2-ethylpyridine, a cigarette smoke component, causes mitochondrial damage in human retinal pigment epithelial cells in vitro.
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Date
2014-01
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Abstract
Purpose: Our goal was to identify the cellular and molecular effects of 2-ethylpyridine (2-EP,
a component of cigarette smoke) on human retinal pigment epithelial cells (ARPE-19) in vitro.
Materials and Methods: ARPE-19 cells were exposed to varying concentrations of 2-EP. Cell viability (CV)
was measured by a trypan blue dye exclusion assay. Caspase-3/7 and caspase-9 activities were measured
by fl uorochrome assays. The production of reactive oxygen/nitrogen species (ROS/RNS) was detected with
a 2’,7’-dichlorodihydrofl uorescein diacetate dye assay. The JC-1 assay was used to measure mitochondrial
membrane potential (m). Mitochondrial redox potential was measured using a RedoxSensor Red kit
and mitochondria were evaluated with Mitotracker dye. Results: Aft er 2-EP exposure, ARPE-19 cells
showed signifi cantly decreased CV, increased caspase-3/7 and caspase-9 activities, elevated ROS/RNS
levels, decreased m value and decreased redox fl uorescence when compared with control samples.
Conclusions: These results show that 2-EP treatment induced cell death by caspase-dependent apoptosis
associated with an oxidative stress and mitochondrial dysfunction. These data represent a possible
mechanism by which smoking contributes to age-related macular degeneration and other retinal diseases
and identify mitochondria as a target for future therapeutic interventions.
Description
Keywords
2-ethylpyridine, apoptosis, ARPE-19 cells, cigarett e smoke toxicant, mitochondrial membrane potential, reactive oxygen/nitrogen species
Citation
Mansoor S, Gupta N, Falatoonzadeh P, Kuppermann B D, Kenney M C. 2-ethylpyridine, a cigarette smoke component, causes mitochondrial damage in human retinal pigment epithelial cells in vitro. Indian Journal of Ophthalmology. 2014 Jan ; 62 (1): 16-22.