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兔双侧主泪腺切除术后的泪液分泌

Tear Production After Bilateral Main Lacrimal Gland Resection in Rabbits.

作者信息

Bhattacharya Dhruva, Ning Yuan, Zhao Fangkun, Stevenson William, Chen Rongji, Zhang Jinsong, Wang Mingwu

机构信息

Department of Ophthalmology and Vision Science University of Arizona College of Medicine, Tucson, Arizona, United States.

Department of Ophthalmology and Vision Science University of Arizona College of Medicine, Tucson, Arizona, United States 2Department of Ophthalmology, the Fourth Affiliated Hospital of China Medical University, Eye Hospital of China Medical University, Ke.

出版信息

Invest Ophthalmol Vis Sci. 2015 Dec;56(13):7774-83. doi: 10.1167/iovs.15-17550.

Abstract

PURPOSE

This study reports tear compensation observed in rabbits with bilateral resection of main lacrimal gland (LG) and explored the potential mechanisms.

METHODS

Dry eye conditions were created by resection of nictitating membrane (NM), Harderian gland (HG), and main LG in eight (16 eyes) male New Zealand White rabbits. In addition to Schirmer test, Periodic acid-Schiff (PAS) staining, and ocular surface staining with fluorescein and rose Bengal, conjunctival impression cytology was employed before and up to 4 months after excision (AE). Using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), expression of inflammatory biomarkers (IL-1β, TNF-α, and matrix metalloproteinase-9) were monitored. Further, involvement of ionic and water transporters were investigated in conjunctival epithelium.

RESULTS

Significant dry eye phenotypes in rabbits were observed 1 month AE, which corroborated with elevated biomarker mRNA expression. However, Schirmer test score and goblet cell numbers never decreased AE in conjunctival epithelium. Moreover, ocular surface staining, and biomarker expression declined to baseline in over 4 months AE. No upregulation was observed of the following conjunctival ionic transporters: cystic fibrosis transmembrane conductance regulator, sodium potassium chloride cotransporters, sodium potassium ATPase, and epithelial sodium channels. Instead, aquaporin (AQP) 4 and AQP5 were upregulated. Immunolocalization and immune blotting of AQP4 was demonstrated in rabbit conjunctival epithelium.

CONCLUSIONS

In the absence of NM, HG, and main LG in rabbits, tear secretion was not decreased and significant improvement of dry eye phenotypes observed with time AE. Conjunctival AQPs are possibly involved in a compensatory tear fluid production.

摘要

目的

本研究报告了双侧切除主泪腺(LG)的兔子中观察到的泪液代偿情况,并探讨了其潜在机制。

方法

对8只(16只眼)雄性新西兰白兔切除瞬膜(NM)、哈德氏腺(HG)和主泪腺,以建立干眼状态。除了进行泪液分泌试验、过碘酸-希夫(PAS)染色以及用荧光素和孟加拉玫瑰红进行眼表染色外,在切除前(AE)及切除后长达4个月进行结膜印迹细胞学检查。使用逆转录-定量聚合酶链反应(RT-qPCR)监测炎症生物标志物(IL-1β、TNF-α和基质金属蛋白酶-9)的表达。此外,研究结膜上皮中离子和水转运蛋白的参与情况。

结果

切除后1个月观察到兔子出现明显的干眼表型,这与生物标志物mRNA表达升高相一致。然而,泪液分泌试验评分和结膜上皮杯状细胞数量在切除后从未减少。此外,眼表染色和生物标志物表达在切除后4个月以上降至基线。未观察到以下结膜离子转运蛋白的上调:囊性纤维化跨膜传导调节因子、钠钾氯共转运蛋白、钠钾ATP酶和上皮钠通道。相反,水通道蛋白(AQP)4和AQP5上调。在兔结膜上皮中证实了AQP4的免疫定位和免疫印迹。

结论

在兔子缺失瞬膜、哈德氏腺和主泪腺的情况下,泪液分泌未减少,且随着切除后时间的推移干眼表型有显著改善。结膜水通道蛋白可能参与泪液的代偿性产生。

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