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Journal of Neuroinflammation
Published in: Journal of Neuroinflammation 1/2020

Open Access 01-12-2020 | Research

Indole-3-carbinol regulates microglia homeostasis and protects the retina from degeneration

Authors: Amir Saeed Khan, Thomas Langmann

Published in: Journal of Neuroinflammation | Issue 1/2020

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Abstract

Background

Retinal degenerative diseases significantly contribute to visual impairment and blindness. Microglia reactivity is a hallmark of neurodegenerative diseases including retinal cell death and immunomodulation emerges as a therapeutic option. Indole-3-carbinol (I3C) is a natural ligand of aryl hydrocarbon receptor (AhR), with potent immunomodulatory properties. Here, we hypothesized that I3C may inhibit microglia reactivity and exert neuroprotective effects in the light-damaged murine retina mimicking important immunological aspects of retinal degeneration.

Methods

BV-2 microglia were treated in vitro with I3C followed by lipopolysaccharide (LPS) stimulation to analyze pro-inflammatory and anti-oxidant responses by quantitative real-time PCR (qRT-PCR) and Western blots. Nitric oxide (NO) secretion, caspase 3/7 levels, phagocytosis rates, migration, and morphology were analyzed in control and AhR knockdown cells. I3C or vehicle was systemically applied to light-treated BALB/cJ mice as an experimental model of retinal degeneration. Pro-inflammatory and anti-oxidant responses in the retina were examined by qRT-PCR, ELISA, and Western blots. Immunohistochemical staining of retinal flat mounts and cryosections were performed. The retinal thickness and structure were evaluated by in vivo imaging using spectral domain-optical coherence tomography (SD-OCT).

Results

The in vitro data showed that I3C potently diminished LPS-induced pro-inflammatory gene expression of I-NOS, IL-1ß, NLRP3, IL-6, and CCL2 and induced anti-oxidants gene levels of NQO1, HMOX1, and CAT1 in BV-2 cells. I3C also reduced LPS-induced NO secretion, phagocytosis, and migration as important functional microglia parameters. siRNA-mediated knockdown of AhR partially prevented the previously observed gene regulatory events. The in vivo experiments revealed that I3C treatment diminished light-damage induced I-NOS, IL-1ß, NLRP3, IL-6, and CCL2 transcripts and also reduced CCL2, I-NOS, IL-1ß, p-NFkBp65 protein levels in mice. Moreover, I3C increased anti-oxidant NQO1 and HMOX1 protein levels in light-exposed retinas. Finally, I3C therapy prevented the accumulation of amoeboid microglia in the subretinal space and protected from retinal degeneration.

