Nonlethal Oxidant Injury to Human Retinal Pigment Epithelium Cells Causes Cell Membrane Blebbing but Decreased MMP-2 Activity

Marin‐Castaño, Maria E.; Csaky, Karl G.; Cousins, Scott W.

doi:10.1167/iovs.04-1224

Cited by 66 publications

(71 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…20 We previously found that exposure to cigarette smoke and HQ resulted in RPE membrane blebbing and sub-RPE deposits in mice. 21,22 We also demonstrated that nonlethal HQ-induced oxidative injury in cultured RPE cells results in reorganization of actin cytoskeleton and blebs formation [22][23][24][25] relevant to the accumulation of deposits. Blebbing of the plasma membrane is an early morphological sign of cell injury, which occurs immediately after exposure to a wide variety of toxic agents.…”

mentioning

confidence: 66%

“…24 GFP-ARPE-19 cells were pretreated for 1 hour with or without 20 mol/L of SB203580 or 60 nmol/L of okadaic acid, and then exposed to 100 mol/L of HQ for 6 hours. Figure 6, A and B, shows the extensive blebbing that occurs in GFP-ARPE-19 cells after exposure to nonlethal concentrations of HQ for 6 hours.…”

Section: Inhibition Of Hsp27 Phosphorylation Decreases Hq-induced Blementioning

confidence: 99%

See 1 more Smart Citation

Cigarette Smoke-Related Hydroquinone Induces Filamentous Actin Reorganization and Heat Shock Protein 27 Phosphorylation through p38 and Extracellular Signal-Regulated Kinase 1/2 in Retinal Pigment Epithelium

Pons

Cousins

Csaky

et al. 2010

The American Journal of Pathology

Self Cite

View full text Add to dashboard Cite

Retinal pigment epithelium (RPE)-derived membranous debris named blebs, may accumulate and contribute to sub-RPE deposit formation, which is the earliest sign of age-related macular degeneration (AMD). Oxidative injury to the RPE might play a significant role in AMD. However, the underlying mechanisms are unknown. We previously reported that hydroquinone (HQ), a major pro-oxidant in cigarette smoke, foodstuff, and atmospheric pollutants, induces actin rearrangement and membrane blebbing in RPE cells as well as sub-RPE deposits in mice. Here, we show for the first time that phosphorylated Heat shock protein 27 (Hsp27), a key regulator of actin filaments dynamics, is up-regulated in RPE from patients with AMD. Also, HQ-induced nonlethal oxidative injury led to Hsp27mRNA up-regulation, dimer formation , and Hsp27 phosphorylation in ARPE-19 cells. Furthermore, we found that a cross talk between p38 and extracellular signal-regulated kinase (ERK) mediates HQ-induced Hsp27 phosphorylation and actin aggregate formation, revealing ERK as a novel upstream mediator of Hsp27 phosphorylation. Finally, we demonstrated that Hsp25, p38, and ERK phosphorylation are increased in aging C57BL/6 mice chronically exposed to HQ, whereas Hsp25 expression is decreased. Our data suggest that phosphorylated Hsp27 might be a key mediator in AMD and HQ-induced oxidative injury to the RPE, which may provide helpful insights into the early cellular events associated with actin reorganization and bleb formation involved in sub-RPE deposits formation relevant to the pathogenesis of AMD.

show abstract

mentioning

confidence: 66%

Section: Inhibition Of Hsp27 Phosphorylation Decreases Hq-induced Blementioning

confidence: 99%

Cigarette Smoke-Related Hydroquinone Induces Filamentous Actin Reorganization and Heat Shock Protein 27 Phosphorylation through p38 and Extracellular Signal-Regulated Kinase 1/2 in Retinal Pigment Epithelium

Pons

Cousins

Csaky

et al. 2010

The American Journal of Pathology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Supporting the original reason why the eyecups were used to keep the RPE monolayer, the ARPE-19 cells grown to an established monolayer were able to suppress the activation of the phagolysosome in the phagocytizing macrophages. Other studies have grown the ARPE-19 cells to form an established monolayer before being assayed [8]. It demonstrates that as an intact monolayer the ARPE-19 cell line is very useful in understanding the role of human RPE produced factors in modulating immunity.…”

