Local inflammatory process as a possible manifestation of intraocular fluid uveolymphatic outflow defects in glaucoma. Part 2

PURPOSE: To study the ultrastructure of human trabecular zone by electron and light microscopy and inflammatory cytokines in intraocular fluid in primary open-angle glaucoma (POAG). METHODS: Fragments of eye tissues enucleated for medical indications (n=28) were studied. The main group included 17 eyes of patients diagnosed with terminal stage primary open-angle glaucoma. Obtained eye tissues were studied by means of Leica DME light microscope. Electron microscopy of the eye tissues was carried out using JEM 1400 electron microscope (Japan). 45 samples of intraocular fluid from patients with advanced POAG and 30 samples from patients with uncomplicated cataract were studied. Concentrations of 17 cytokines were detected by flow fluorimetry in double-beam laser analyzer (Bio-Plex 200, «Bio-Rad», USA) using Bio-Plex Pro™ Human Cytokine 17-plex Assay set («Bio-Rad», USA). RESULTS: Inflammatory cytokines IL-6, IL-8, and IL-17 were found to be significantly and interdependently increased in intraocular fluid of patients with advanced POAG. These results allow making a conclusion on the importance of local inflammation in POAG pathogenesis. Morphological study using light and electron microscopy detected the following changes in trabecular zone and Schlemm’s canal in terminal POAG: inflammatory infiltration, destruction, multidirectionality, swelling and merging of connected tissue fibers, a big amount of extracellular material in the basal membrane of Schlemm’s canal wall, an increase of lysosomes number, mitochondrial swelling in endothelium, and inter-endothelial connections density increase. These results could be considered as evidence of local inflammation and destructive processes. CONCLUSION: The study provided new basic science data obtained by light and electron microscopy, as well as multiplex assay, helps expand modern understanding of the role of local inflammation in POAG pathogenesis.

cytokines, aqueous humor, trabecular meshwork, inflammation, primary open-angle glaucoma

1. Borodin Yu.I., Bgatova N.P., Chernykh V.V., Trunov A.N. et al. Structure of lymphatic capillaries of the ciliary body of the human eye. Morphology. 2015; 148:43-47. (In Russ.).

2. Chernykh V.V., Borodin Yu.I., Bgatova N.P., Trunov A.N. et al. The role of the lymphatic system in the uveoscleral outflow of the intraocular fluid. Ophthalmosurgery. 2015; 2:74-79. (In Russ.).

3. Chernykh V.V., Borodin Yu.I., Bgatova N.P., Trunov A.N. et al. Structural elements of the lymphatic drainage channels of the intraocular fluid in the human choroid of the eye in normal and with glaucoma. Ophthalmosurgery. 2016; 3:11-16. (In Russ.).

4. Borodin Yu.I., Konenkov V.I., Lyubarsky M.S. Lymphology. Novosibirsk; 2011: 566. (In Russ.).

5. Chaudhry H.A., Dueker D.K., Simmons R.J., Bellows A.R., Grant W.M. Scanning electron microscopy of trabeculectomy specimens in openangle glaucoma. Am J Ophthalmol. 1979; 88(1):78-92.

6. Carreon T., van der Merwe E., Fellman R.L., Johnstone M., Bhattacharya S.K. Aqueous outflow - A continuum from trabecular meshwork to episcleral veins. Prog Retin Eye Res. 2017; 57:108-133. doi: 10.1016/j.preteyeres.2016.12.004.

7. Huang A.S., Mohindroo C., Weinreb R.N. Aqueous humor outflow structure and function imaging at the bench and bedside: a review. J Clin Exp Ophthalmol. 2016; 7(4):578. doi: 10.4172/2155-9570.1000578.

8. Lee N.Y., Park H.Y., Park C.K., Ahn M.D. Analysis of systemic endothelin-1, matrix metalloproteinase-9, macrophage chemoattractant protein-1, and high-sensitivity C-reactive protein in normal-tension glaucoma. Curr Eye Res. 2012; 37(12):1121-1126. doi: 10.3109/02713683.2012.725798.

9. Junglas B., Kuespert S., Seleem A.A., Struller T., Ullmann S., Bösl M., Bosserhoff A., Köstler J., Wagner R., Tamm E.R., Fuchshofer R. Connective tissue growth factor causes glaucoma by modifying the actin cytoskeleton of the trabecular meshwork. Am J Pathol. 2012; 180(6):2386-2403. doi: 10.1016/j.ajpath.2012.02.030.

10. Sihota R., Goyal A., Kaur J., Gupta V., Nag T.C. Scanning electron microscopy of the trabecular meshwork: understanding the pathogenesis of primary angle closure glaucoma. Indian J Ophthalmol. 2012; 60(3):183-188. doi: 10.4103/0301-4738.95868.

