Advertisement

Fisetin rescues retinal functions by suppressing inflammatory response in a DBA/2J mouse model of glaucoma

  • Linlin Li
  • Jie Qin
  • Tingting Fu
  • Jiaxiang ShenEmail author
Original Research Article

Abstract

Purpose

Glaucoma is a common chronic neurodegenerative disease, which could lead to visual loss. In this study, we aimed to investigate whether fisetin, a natural flavone with anti-inflammatory and antioxidant properties, is able to alleviate glaucoma.

Methods

We employed a DBA/2J mouse model which was treated with or without fisetin. Pattern electroretinogram (P-ERG), visual evoked potentials (VEPs) and intraocular pressure (IOP) were evaluated. Quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression levels of TNF-α, IL-1β and IL-6. Western blotting was performed to assess the activation of nuclear factor kappa-B (NF-κB).

Results

We found that DBA/2J mice treated with fisetin (10-30 mg/kg) showed improved P-ERG and VEP amplitudes and reduced IOP compared to untreated DBA/2J mice. In addition, there were more survived retinal ganglion cells (RGCs) and less activated microglia in fisetin-treated DBA/2J mice than those in untreated mice. Furthermore, secreted protein levels and mRNA levels of TNF-α, IL-1β and IL-6 were significantly repressed by fisetin. The phosphorylated p65 level in the nucleus was dramatically reduced in fisetin-treated mice compared to it in untreated mice. Our results demonstrate that fisetin may exert its function through regulating cytokine productions and inhibiting NF-κB activation in the retina.

Conclusion

In conclusion, fisetin is able to promote the visual functions of DBA/2J mice by inhibiting NF-κB activation.

Keywords

Glaucoma Fisetin Retinal ganglion cells (RGCs) Nuclear factor kappa-B (NF-κB) DBA/2J mouse 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving animal were in accordance with the ethical standards of the institutional and/or national research committee. Animal studies were approved by the People’s Hospital of Rizhao, and all efforts were made to minimize the number of animals used and their suffering.

Statements of human rights

There is no human participation in this study.

Statement on the welfare of animals

All animal experiments were conducted according to the guidelines in our institute and all efforts were made to minimize the nubmer of animals used and their suffering.

Informed consent

Not applicable. There is no human participation in this study.