Conclusions

The AhR ligand I3C potently counter-acts microgliosis and light-induced retinal damage, highlighting a potential treatment concept for retinal degeneration.
Literature
1.
Akhtar-Schäfer I, Wang L, Krohne TU, Xu H, Langmann T. Modulation of three key innate immune pathways for the most common retinal degenerative diseases. EMBO Mol Med. 2018;10:e8259.CrossRefPubMedPubMedCentral
2.
3.
4.
Karlstetter M, Scholz R, Rutar M, Wong WT, Provis JM, Langmann T. Retinal microglia: just bystander or target for therapy? Progress in Retinal and Eye Research. 2015;45:30–57.CrossRefPubMed
5.
Hao N, Whitelaw ML. The emerging roles of AhR in physiology and immunity. Biochem Pharmacol. 2013;86:561–70.PubMed
6.
7.
Bravo-Ferrer I, Cuartero MI, Medina V, Ahedo-Quero D, Peña-Martinez C, Pérez-Ruíz A, et al. Lack of the aryl hydrocarbon receptor accelerates aging in mice. FASEB J. 2019;33:12644–54.PubMed
8.
Brinkmann V, Ale-Agha N, Haendeler J, Ventura N. The aryl hydrocarbon receptor (AhR) in the aging process: another puzzling role for this highly conserved transcription factor. Front Physiol. 2020;10:1561.PubMedPubMedCentral
9.
Stockinger B, Di Meglio P, Gialitakis M, Duarte JH. The aryl hydrocarbon receptor: multitasking in the immune system. Ann Rev Immunol. 2014;32:403–32.
10.
Shinde R, McGaha TL. The aryl hydrocarbon receptor: connecting immunity to the microenvironment. Trends Immunol. 2018;39:1005–20.PubMedPubMedCentral
11.
Huang Y, He J, Liang H, Hu K, Jiang S, Yang L, et al. Aryl hydrocarbon receptor regulates apoptosis and inflammation in a murine model of experimental autoimmune uveitis. Front Immunol. 2018;9:1713.PubMedPubMedCentral
12.
Hu P, Herrmann R, Bednar A, Saloupis P, Dwyer MA, Yang P, et al. Aryl hydrocarbon receptor deficiency causes dysregulated cellular matrix metabolism and age-related macular degeneration-like pathology. Proc Natl Acad Sci U S A. 2013;110:E4069–78.PubMedPubMedCentral
13.
Kim SY, Chang YS, Chang YS, Kim JW, Brooks M, Chew EY, et al. Deletion of aryl hydrocarbon receptor AHR in mice leads to subretinal accumulation of microglia and RPE atrophy. Investig Ophthalmol Vis Sci. 2014;55:6031–40.
14.
Choudhary M, Kazmin D, Hu P, Thomas RS, McDonnell DP, Malek G. Aryl hydrocarbon receptor knock-out exacerbates choroidal neovascularization via multiple pathogenic pathways. J Pathol. 2015;235:101–12.PubMed
15.
Gutierrez MA, Davis SS, Rosko A, Nguyen SM, Mitchell KP, Mateen S, et al. A novel AhR ligand, 2AI, protects the retina from environmental stress. Sci Rep. 2016;6:29025.PubMedPubMedCentral
16.
Blasi E, Barluzzi R, Bocchini V, Mazzolla R, Bistoni F. Immortalization of murine microglial cells by a v-raf/v-myc carrying retrovirus. J Neuroimmunol. 1990;27:229–237.
17.
Scholz R, Sobotka M, Caramoy A, Stempfl T, Moehle C, Langmann T. Minocycline counter-regulates pro-inflammatory microglia responses in the retina and protects from degeneration. J Neuroinflammation. 2015;12:209.PubMedPubMedCentral
18.
Stansley B, Post J, Hensley K. A comparative review of cell culture systems for the study of microglial biology in Alzheimer’s disease. J Neuroinflammation. 2012;9:115.PubMedPubMedCentral
19.
Henn A, Lund S, Hedtjärn M, Schrattenholz A, Pörzgen P, Leist M. The suitability of BV2 cells as alternative model system for primary microglia cultures or for animal experiments examining brain inflammation. Altex. 2009;26:83–94.PubMed
20.
Majerova P, Zilkova M, Kazmerova Z, Kovac A, Paholikova K, Kovacech B, et al. Microglia display modest phagocytic capacity for extracellular tau oligomers. J Neuroinflammation. 2014;11:161.PubMedPubMedCentral
21.