Section: Discussionmentioning

confidence: 99%

“…Since there are many reports showing almost the same activity of ARPE-19 cell monolayers to cultured primary RPE [8][9][10][11], the conditioned media of cultured ARPE-19 cells were assayed to see whether they suppress phagolysosome activation in phagocytizing macrophages. The ARPE-19 cells were cultured to confluence, or kept subconfluent before changing the media to the serum-free media for an additional 24 hours of incubation.…”

Section: The Effects Of Cultured Arpe-19 Cells On Macrophage Phagolysmentioning

confidence: 99%

“…One way around this limitation is to use the human RPE cell line ARPE-19. These cells grow to a confluent monolayer, and have been used to study the effects of oxidants and cytokines involved in RPE changes related to age-related macular degeneration [8,9]. These cell lines when treated with TGF-β2 have been found to mediate the activation of Treg cells [10].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Retinal Pigment Epithelial Cell Line Suppression of Phagolysosome Activation

Taylor

Dixit

2015

IJOES

View full text Add to dashboard Cite

The eye is an immune privileged tissue with multiple mechanisms of immunosuppression to protect the light gathering tissues from the damage of inflammation. One of theses mechanisms involves retinal pigment epithelial cell suppression of phagosome activation in macrophages. The objective of this work is to determine if the human RPE cell line ARPE-19 is capable of suppressing the activation of the phagolysosome in macrophages in a manner similar to primary RPE. The conditioned media of RPE eyecups, sub-confluent, just confluent cultures, or established confluent cultures of human ARPE-19 cells were generated. These condition media were used to treat macrophages phagocytizing pHrodo bioparticles. After 24 hours incubation the macrophages were imaged by fluorescent microscopy, and fluorescence was measured. The fluorescent intensity is proportional to the amount of bioparticles phagocytized and are in an activated phagolysosome. The conditioned media of in situ mouse RPE eyecups significantly suppressed the activation of phagolysosome. The conditioned media from cultures of human ARPE-19 cells, grown to sub-confluence (50%) or grown to confluence had no effect on phagolysosome activation. In contrast, the conditioned media from established confluent cultures significantly suppressed phagolysosome activation. The neuropeptides alpha-MSH and NPY were depleted from the conditioned media of established confluent ARPE-19 cell cultures. This depleted conditioned media had diminished suppression of phagolysosome activation while promoting macrophage cell death. In addition, the condition media from cultures of ARPE-19 monolayers wounded with a bisecting scrape was diminished in suppressing phagolysosome activation. This technical report suggests that like primary RPE monolayers, established confluent cultures of ARPE-19 cells produce soluble factors that suppress the activation of macrophages, and can be used to study the molecular mechanisms of retinal immunobiology. In addition, the results further demonstrate the importance of an intact monolayer of RPE cells to modulate immune cell activity within the eye.

show abstract

Smoking and Eye Diseases

Marin‐Castaño

Pons

2011

Cigarette Smoke Toxicity

View full text Add to dashboard Cite

Nonlethal Oxidant Injury to Human Retinal Pigment Epithelium Cells Causes Cell Membrane Blebbing but Decreased MMP-2 Activity

Cited by 66 publications

References 47 publications

Cigarette Smoke-Related Hydroquinone Induces Filamentous Actin Reorganization and Heat Shock Protein 27 Phosphorylation through p38 and Extracellular Signal-Regulated Kinase 1/2 in Retinal Pigment Epithelium

Cigarette Smoke-Related Hydroquinone Induces Filamentous Actin Reorganization and Heat Shock Protein 27 Phosphorylation through p38 and Extracellular Signal-Regulated Kinase 1/2 in Retinal Pigment Epithelium

Retinal Pigment Epithelial Cell Line Suppression of Phagolysosome Activation

Smoking and Eye Diseases

Contact Info

Product

Resources

About