References

  1. 1.
    Quigley HA, Broman AT (2006) The number of people with glaucoma worldwide in 2010 and 2020. Br J Ophthalmol 90:262–267.  https://doi.org/10.1136/bjo.2005.081224 Google Scholar
  2. 2.
    Nuschke AC, Farrell SR, Levesque JM, Chauhan BC (2015) Assessment of retinal ganglion cell damage in glaucomatous optic neuropathy: axon transport, injury and soma loss. Exp Eye Res 141:111–124.  https://doi.org/10.1016/j.exer.2015.06.006 Google Scholar
  3. 3.
    Howell GR, Macalinao DG, Sousa GL, Walden M, Soto I, Kneeland SC, Barbay JM, King BL, Marchant JK, Hibbs M, Stevens B, Barres BA, Clark AF, Libby RT, John SWM (2011) Molecular clustering identifies complement and endothelin induction as early events in a mouse model of glaucoma. J Clin Investig 121:1429–1444.  https://doi.org/10.1172/Jci44646 Google Scholar
  4. 4.
    Nickells RW, Howell GR, Soto I, John SWM (2012) Under pressure: cellular and molecular responses during glaucoma, a common neurodegeneration with axonopathy. Annu Rev Neurosci 35(35):153–179.  https://doi.org/10.1146/annurev.neuro.051508.135728 Google Scholar
  5. 5.
    Foster PJ, Buhrmann R, Quigley HA, Johnson GJ (2002) The definition and classification of glaucoma in prevalence surveys. Br J Ophthalmol 86:238–242.  https://doi.org/10.1136/bjo.86.2.238 Google Scholar
  6. 6.
    Wax MB, Tezel G, Yang J, Peng G, Patil RV, Agarwal N, Sappington RM, Calkins DJ (2008) Induced autoimmunity to heat shock proteins elicits glaucomatous loss of retinal ganglion cell neurons via activated T-cell-derived fas-ligand. J Neurosci 28:12085–12096.  https://doi.org/10.1523/Jneurosci.3200-08.2008 Google Scholar
  7. 7.
    Tezel G, Thornton IL, Tong MG, Luo C, Yang XJ, Cai J, Powell DW, Soltau JB, Liebmann JM, Ritch R (2012) Immunoproteomic analysis of potential serum biomarker candidates in human glaucoma. Invest Ophthalmol Vis Sci 53:8222–8231.  https://doi.org/10.1167/iovs.12-10076 Google Scholar
  8. 8.
    Tatton W, Chen D, Chalmers-Redman R, Wheeler L, Nixon R, Tatton N (2003) Hypothesis for a common basis for neuroprotection in glaucoma and Alzheimer’s disease: anti-apoptosis by alpha-2-adrenergic receptor activation. Surv Ophthalmol 48:S25–S37.  https://doi.org/10.1016/S0039-6257(03)00005-5 Google Scholar
  9. 9.
    Crish SD, Sappington RM, Inman DM, Horner PJ, Calkins DJ (2010) Distal axonopathy with structural persistence in glaucomatous neurodegeneration. Proc Natl Acad Sci USA 107:5196–5201.  https://doi.org/10.1073/pnas.0913141107 Google Scholar
  10. 10.
    Murphy JA, Clarke DB (2006) Target-derived neurotrophins may influence the survival of adult retinal ganglion cells when local neurotrophic support is disrupted: implications for glaucoma. Med Hypotheses 67:1208–1212.  https://doi.org/10.1016/j.mehy.2006.04.049 Google Scholar
  11. 11.
    Zhang HF, Lin Y, Li JH, Pober JS, Min W (2007) RIP1-mediated AIP1 phosphorylation at a 14-3-3-binding site is critical for tumor necrosis factor-induced ASK1-JNK/p38 activation. J Biol Chem 282:14788–14796Google Scholar
  12. 12.
    Wax MB, Tezel G (2009) Immunoregulation of retinal ganglion cell fate in glaucoma. Exp Eye Res 88:825–830.  https://doi.org/10.1016/j.exer.2009.02.005 Google Scholar
  13. 13.
    Nakazawa T, Nakazawa C, Matsubara A, Noda K, Hisatomi T, She HC, Michaud N, Hafezi-Moghadam A, Miller JW, Benowitz LI (2006) Tumor necrosis factor-alpha mediates oligodendrocyte death and delayed retinal ganglion cell loss in a mouse model of glaucoma. J Neurosci 26:12633–12641.  https://doi.org/10.1523/Jneurosci.2801-06.2006 Google Scholar
  14. 14.
    Lebrun-Julien F, Bertrand MJ, De Backer O, Stellwagen D, Morales CR, Di Polo A, Barker PA (2010) ProNGF induces TNFα-dependent death of retinal ganglion cells through a p75NTR non-cell-autonomous signaling pathway. Proc Natl Acad Sci 107:3817–3822Google Scholar
  15. 15.
    Funayama T, Ishikawa K, Ohtake Y, Tanino T, Kurosaka D, Kimura I, Suzuki K, Ideta H, Nakamoto K, Yasuda N, Fujimaki T, Murakami A, Asaoka R, Hotta Y, Tanihara H, Kanamoto T, Mishima H, Fukuchi T, Abe H, Iwata T, Shimada N, Kudoh J, Shimizu N, Mashima Y (2004) Variants in optineurin gene and their association with tumor necrosis factor-alpha polymorphisms in Japanese patients with glaucoma. Invest Ophthalmol Vis Sci 45:4359–4367.  https://doi.org/10.1167/iovs.03-1403 Google Scholar
