patel K, Patel DK (2017) Medicinal importance, pharmacological activities, and analytical aspects of hispidulin: a concise report. J Tradit Complement Med 7:360–366. https://doi.org/10.1016/j.jtcme.2016.11.003
Rahimifard M et al (2017) Targeting the TLR4 signaling pathway by polyphenols: a novel therapeutic strategy for neuroinflammation. Ageing Res Rev 36:11–19. https://doi.org/10.1016/j.arr.2017.02.004
Rau CS et al (2018) Effect of low-fat diet in obese mice lacking toll-like receptors. Nutrients 10. https://doi.org/10.3390/nu10101464
Roy A et al (2016) Potential therapeutic targets for inflammation in toll-like receptor 4 (TLR4)-mediated signaling pathways. Int Immunopharmacol 40:79–89. https://doi.org/10.1016/j.intimp.2016.08.026
Ryan JC, Cross CA, Van Dolah FM (2011) Effects of COX inhibitors on neurodegeneration and survival in mice exposed to the marine neurotoxin domoic acid. Neurosci Lett 487:83–87. https://doi.org/10.1016/j.neulet.2010.10.001
Shih RH, Wang CY, Yang CM (2015) NF-kappaB signaling pathways in neurological inflammation: a mini review. Front Mol Neurosci 8:77. https://doi.org/10.3389/fnmol.2015.00077
Song GJ, Suk K (2017) Pharmacological modulation of functional phenotypes of microglia in neurodegenerative diseases. Front Aging Neurosci 9:139. https://doi.org/10.3389/fnagi.2017.00139
Wang J, Wang J, Wang J, Yang B, Weng Q, He Q (2019) Targeting microglia and macrophages: a potential treatment strategy for multiple sclerosis. Front Pharmacol 10:286. https://doi.org/10.3389/fphar.2019.00286
Weidemann A, Johnson RS (2008) Biology of HIF-1alpha. Cell Death Differ 15:621–627. https://doi.org/10.1038/cdd.2008.12
Xie J et al (2015) Hispidulin prevents hypoxia-induced epithelial-mesenchymal transition in human colon carcinoma cells. Am J Cancer Res 5:1047–1061
Yi JH, Park SW, Kapadia R, Vemuganti R (2007) Role of transcription factors in mediating post-ischemic cerebral inflammation and brain damage. Neurochem Int 50:1014–1027. https://doi.org/10.1016/j.neuint.2007.04.019
Yu DK, Lee B, Kwon M, Yoon N, Shin T, Kim NG, Choi JS, Kim HR (2015) Phlorofucofuroeckol B suppresses inflammatory responses by down-regulating nuclear factor kappaB activation via Akt, ERK, and JNK in LPS-stimulated microglial cells. Int Immunopharmacol 28:1068–1075. https://doi.org/10.1016/j.intimp.2015.08.028
Yu CI et al (2018) Sandensolide induces oxidative stress-mediated apoptosis in oral cancer cells and in zebrafish xenograft model. Mar Drugs 16. https://doi.org/10.3390/md16100387
Zhao D, Kwon SH, Chun YS, Gu MY, Yang HO (2017) Anti-neuroinflammatory effects of fucoxanthin via inhibition of Akt/NF-kappaB and MAPKs/AP-1 pathways and activation of PKA/CREB pathway in lipopolysaccharide-activated BV-2 microglial cells. Neurochem Res 42:667–677. https://doi.org/10.1007/s11064-016-2123-6
Funding
This study was funded by grants from the Tainan Sin-Lau Medical Foundation, the Presbyterian Church in Taiwan (SLH-107-09), the Ministry of Science and Technology of Taiwan (MOST 108-2320-B-037-018), the Chi-Mei Medical Center Liouying Research Grant (CLFHR10735; CLFHR10733), and the Kaohsiung Medical University Research Foundation (KMU-Q108014; KMU-Q108023).
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Yu, CI., Cheng, CI., Kang, YF. et al. Hispidulin Inhibits Neuroinflammation in Lipopolysaccharide-Activated BV2 Microglia and Attenuates the Activation of Akt, NF-κB, and STAT3 Pathway. Neurotox Res 38, 163–174 (2020). https://doi.org/10.1007/s12640-020-00197-x
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DOI: https://doi.org/10.1007/s12640-020-00197-x