Increased stress-induced generation of reactive oxygen species and apoptosis in human keratoconus fibroblasts

Investigative Ophthalmology & Visual Science
Marilyn ChwaM Cristina Kenney

Abstract

To determine whether keratoconus (KC) corneal fibroblast cultures have increased reactive oxygen species (ROS) production and are more susceptible to stress-related challenges. Normal (n = 9) and KC (n = 10) stromal fibroblast cultures were incubated in either neutral- or low-pH conditions, with or without hydrogen peroxide. Catalase activities were measured with a fluorescent substrate assay. Superoxide and ROS/reactive nitrogen species (RNS) productions were determined with an amine-reactive green-dye assay and 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) dye assay, respectively. Cell viability was analyzed by a dye-exclusion assay. Caspase 3 activity was measured by a fluorochrome inhibitor of caspase (FLICA) assay. A cationic (green) dye was used to measure the mitochondrial membrane potential (delta psi m). KC fibroblasts had increased superoxide and ROS/RNS production (6.2-fold, P < 0.001 and 1.8-fold, P < 0.001, respectively) and catalase activity (P < 0.01) with higher concentrations of H2O2 compared with normal cultures (P = 0.16). After a low-pH stress challenge, KC fibroblasts maintained higher ROS/RNS levels (3.3-fold, P < 0.02), showed higher caspase-3 activity (7.5-fold, P < 0.02) and decreased delta psi m...Continue Reading

Citations

May 5, 2009·Cornea·Shari R AtilanoM Cristina Kenney
Aug 6, 2009·Cornea·Nitin UdarM Cristina Kenney
Aug 31, 2010·Cornea·Arie Y NemetIgor Kaiserman
Feb 8, 2011·PloS One·Albert S JunShukti Chakravarti
Sep 26, 2013·International Journal of Molecular Sciences·Katarzyna A WojcikJacek P Szaflik
Feb 7, 2015·Current Eye Research·Raşit KılıçHafize Sezer
Apr 20, 2011·Current Eye Research·Luisa Simo MannionClare O'Donnell
Oct 31, 2012·Tissue & Cell·Cheng-Ye WangZhong-Bao Zhao
Sep 11, 2012·Experimental Eye Research·Aluru Venkata SaijyothiNarayanasamy Angayarkanni
Jul 28, 2007·Experimental Eye Research·Steven E WilsonFabricio W Medeiros
Dec 2, 2015·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Claudio RamírezM Cristina Kenney
Jan 23, 2013·Clinical & Experimental Optometry : Journal of the Australian Optometrical Association·Isabella M Y CheungTrevor Sherwin
Feb 18, 2014·Clinical & Experimental Optometry : Journal of the Australian Optometrical Association·Isabella M Y CheungTrevor Sherwin
May 2, 2015·Eye·V GalvisA Acera
Sep 26, 2014·Journal of Ophthalmology·Khaled K Abu-AmeroAltaf A Kondkar
Jul 27, 2014·International Ophthalmology·Rana SorkhabiMohammad Hosein Ahoor
Nov 3, 2016·Zygote : the Biology of Gametes and Early Embryos·A L S GuimarãesM A N Dode
Jan 7, 2017·Experimental Biology and Medicine·Tina B McKay, Dimitrios Karamichos
Feb 18, 2017·Scientific Reports·Tina B McKayDimitrios Karamichos
Sep 30, 2017·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Mohamed Tarek MoustafaMaria Cristina Kenney
Jun 6, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Katerina KuzelováZbynek Hrkal
Apr 5, 2019·Eye & Contact Lens·Döndü Melek UlusoyAyşe Çiçek
Jun 8, 2012·The Journal of Reproduction and Development·K M A TareqHirotada Tsujii

❮ Previous
Next ❯

Related Concepts

Related Feeds

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis

Apoptotic Caspases

Apoptotic caspases belong to the protease enzyme family and are known to play an essential role in inflammation and programmed cell death. Here is the latest research.