  16. 16.
    Pal HC, Athar M, Elmets CA, Afaq F (2015) Fisetin inhibits UVB-induced cutaneous inflammation and activation of PI3 K/AKT/NFκB signaling pathways in SKH-1 hairless mice. Photochem Photobiol 91:225–234Google Scholar
  17. 17.
    Kim SC, Kang SH, Jeong SJ, Kim SH, Ko HS, Kim SH (2012) Inhibition of c-Jun N-terminal kinase and nuclear factor kappa B pathways mediates fisetin-exerted anti-inflammatory activity in lipopolysaccharide-treated RAW264.7 cells. Immunopharmacol Immunotoxicol 34:645–650.  https://doi.org/10.3109/08923973.2011.648270 Google Scholar
  18. 18.
    Chuang JY, Chang PC, Shen YC, Lin CJ, Tsai CF, Chen JH, Yeh WL, Wu LH, Lin HY, Liu YS, Lu DY (2014) Regulatory effects of fisetin on microglial activation. Molecules 19:8820–8839.  https://doi.org/10.3390/molecules19078820 Google Scholar
  19. 19.
    Lee JD, Huh JE, Jeon G, Yang HR, Woo HS, Choi DY, Park DS (2009) Flavonol-rich RVHxR from Rhus verniciflua Stokes and its major compound fisetin inhibits inflammation-related cytokines and angiogenic factor in rheumatoid arthritic fibroblast-like synovial cells and in vivo models. Int Immunopharmacol 9:268–276.  https://doi.org/10.1016/j.intimp.2008.11.005 Google Scholar
  20. 20.
    Goh FY, Upton N, Guan S, Cheng C, Shanmugam MK, Sethi G, Leung BP, Wong WF (2012) Fisetin, a bioactive flavonol, attenuates allergic airway inflammation through negative regulation of NF-κB. Eur J Pharmacol 679:109–116Google Scholar
  21. 21.
    Porciatti V, Saleh M, Nagaraju M (2007) The pattern electroretinogram as a tool to monitor progressive retinal ganglion cell dysfunction in the DBA/2J mouse model of glaucoma. Invest Ophthalmol Vis Sci 48:745–751.  https://doi.org/10.1167/iovs.06-0733 Google Scholar
  22. 22.
    Gustafson E, Silberschmidt A, Esguerra M, Miller R (2013) Recording and manipulation of the pattern electroretinogram in a mouse eyecup preparation. Investig Ophthalmol Vis Sci 54:6132Google Scholar
  23. 23.
    Nadal-Nicolas FM, Jimenez-Lopez M, Salinas-Navarro M, Sobrado-Calvo P, Alburquerque-Bejar JJ, Vidal-Sanz M, Agudo-Barriuso M (2012) Whole number, distribution and co-expression of Brn3 transcription factors in retinal ganglion cells of adult albino and pigmented rats. PLoS ONE 7:e49830.  https://doi.org/10.1371/journal.pone.0049830 Google Scholar
  24. 24.
    Libby RT, Gould DB, Anderson MG, John SWM (2005) Complex genetics of glaucoma susceptibility. Annu Rev Genom Hum Genet 6:15–44.  https://doi.org/10.1146/annurev.genom.6.080604.162209 Google Scholar
  25. 25.
    Anderson MG, Libby RT, Mao M, Cosma IM, Wilson LA, Smith RS, John SWM (2006) Genetic context determines susceptibility to intraocular pressure elevation in a mouse pigmentary glaucoma. BMC Biol 4:20.  https://doi.org/10.1186/1741-7007-4-20 Google Scholar
  26. 26.
    Libby RT, Li Y, Savinova OV, Barter J, Smith RS, Nickells RW, John SWM (2005) Susceptibility to neurodegeneration in a glaucoma is modified by Bax gene dosage. PLoS Genet 1:17–26.  https://doi.org/10.1371/journal.pgen.0010004 Google Scholar
  27. 27.
    Reichstein D, Ren LZ, Filippopoulos T, Mittag T, Danias J (2007) Apoptotic retinal ganglion cell death in the DBA/2 mouse model of glaucoma. Exp Eye Res 84:13–21.  https://doi.org/10.1016/j.exer.2006.08.009 Google Scholar
  28. 28.
    Bosco A, Inman DM, Steele MR, Wu GM, Soto I, Marsh-Armstrong N, Hubbard WC, Calkins DJ, Horner PJ, Vetter ML (2008) Reduced retina microglial activation and improved optic nerve integrity with minocycline treatment in the DBA/2J mouse model of glaucoma. Invest Ophthalmol Vis Sci 49:1437–1446.  https://doi.org/10.1167/iovs.07-1337 Google Scholar
  29. 29.
    Neufeld AH (1999) Microglia in the optic nerve head and the region of parapapillary chorioretinal atrophy in glaucoma. Arch Ophthalmol 117:1050–1056Google Scholar
  30. 30.
    Son JL, Soto I, Oglesby E, Lopez-Roca T, Pease ME, Quigley HA, Marsh-Armstrong N (2010) Glaucomatous optic nerve injury involves early astrocyte reactivity and late oligodendrocyte loss. Glia 58:780–789.  https://doi.org/10.1002/glia.20962 Google Scholar
  31. 31.
    Neufeld AH, Liu B (2003) Glaucomatous optic neuropathy: when glia misbehave. Neurosci 9:485–495Google Scholar