11. Song M.M., Lei Y., Wu J.H., Sun X.H. The progress of studies on aqueous humor dynamics abnormality induced by trabecular meshwork and Schlemm canal endothelial cell senescence and its relation with glaucoma. Zhonghua Yan Ke Za Zhi. 2017; 53(11):868-873. doi: 10.3760/cma.j.issn.0412-4081.2017.11.014.

12. Taurone S., Ripandelli G., Pacella E., Bianchi E. et al. Potential regulatory molecules in the human trabecular meshwork of patients with glaucoma: immunohistochemical profile of a number of inflammatory cytokines. Mol Med Rep. 2015; 11(2):1384-1390. doi: 10.3892/mmr.2014.2772.

13. Wang K., Read A.T., Sulchek T., Ethier C.R. Trabecular meshwork stiffness in glaucoma. Exp Eye Res. 2017; 158:3-12. doi: 10.1016/j.exer.2016.07.011.

14. Chernykh V.V. Effect of complex conservative therapy on immunobiochemical indices in tear fluid in patients with primary open-angle glaucoma. Glaucoma. 2005; 4:20-24. (In Russ.).

15. Chernykh V.V., Shvyak A.P., Gorbenko O.M., Safronov I.D., et al. Peculiarities of the pathogenesis of the initial and advanced stage of primary open-angle glaucoma. Allergology and Immunology. 2006; 1:28-31. (In Russ.).

16. Trunov A.N., Bgatova N.P., Eremina A.V., Druzhinin I.B. et al. New approaches to assessing the severity of inflammatory disorders in the pathogenesis of primary open-angle glaucoma. Allergology and Immunology. 2016; 17(2):107-111. (In Russ.).

17. Erichev V.P., Gankovskaya L.V., Kovalchuk L.V., Gankovskaya O.A. et al. Changes in some immunological parameters of lacrimal fluid with excessive scarring after antiglaucomatous operations in patients with primary open-angle glaucoma. Vestn oftalmol. 2010; 126(3): 25-29. (In Russ.).

18. Khokhlova A.S., Kirienko A.V., Filina N.V., Markelova E.V. Local cytokine regulation at different stages of primary open-angle glaucoma. Pacific Medical Journal. 2014; 4:46-48. (In Russ.).

19. Cherednichenko L.P., Barycheva L.Yu., Bernovskaya A.A. Determination of proinflammatory cytokines in early diagnosis of primary openangle glaucoma. Russian Ophthalmological Journal. 2013; 6(2):82-85. (In Russ.).

20. Benitez-Del-Castillo Sánchez J., Morillo-Rojas M.D., Galbis-Estrada C., Pinazo-Duran M.D. Determination of inmune response and inflammation mediators in tears: Changes in dry eye and glaucoma as compared to healthy controls. Arch Soc Esp Oftalmol. 2017; 92(5): 210-217. doi: 10.1016/j.oftal.2016.12.009.

21. Chernyavskaya M.A., Efremov A.V., Chernykh V.V. Interrelation of cytokines in the lacrimal and intraocular fluid with melanoma of the choroid. Bulletin of Tambov University. Series: Natural and technical sciences. 2015; 20(3):710-712. (In Russ.).

22. Shpak A.A., Gekht A.B., Druzhkova T.A., Kozlova K.I., Gulyaeva N.V. Ratios of neurotrophic factors in tear fluid and moisture in the anterior chamber in patients with age-related cataract. Ophthalmosurgery. 2017; 1:16-20. (In Russ.).

23. Chua J., Vania M., Cheung C.M., Ang M., Chee S.P. et al. Expression profile of inflammatory cytokines in aqueous from glaucomatous eyes. Mol Vis. 2012; 18:431-438.

24. Kokubun T., Tsuda S., Kunikata H., Yasuda M. et al. Characteristic profiles of inflammatory cytokines in the aqueous humor of glaucomatous eyes. Ocul Immunol Inflamm. 2017; 16:1-12. doi: 10.1080/09273948.2017.1327605.

25. Duvesh R., Puthuran G., Srinivasan K., Rengaraj V. et al. Multiplex cytokine analysis of aqueous humor from the patients with chronic primary angle closure glaucoma. Curr Eye Res. 2017; 42(12):1608- 1613. doi: 10.1080/02713683.2017.1362003.

26. Tong Y., Zhou Y.L., Zheng Y., Biswal M. et al. Analyzing cytokines as biomarkers to evaluate severity of glaucoma. Int J Ophthalmol. 2017; 10(6):925-930. doi: 10.18240/ijo.2017.06.15.