Karlstetter M, Nothdurfter C, Aslanidis A, Moeller K, Horn F, Scholz R, et al. Translocator protein (18 kDa) (TSPO) is expressed in reactive retinal microglia and modulates microglial inflammation and phagocytosis. J Neuroinflammation. 2014;11:3.PubMedPubMedCentral
22.
Orihuela R, McPherson CA, Harry GJ. Microglial M1/M2 polarization and metabolic states. Br J Pharmacol. 2016;173:649–65.PubMed
23.
Lin H, Gao X, Chen G, Sun J, Chu J, Jing K, et al. Indole-3-carbinol as inhibitors of glucocorticoid-induced apoptosis in osteoblastic cells through blocking ROS-mediated Nrf2 pathway. Biochem Biophys Res Commun. 2015;460:422–7.PubMed
24.
Jiang J, Kang TB, Shim DW, Oh NH, Kim TJ, Lee KH. Indole-3-carbinol inhibits LPS-induced inflammatory response by blocking TRIF-dependent signaling pathway in macrophages. Food Chem Toxicol. 2013;57:256–61.PubMed
25.
Vorontsova JE, Cherezov RO, Kuzin BA, Simonova OB. Aryl-hydrocarbon receptor as a potential target for anticancer therapy. Biochem Suppl Ser B Biomed Chem. 2019;13:36–54.
26.
Karimabad MN, Mahmoodi M, Jafarzadeh A, Darekordi A, Hajizadeh MR, Hassanshahi G. Molecular targets, anti-cancer properties and potency of synthetic indole-3-carbinol derivatives. Mini-Rev Med Chem. 2018;19:540–54.
27.
Wang ML, Shih CK, Chang HP, Chen YH. Antiangiogenic activity of indole-3-carbinol in endothelial cells stimulated with activated macrophages. Food Chem. 2012;134:811–20.PubMed
28.
Tan E, Ding XQ, Saadi A, Agarwal N, Naash MI, Al-Ubaidi MR. Expression of cone-photoreceptor-specific antigens in a cell line derived from retinal tumors in transgenic mice. Investig Ophthalmol Vis Sci. 2004;45:764–8.
29.
Wheway G, Nazlamova L, Turner D, Cross S. 661W photoreceptor cell line as a cell model for studying retinal ciliopathies. Front Genet. 2019;10:308.PubMedPubMedCentral
30.
Sayyad Z, Sirohi K, Radha V, Swarup G. 661W is a retinal ganglion precursor-like cell line in which glaucoma-associated optineurin mutants induce cell death selectively. Sci Rep. 2017;7:1–13.CrossRef
31.
Wiedemann J, Rashid K, Langmann T. Resveratrol induces dynamic changes to the microglia transcriptome, inhibiting inflammatory pathways and protecting against microglia-mediated photoreceptor apoptosis. Biochem Biophys Res Commun. 2018;501:239–45.CrossRefPubMed
32.
Nomura K, Vilalta A, Allendorf DH, Hornik TC, Brown GC. Activated microglia desialylate and phagocytose cells via neuraminidase, galectin-3, and mer tyrosine kinase. J Immunol. 2017;198:4792–801.CrossRefPubMedPubMedCentral
33.
34.
Aslanidis A, Karlstetter M, Scholz R, Fauser S, Neumann H, Fried C, et al. Activated microglia/macrophage whey acidic protein (AMWAP) inhibits NFΚB signaling and induces a neuroprotective phenotype in microglia. J Neuroinflammation. 2015;12:77.CrossRefPubMedPubMedCentral
35.
Lu HF, Tung WL, Yang JS, Huang FM, Lee CS, Huang YP, et al. In vitro suppression of growth of murine WEHI-3 leukemia cells and in vivo promotion of phagocytosis in a leukemia mice model by indole-3-carbinol. J Agric Food Chem. 2012;60:7634–43.CrossRefPubMed
36.
Rico-Leo EM, Alvarez-Barrientos A, Fernandez-Salguero PM. Dioxin receptor expression inhibits basal and transforming growth factor β-induced epithelial-to-mesenchymal transition. J Biol Chem. 2013;288:7841–56.CrossRefPubMedPubMedCentral
37.
Josyula N, Andersen ME, Kaminski NE, Dere E, Zacharewski TR, Bhattacharya S. Gene co-regulation and co-expression in the aryl hydrocarbon receptor-mediated transcriptional regulatory network in the mouse liver. Arch Toxicol. 2020;94:113–26.CrossRefPubMed
38.
Huai W, Zhao R, Song H, Zhao J, Zhang L, Zhang L, et al. Aryl hydrocarbon receptor negatively regulates NLRP3 inflammasome activity by inhibiting NLRP3 transcription. Nat Commun. 2014;5:4738.CrossRefPubMed
