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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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Flow cytometry analysis of cervical lymph nodes.Flow cytometry analysis of freshly isolated cells from cervical lymph nodes stained with CD4-PEand CD8-FITC conjugated antibody in each group. Numbers in the quadrants indicate the percentage of cells. Similar percentages of CD4+lymphocytes were observed in the cervical lymph nodes in all groups. Percentages of CD8+lymphocytes in ICES 2-week, 4-week and 6-week groups were also at baseline level, while that in the SCOP group was lower than the normal level.
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pone.0115333.g008: Flow cytometry analysis of cervical lymph nodes.Flow cytometry analysis of freshly isolated cells from cervical lymph nodes stained with CD4-PEand CD8-FITC conjugated antibody in each group. Numbers in the quadrants indicate the percentage of cells. Similar percentages of CD4+lymphocytes were observed in the cervical lymph nodes in all groups. Percentages of CD8+lymphocytes in ICES 2-week, 4-week and 6-week groups were also at baseline level, while that in the SCOP group was lower than the normal level.

Mentions: Flow cytometry analysis of CLN cells stained for CD4 and CD8 was performed. Similar percentages of CD4+ lymphocytes were observed in these cells in the N, ICES and SCOP groups at all the different times. CD8+ lymphocyte percentages in the ICES group at 2-, 4- and 6-weeks were in all cases invariant at its baseline level, while in the SCOP group its level was even lower than in the N group (Fig. 8). 


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)

Flow cytometry analysis of cervical lymph nodes.Flow cytometry analysis of freshly isolated cells from cervical lymph nodes stained with CD4-PEand CD8-FITC conjugated antibody in each group. Numbers in the quadrants indicate the percentage of cells. Similar percentages of CD4+lymphocytes were observed in the cervical lymph nodes in all groups. Percentages of CD8+lymphocytes in ICES 2-week, 4-week and 6-week groups were also at baseline level, while that in the SCOP group was lower than the normal level.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0115333.g008: Flow cytometry analysis of cervical lymph nodes.Flow cytometry analysis of freshly isolated cells from cervical lymph nodes stained with CD4-PEand CD8-FITC conjugated antibody in each group. Numbers in the quadrants indicate the percentage of cells. Similar percentages of CD4+lymphocytes were observed in the cervical lymph nodes in all groups. Percentages of CD8+lymphocytes in ICES 2-week, 4-week and 6-week groups were also at baseline level, while that in the SCOP group was lower than the normal level.
Mentions: Flow cytometry analysis of CLN cells stained for CD4 and CD8 was performed. Similar percentages of CD4+ lymphocytes were observed in these cells in the N, ICES and SCOP groups at all the different times. CD8+ lymphocyte percentages in the ICES group at 2-, 4- and 6-weeks were in all cases invariant at its baseline level, while in the SCOP group its level was even lower than in the N group (Fig. 8). 

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