Features of local expression of mRNA, IL-1β, IL-18, CCL2/MCP-1 in modeling pigment epithelium atrophy and retinal degeneration in the experiment on rabbits

DOI: https://doi.org/10.29296/24999490-2021-02-08

V.V. Neroev, N.V. Balatskaya, E.V. Svetlova, N.V. Neroeva, M.V. Ryabina, A.G. Karmokova, O.A. Losanova, I.Yu. Chernomorets, P.A. Ilyukhin Helmholtz National Medical Research Center of Eye Diseases, Ministry of Health of Russia, Sadovaya-Chernogryazskaya, 14/19, Moscow, 105062, Russian Federation Е-mail: [email protected]

Introduction. Degenerative retinal diseases are considered as the leading cause of blindness and low vision worldwide. The last studies have provided convincing evidence of the involvement of immunological factors in the development and progression of the disease. Meanwhile the immunopathogenesis of the wet form of AMD has been studied more fully than the atrophic one. The aim of the study. Study of local expression of mRNA of inflammatory cytokines IL-1β, IL-18, CCL2/MCP-1 in a model of RPE atrophy. Material and methods. The material for the study was samples of the retinal-RPE-choroid (TC) tissue complex isolated at various times from the enucleated rabbits’ eyes with previously created RPE atrophy by subretinal injection of 0,9% sodium chloride solution. Optical coherence tomography (OCT) and autofluorescence (AF) study using HeidelbergSpectralis ™ SD-OCT were performed. Local expression of cytokines was determined by RT-PCR. Results. A single subretinal injection of 0.01 ml of 0.9% sodium chloride solution was found to be associated with multidirectional changes in the expression of mRNA of the IL-1β, IL-18, MCP-1/CCL2 genes relative to the norm. Three types of the retinal response, formed during the development of atrophic changes and determined by the degree of local mRNA expression of cytokine genes, were characterized. No correlation was found between the magnitude of local mRNA expression to the stimulus inducing atrophy and the size of the atrophic retinal lesion. Conclusion. Obtained data can be helpful in the research of RPE atrophy and in the stem cells treatment.
Keywords: 
atrophy of retinal pigment epithelium, retinal degeneration, tissue complex retina – choroid, inflammatory mediators, gene expression, RT-PCR in real time
Список литературы: 
  1. Machalińska A., Lubiński W., Kłos P., Kawa M., Baumert B., Penkala K., Grzegrzółka R., Karczewicz D., Wiszniewska B., Machaliński B. Sodium Iodate Selectively Injuries the Posterior Pole of the Retina in a Dose-Dependent Manner: Morphological and Electrophysiological Study. Neurochem. 2010; 35: 1819–27. https://doi.org/10.1007/s11064-010-0248-6
  2. Bhutto I., Lutty G. Understanding age-related macular degeneration (AMD): relationships between the photoreceptor/retinal pigment epithelium/Bruch's membrane/choriocapillaris complex. Mol Aspects Med. 2012; 33 (4): 295–317. http://dx.doi.org/10.1016/j.mam.2012.04.005
  3. Cunha-Vaz J., Bernardes R., Lobo C. Blood-retinal barrier. European J. of Ophthalmology. 2011; 21 (6): 3–9. https://doi.org/10.5301/EJO.2010.6049
  4. Oner A. Stem Cell Treatmentin Retinal Diseases: Recen Developments. Turkish J. ofophthalmology. 2018; 48 (1): 33–8. https://doi.org/10.4274/tjo.89972
  5. Kauppinen A., Paterno J.J., Blasiak J., Salminen A., Kaarniranta K. Inflammation and its role in age-related macular degeneration. Cellular and molecular life sciences. 2016; 73 (9): 1765–86. https://doi.org/10.1007/s00018-016-2147-8
  6. Whitcup S.M., Sodhi A., Atkinson J.P., Holers V.M., Sinha D., Rohrer B., Dick A.D. The role of the immune response in age-related macular degeneration. International J. of inflammation. 2013; 348092. https://doi.org/10.1155/2013/348092
  7. Doktor F., Prager P., Wiedemann P., Kohen L., Bringmann A., Hollborn M. Hypoxic expression of NLRP3 and VEGF in cultured retinal pigment epithelial cells: contribution of P2Y2 receptor signaling. Purinergic Signalling. 2018; 14 (4): 471–84. https://doi.org/10.1007/s11302-018-9631-6
  8. Kerur N., Hirano Y., Tarallo V., Fowler B.J., Bastos-Carvalho A., Yasuma T., Yasuma R., Kim Y., Hinton D.R., Kirschning C.J., Gelfand B.D., Ambati J. TLR-independent and P2X7-dependent signaling mediate Alu RNA-induced NLRP3 inflammasome activation in geographic atrophy. Invest Ophthalmol Vis Sci. 2013; 54 (12): 7395–401. https://doi.org/10.1007/978-3-319-17121-0_9
  9. Kauppinen A., Niskanen H., Suuronen T., Kinnunen K., Salminen A., Kaarniranta K. Oxidative stress activates NLRP3 inflammasomes in ARPE-19 cells – implications for age-related macular degeneration (AMD) Immunology Letters. 2012; 147 (1–2): 29–33. http://dx.doi.org/10.1016/j.imlet.2012.05.005
  10. Abe T., Sugano E., Saigo Y., Tamai M. Interleukin -1betta and barrier function of retinal pigment epithelial cells (ARPE-19): Abberant expression of junctional complex molecules. Investigative Ophthalmology & Visual Science. 2003; 44: 4097–104. http://dx.doi.org/10.1167/iovs.02-0867
  11. Tseng W.A., Then T., Kinnunen K., Lashkari K., Gregory M.S., D’Amore P.A., Ksander B.R. NLRP3 inflamasome activation in retinal pigment epithelial cells by lysosomal destabilization: Implication for age-related macular degeneration. Investigative ophthalmology & visual science. 2013; 54 (1): 110–20. https://doi.org/10.1167/iovs.12-10655
  12. Bamias G., Corridoni D., Pizzaro T.T., Cominelli F. New insights into the dichotomous role of innate cytokines in gut homeostasis and inflammation. Cytokine. 2012; 59: 451–9. http://dx.doi.org/10.1016/j.cyto.2012.06.014
  13. Bian Z.M., Elner S.H., Yoshida A., Kinkel S.L., Su J., Elner V.M. Activation of p38, ERK1/2 and NIK pathways is required for IL-1 betta and TNF-alpha-indused chemokine expression in human retinal pigment epithelial cells. Experimental Eye Research. 2001; 73: 111–21. https://doi.org/10.1006/exer.2001.1019
  14. Narimatsu T., Ozava Y., Miyake S., Hirasava M., Nagai N., Shimmura S., Tsubota K. Disruption of Cell-Cell Junctions and Induction of Pathological Cytokines in the Retinal Pigment Epithelium of Light-Exposed Mice. Investigative ophthalmology & visual science. 2013; 54: 4555–62. http://dx.doi.org/10.1167/iovs.12-11572
  15. Luheshi N.M., Kovács K.J., Lopez-Castejon G., Brough D., Denes A. Interleukin-1α expression precedes IL-1β after ischemic brain injury and is localised to areas of focal neuronal loss and penumbral tissues. J. of Neuroinflammation. 2011; 8: 186. http://dx.doi.org/10.1186/1742-2094-8-186