39.
Wenzel A, Grimm C, Samardzija M, Remé CE. Molecular mechanisms of light-induced photoreceptor apoptosis and neuroprotection for retinal degeneration. Progress in Retinal and Eye Research. 2005;24:275–306.CrossRefPubMed
40.
Grimm C, Wenzel A, Stanescu D, Samardzija M, Hotop S, Groszer M, et al. Constitutive overexpression of human erythropoietin protects the mouse retina against induced but not inherited retinal degeneration. J Neurosci. 2004;24:5651–8.PubMedPubMedCentral
41.
Ampofo E, Lachnitt N, Rudzitis-Auth J, Schmitt BM, Menger MD, Laschke MW. Indole-3-carbinol is a potent inhibitor of ischemia–reperfusion–induced inflammation. J Surg Res. 2017;215:34–46.PubMed
42.
Paliwal P, Chauhan G, Gautam D, Dash D, Patne SCU, Krishnamurthy S. Indole-3-carbinol improves neurobehavioral symptoms in a cerebral ischemic stroke model. Naunyn Schmiedebergs Arch Pharmacol. 2018;391:613–25.PubMed
43.
Rutar M, Natoli R, Valter K, Provis JM. Early focal expression of the chemokine Ccl2 by Müller cells during exposure to damage-inducing bright continuous light. Investig Ophthalmol Vis Sci. 2011;52:2379–88.
44.
Rutar M, Natoli R, Provis JM. Small interfering RNA-mediated suppression of Ccl2 in Müller cells attenuates microglial recruitment and photoreceptor death following retinal degeneration. J Neuroinflammation. 2012;9:221.PubMedPubMedCentral
45.
Rothhammer V, Borucki DM, Tjon EC, Takenaka MC, Chao CC, Ardura-Fabregat A, et al. Microglial control of astrocytes in response to microbial metabolites. Nature. 2018;557:724–8.PubMedPubMedCentral
46.
Crunkhorn S. Autoimmune disease: aryl hydrocarbon receptor suppresses inflammation. Nat Rev Drug Discov. 2018;17:470.PubMed
47.
Higgins PJ. Balancing AhR-dependent pro-oxidant and Nrf2-responsive anti-oxidant pathways in age-related retinopathy: is SERPINE1 expression a therapeutic target in disease onset and progression? J Mol Genet Med. 2015;8:101.
48.
Perepechaeva ML, Grishanova AY, Rudnitskaya EA, Kolosova NG. The mitochondria-targeted antioxidant SkQ1 downregulates aryl hydrocarbon receptor-dependent genes in the retina of OXYS rats with AMD-like retinopathy. J Ophthalmol. 2014;2014:530943.PubMedPubMedCentral
49.
El-Naga RN, Ahmed HI, Abd Al Haleem EN. Effects of indole-3-carbinol on clonidine-induced neurotoxicity in rats: impact on oxidative stress, inflammation, apoptosis and monoamine levels. Neurotoxicology. 2014;44:48–57.PubMed
50.
Saini N, Akhtar A, Chauhan M, Dhingra N, Pilkhwal Sah S. Protective effect of indole-3-carbinol, an NF-κB inhibitor in experimental paradigm of Parkinson’s disease: In silico and in vivo studies. Brain Behav Immun. 2020;90:108–37.
51.
Anderton MJ, Manson MM, Verschoyle RD, Gescher A, Lamb JH, Farmer PB, et al. Pharmacokinetics and tissue disposition of indole-3-carbinol and its acid condensation products after oral administration to mice. Clin Cancer Res. 2004;10:5233–41.PubMed
52.
Juricek L, Carcaud J, Pelhaitre A, Riday TT, Chevallier A, Lanzini J, et al. AhR-deficiency as a cause of demyelinating disease and inflammation. Sci Rep. 2017;7:9794.PubMedPubMedCentral
53.
Rothhammer V, Quintana FJ. The aryl hydrocarbon receptor: an environmental sensor integrating immune responses in health and disease. Nature Reviews Immunology. 2019;19:184–97.PubMed
Metadata
Title
Indole-3-carbinol regulates microglia homeostasis and protects the retina from degeneration
Authors
Amir Saeed Khan
Thomas Langmann
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Journal of Neuroinflammation / Issue 1/2020
Electronic ISSN: 1742-2094
DOI
https://doi.org/10.1186/s12974-020-01999-8

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