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Dynamic ocular surface and lacrimal gland changes induced in experimental murine dry eye.

Xiao B, Wang Y, Reinach PS, Ren Y, Li J, Hua S, Lu H, Chen W - PLoS ONE (2015)

Bottom Line: To determine if there is a correlation between severity effects in these models and underlying pathophysiological responses, we compared the time dependent changes in each of these parameters that occur during a 6 week period.Interleukin-17(IL-17), IL-23, IL-6, IL-1, TNF-α, IFN-γ and TGF-β2 levels were estimated by real-time PCR measurements of conjunctival and lacrimal gland samples (LGs).Subsequently, the disease process stabilized for the next four weeks.

View Article: PubMed Central - PubMed

Affiliation: School of Ophthalmology and Optometry, Wenzhou Medical University, Zhejiang, China.

ABSTRACT
Dry eye disease can be a consequence of lacrimal gland insufficiency in Sjögren's Syndrome or increased tear film evaporation despite normal lacrimal gland function. To determine if there is a correlation between severity effects in these models and underlying pathophysiological responses, we compared the time dependent changes in each of these parameters that occur during a 6 week period. Dry eye was induced in 6-week-old female C57BL/6 mice by exposing them to an Intelligently Controlled Environmental System (ICES). Sixty mice were housed in ICES for 1, 2, 4 and 6 weeks respectively. Twelve were raised in normal environment and received subcutaneous injections of scopolamine hydrobromide (SCOP) 3 times daily for 5 days. Another sixty mice were housed in a normal environment and received no treatment. Corneal fluorescein staining along with corneal MMP-9 and caspase-3 level measurements were performed in parallel with the TUNEL assay. Interleukin-17(IL-17), IL-23, IL-6, IL-1, TNF-α, IFN-γ and TGF-β2 levels were estimated by real-time PCR measurements of conjunctival and lacrimal gland samples (LGs). Immunohistochemistry of excised LGs along with flow cytometry in cervical lymph nodes evaluated immune cell infiltration. Light and transmission electron microscopy studies evaluated LGs cytoarchitectural changes. ICES induced corneal epithelial destruction and apoptosis peaked at 2 weeks and kept stable in the following 4 weeks. In the ICES group, lacrimal gland proinflammatory cytokine level increases were much lower than those in the SCOP group. In accord with the lower proinflammatory cytokine levels, in the ICES group, lacrimal gland cytosolic vesicular density and size exceeded that in the SCOP group. ICES and SCOP induced murine dry eye effects became progressively more severe over a two week period. Subsequently, the disease process stabilized for the next four weeks. ICES induced local effects in the ocular surface, but failed to elicit lacrimal gland inflammation and cytoarchitectural changes, which accounts for less dry eye severity in the ICES model than that in the SCOP model.

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ICES Induced Corneal Epithelium Apoptosis.A,ICES induced caspase-3 expression in each group. (green, caspase-3–positive; blue, DAPI-positive cells) B, ICES induced TUNEL staining (green, positive to TUNEL staining). Magnification: x20; scale bars: 50μm. C, Graphs demonstrating the mean ± SD of apoptotic cell density in each group (number of cells / 100 μm). *P < 0.05 versus the normal group (N), #P < 0.05, versus the scopolamine-treated group (SCOP).
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pone.0115333.g003: ICES Induced Corneal Epithelium Apoptosis.A,ICES induced caspase-3 expression in each group. (green, caspase-3–positive; blue, DAPI-positive cells) B, ICES induced TUNEL staining (green, positive to TUNEL staining). Magnification: x20; scale bars: 50μm. C, Graphs demonstrating the mean ± SD of apoptotic cell density in each group (number of cells / 100 μm). *P < 0.05 versus the normal group (N), #P < 0.05, versus the scopolamine-treated group (SCOP).

Mentions: Caspase-3 immunofluorescence and TUNEL analyses were performed to evaluate the effect of ICES exposure on corneal epithelial apoptosis. Fig. 3A shows that caspase-3 expression increased after the first 2 weeks. It peaked already at 2-weeks and remained invariant at the 4-week and 6-week time points. Similarly, TUNEL results also demonstrated that the apoptotic cells increased in the ICES group, but remained unchanged at 2-, 4- and 6-weeks. (c.f. Fig. 3B, 3C). On the other hand, corneal epithelial apoptosis in the SCOP group seemed more pronounced at all of the same time points as those in the ICES group expect for a lack of a difference after 6 weeks.


