TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium

Makoto Shirasawa, Shozo Sonoda, Hiroto Terasaki, Noboru Arimura, Hiroki Otsuka, Takehiro Yamashita, Eisuke Uchino, Toshio Hisatomi, Tatsuro Ishibashi, Taiji Sakamoto

Research output: Contribution to journalArticle

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Abstract

Retinal pigment epithelial (RPE) cells form a blood-ocular barrier, and their polarized property is crucial for maintaining the barrier functions. Tumor necrosis factor alpha (TNF-α), a major pleotropic inflammatory cytokine that disrupts the barrier function and eventual angiogenesis, is expressed in the choroidal neovascularizations of age-related macular degeneration eyes. Thus, it most likely plays an important role in the progression of the disease. The purpose of this study was to compare the effects of TNF-α on the barrier function of polarized RPE cells. Non-polarized RPE cells were used as negative controls. Isolated porcine RPE cells were seeded on Transwell™ membranes. The polarization of the RPE cells was determined by their high transepithelial electrical resistance (TER >150 Ω cm2) and by their differential secretion of vascular endothelial growth factor (lower layer/upper layer >2.5X). Polarized RPE cells were incubated with 10 ng/ml of TNF-α and the TER was measured. TNF-α significantly decreased the TER of polarized RPE cells by 17.6 ± 2.7% (P < 0.001) of the control at 24 h and that of non-polarized RPE cells by 5.4 ± 6.5% (P = 0.401). The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked the effects of TNF-α of decreasing the TER. Cell junction-related molecules, e.g., ZO-1, located between cells in control RPE cells, were disassembled by TNF-α, and this breakdown was suppressed by SB203580 in polarized RPEs. These results indicate that the breakdown of the RPE barrier function was caused exclusively by TNF-α in polarized RPEs, and TNF-α was acting through the p38 MAPK pathways. Investigations of polarized RPE cells should be more suitable for in vitro studies of the pathophysiology of retinochoroidal diseases.

Original languageEnglish
Pages (from-to)59-69
Number of pages11
JournalExperimental Eye Research
Volume110
DOIs
Publication statusPublished - May 1 2013

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Retinal Pigments
Retinal Pigment Epithelium
Tumor Necrosis Factor-alpha
Epithelial Cells
p38 Mitogen-Activated Protein Kinases
Choroidal Neovascularization
Intercellular Junctions
Macular Degeneration
Protein Kinase Inhibitors
Electric Impedance
Vascular Endothelial Growth Factor A
Disease Progression
Swine
Cytokines

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Shirasawa, M., Sonoda, S., Terasaki, H., Arimura, N., Otsuka, H., Yamashita, T., ... Sakamoto, T. (2013). TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium. Experimental Eye Research, 110, 59-69. https://doi.org/10.1016/j.exer.2013.02.012

TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium. / Shirasawa, Makoto; Sonoda, Shozo; Terasaki, Hiroto; Arimura, Noboru; Otsuka, Hiroki; Yamashita, Takehiro; Uchino, Eisuke; Hisatomi, Toshio; Ishibashi, Tatsuro; Sakamoto, Taiji.

In: Experimental Eye Research, Vol. 110, 01.05.2013, p. 59-69.

Research output: Contribution to journalArticle

Shirasawa, M, Sonoda, S, Terasaki, H, Arimura, N, Otsuka, H, Yamashita, T, Uchino, E, Hisatomi, T, Ishibashi, T & Sakamoto, T 2013, 'TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium', Experimental Eye Research, vol. 110, pp. 59-69. https://doi.org/10.1016/j.exer.2013.02.012
Shirasawa, Makoto ; Sonoda, Shozo ; Terasaki, Hiroto ; Arimura, Noboru ; Otsuka, Hiroki ; Yamashita, Takehiro ; Uchino, Eisuke ; Hisatomi, Toshio ; Ishibashi, Tatsuro ; Sakamoto, Taiji. / TNF-α disrupts morphologic and functional barrier properties of polarized retinal pigment epithelium. In: Experimental Eye Research. 2013 ; Vol. 110. pp. 59-69.
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abstract = "Retinal pigment epithelial (RPE) cells form a blood-ocular barrier, and their polarized property is crucial for maintaining the barrier functions. Tumor necrosis factor alpha (TNF-α), a major pleotropic inflammatory cytokine that disrupts the barrier function and eventual angiogenesis, is expressed in the choroidal neovascularizations of age-related macular degeneration eyes. Thus, it most likely plays an important role in the progression of the disease. The purpose of this study was to compare the effects of TNF-α on the barrier function of polarized RPE cells. Non-polarized RPE cells were used as negative controls. Isolated porcine RPE cells were seeded on Transwell™ membranes. The polarization of the RPE cells was determined by their high transepithelial electrical resistance (TER >150 Ω cm2) and by their differential secretion of vascular endothelial growth factor (lower layer/upper layer >2.5X). Polarized RPE cells were incubated with 10 ng/ml of TNF-α and the TER was measured. TNF-α significantly decreased the TER of polarized RPE cells by 17.6 ± 2.7{\%} (P < 0.001) of the control at 24 h and that of non-polarized RPE cells by 5.4 ± 6.5{\%} (P = 0.401). The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked the effects of TNF-α of decreasing the TER. Cell junction-related molecules, e.g., ZO-1, located between cells in control RPE cells, were disassembled by TNF-α, and this breakdown was suppressed by SB203580 in polarized RPEs. These results indicate that the breakdown of the RPE barrier function was caused exclusively by TNF-α in polarized RPEs, and TNF-α was acting through the p38 MAPK pathways. Investigations of polarized RPE cells should be more suitable for in vitro studies of the pathophysiology of retinochoroidal diseases.",
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