期刊
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE
卷 62, 期 3, 页码 -出版社
ASSOC RESEARCH VISION OPHTHALMOLOGY INC
DOI: 10.1167/iovs.62.3.3
关键词
corneal endothelium; proteome; cigarette smoke
资金
- National Eye Institute [R01EY028538, R01EY027691]
- Margaret C. Mosher Professor of Ophthalmology
- Robert Bond Welch Professorship
- National Institutes of Health shared instrumentation grant
This study found that exposure to chronic cigarette smoke led to a decrease in corneal endothelial cell density in mice, accompanied by reduced levels of multiple collagen and extracellular matrix proteins associated with Descemet's membrane.
PURPOSE. To investigate changes at a molecular level in the mouse corneal endothelium (CE) exposed to chronic cigarette smoke (CS). METHODS. Pregnant mice (gestation days 18-20) were placed in a whole-body exposure smoking chamber, and a few days later pups were born. After 3.5 months of CS exposure, a ConfoScan4 scanning microscope was used to examine the corneal endothelial cells (CECs) of CS-exposed and control (Ct) mice. The CE was peeled under a microscope and maintained as four biological replicates (two male and two female) for CS-exposed and Ct mice; each replicate consisted of 16 CEs. The proteome of the CE was investigated through mass spectrometry. RESULTS. The CE images of CS-exposed and Ct mice revealed a difference in the shape of CECs accompanied by a nearly 10% decrease in CEC density (P < 0.00003) following CS exposure. Proteome profiling identified a total of 524 proteins exhibiting statistically significant changes in CE from CS-exposed mice. Importantly, proteins associated with Descemet's membrane (DM), including COL4 alpha 1, COL4 alpha 2, COL4 alpha 3, COL4 alpha 4, COL4 alpha 5, COL4 alpha 6, COL8 alpha 1, COL8 alpha 2, and FN1, among others, exhibited diminished protein levels in the CE of CS-exposed mice. CONCLUSIONS. Our data confirm that exposure to CS results in reduced CEC density accompanied by diminished levels of multiple collagen and extracellular matrix proteins associated with DM.
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