  32. 32.
    Sobrado-Calvo P, Vidal-Sanz M, Villegas-Pérez MP (2007) Rat retinal microglial cells under normal conditions, after optic nerve section, and after optic nerve section and intravitreal injection of trophic factors or macrophage inhibitory factor. J Comp Neurol 501:866–878Google Scholar
  33. 33.
    Karlstetter M, Ebert S, Langmann T (2010) Microglia in the healthy and degenerating retina: insights from novel mouse models. Immunobiology 215:685–691.  https://doi.org/10.1016/j.imbio.2010.05.010 Google Scholar
  34. 34.
    Tezel G, Yang XJ, Yang JJ, Wax MB (2004) Role of tumor necrosis factor receptor-1 in the death of retinal ganglion cells following optic nerve crush injury in mice. Brain Res 996:202–212.  https://doi.org/10.1016/j.brainres.2003.10.029 Google Scholar
  35. 35.
    Al-Gayyar M, Elsherbiny N (2013) Contribution of TNF-α to the development of retinal neurodegenerative disorders. Eur Cytokine Netw 24:27–36Google Scholar
  36. 36.
    Balaiya S, Edwards J, Tillis T, Khetpal V, Chalam KV (2011) Tumor necrosis factor-alpha (TNF-α) levels in aqueous humor of primary open angle glaucoma. Clin Ophthalmol 5:553Google Scholar
  37. 37.
    Tezel G (2008) TNF-α signaling in glaucomatous neurodegeneration. Prog Brain Res 173:409–421Google Scholar
  38. 38.
    Wilson GN, Inman DM, Dengler-Crish CM, Smith MA, Crish SD (2015) Early pro-inflammatory cytokine elevations in the DBA/2J mouse model of glaucoma. J Neuroinflammation 12:176Google Scholar
  39. 39.
    H-l Peng, Huang W-C, S-c Cheng, Liou C-J (2018) Fisetin inhibits the generation of inflammatory mediators in interleukin-1β–induced human lung epithelial cells by suppressing the Nf-κb and Erk1/2 pathways. Int Immunopharmacol 60:202–210Google Scholar
  40. 40.
    Sahu BD, Kumar JM, Sistla R (2016) Fisetin, a dietary flavonoid, ameliorates experimental colitis in mice: relevance of NF-κB signaling. J Nutr Biochem 28:171–182Google Scholar
  41. 41.
    Seo S-H, Jeong G-S (2015) Fisetin inhibits TNF-α-induced inflammatory action and hydrogen peroxide-induced oxidative damage in human keratinocyte HaCaT cells through PI3 K/AKT/Nrf-2-mediated heme oxygenase-1 expression. Int Immunopharmacol 29:246–253Google Scholar
  42. 42.
    Mookherjee S, Banerjee D, Chakraborty S, Banerjee A, Mukhopadhyay I, Sen A, Ray K (2010) Association of IL1A and IL1B loci with primary open angle glaucoma. BMC Med Genet 11:99.  https://doi.org/10.1186/1471-2350-11-99 Google Scholar
  43. 43.
    Chidlow G, Wood JPM, Ebneter A, Casson RJ (2012) Interleukin-6 is an efficacious marker of axonal transport disruption during experimental glaucoma and stimulates neuritogenesis in cultured retinal ganglion cells. Neurobiol Dis 48:568–581.  https://doi.org/10.1016/j.nbd.2012.07.026 Google Scholar
  44. 44.
    Li GR, Luna C, Liton PB, Navarro I, Epstein DL, Gonzalez P (2007) Sustained stress response after oxidative stress in trabecular meshwork cells. Mol Vis 13:2282–2288Google Scholar
  45. 45.
    Lebrun-Julien F, Duplan L, Pernet V, Osswald I, Sapieha P, Bourgeois P, Dickson K, Bowie D, Barker PA, Di Polo A (2009) Excitotoxic death of retinal neurons in vivo occurs via a non-cell-autonomous mechanism. J Neurosci 29:5536–5545.  https://doi.org/10.1523/Jneurosci.0831-09.2009 Google Scholar
  46. 46.
    Chen C, Yao L, Cui J, Liu B (2018) Fisetin protects against intracerebral hemorrhage-induced neuroinflammation in aged mice. Cerebrovasc Dis 45:154–161.  https://doi.org/10.1159/000488117 Google Scholar
  47. 47.
    Sandireddy R, Yerra VG, Komirishetti P, Areti A, Kumar A (2016) Fisetin imparts neuroprotection in experimental diabetic neuropathy by modulating Nrf2 and NF-κB pathways. Cell Mol Neurobiol 36:883–892Google Scholar
  48. 48.
    Feng G, Z-y Jiang, Sun B, Fu J, T-z Li (2016) Fisetin alleviates lipopolysaccharide-induced acute lung injury via TLR4-mediated NF-κB signaling pathway in rats. Inflammation 39:148–157Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Linlin Li
    • 1
  • Jie Qin
    • 2
  • Tingting Fu
    • 2
  • Jiaxiang Shen
    • 1
    Email author
  1. 1.Department of OphthalmologyPeople’s Hospital of RizhaoRizhaoChina
  2. 2.Department of OphthalmologyRizhao Central Hospital of ShandongRizhaoChina

Personalised recommendations