Dynamic ocular surface and lacrimal gland changes induced in experimental murine dry eye.

Xiao B, Wang Y, Reinach PS, Ren Y, Li J, Hua S, Lu H, Chen W - PLoS ONE (2015)

ICES Induced Corneal Epithelium Apoptosis.A,ICES induced caspase-3 expression in each group. (green, caspase-3–positive; blue, DAPI-positive cells) B, ICES induced TUNEL staining (green, positive to TUNEL staining). Magnification: x20; scale bars: 50μm. C, Graphs demonstrating the mean ± SD of apoptotic cell density in each group (number of cells / 100 μm). *P < 0.05 versus the normal group (N), #P < 0.05, versus the scopolamine-treated group (SCOP).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4295848&req=5

pone.0115333.g003: ICES Induced Corneal Epithelium Apoptosis.A,ICES induced caspase-3 expression in each group. (green, caspase-3–positive; blue, DAPI-positive cells) B, ICES induced TUNEL staining (green, positive to TUNEL staining). Magnification: x20; scale bars: 50μm. C, Graphs demonstrating the mean ± SD of apoptotic cell density in each group (number of cells / 100 μm). *P < 0.05 versus the normal group (N), #P < 0.05, versus the scopolamine-treated group (SCOP).
Mentions: Caspase-3 immunofluorescence and TUNEL analyses were performed to evaluate the effect of ICES exposure on corneal epithelial apoptosis. Fig. 3A shows that caspase-3 expression increased after the first 2 weeks. It peaked already at 2-weeks and remained invariant at the 4-week and 6-week time points. Similarly, TUNEL results also demonstrated that the apoptotic cells increased in the ICES group, but remained unchanged at 2-, 4- and 6-weeks. (c.f. Fig. 3B, 3C). On the other hand, corneal epithelial apoptosis in the SCOP group seemed more pronounced at all of the same time points as those in the ICES group expect for a lack of a difference after 6 weeks.

Bottom Line: To determine if there is a correlation between severity effects in these models and underlying pathophysiological responses, we compared the time dependent changes in each of these parameters that occur during a 6 week period.Interleukin-17(IL-17), IL-23, IL-6, IL-1, TNF-α, IFN-γ and TGF-β2 levels were estimated by real-time PCR measurements of conjunctival and lacrimal gland samples (LGs).Subsequently, the disease process stabilized for the next four weeks.

View Article: PubMed Central - PubMed

Affiliation: School of Ophthalmology and Optometry, Wenzhou Medical University, Zhejiang, China.

ABSTRACT
Dry eye disease can be a consequence of lacrimal gland insufficiency in Sjögren's Syndrome or increased tear film evaporation despite normal lacrimal gland function. To determine if there is a correlation between severity effects in these models and underlying pathophysiological responses, we compared the time dependent changes in each of these parameters that occur during a 6 week period. Dry eye was induced in 6-week-old female C57BL/6 mice by exposing them to an Intelligently Controlled Environmental System (ICES). Sixty mice were housed in ICES for 1, 2, 4 and 6 weeks respectively. Twelve were raised in normal environment and received subcutaneous injections of scopolamine hydrobromide (SCOP) 3 times daily for 5 days. Another sixty mice were housed in a normal environment and received no treatment. Corneal fluorescein staining along with corneal MMP-9 and caspase-3 level measurements were performed in parallel with the TUNEL assay. Interleukin-17(IL-17), IL-23, IL-6, IL-1, TNF-α, IFN-γ and TGF-β2 levels were estimated by real-time PCR measurements of conjunctival and lacrimal gland samples (LGs). Immunohistochemistry of excised LGs along with flow cytometry in cervical lymph nodes evaluated immune cell infiltration. Light and transmission electron microscopy studies evaluated LGs cytoarchitectural changes. ICES induced corneal epithelial destruction and apoptosis peaked at 2 weeks and kept stable in the following 4 weeks. In the ICES group, lacrimal gland proinflammatory cytokine level increases were much lower than those in the SCOP group. In accord with the lower proinflammatory cytokine levels, in the ICES group, lacrimal gland cytosolic vesicular density and size exceeded that in the SCOP group. ICES and SCOP induced murine dry eye effects became progressively more severe over a two week period. Subsequently, the disease process stabilized for the next four weeks. ICES induced local effects in the ocular surface, but failed to elicit lacrimal gland inflammation and cytoarchitectural changes, which accounts for less dry eye severity in the ICES model than that in the SCOP model.

Show MeSH
Related in: MedlinePlus