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外源性αB晶状体蛋白对小梁网细胞结构和功能的影响

Effect of Exogenous Alpha-B Crystallin on the Structures and Functions of Trabecular Meshwork Cells.

作者信息

Xu Hui, Zhu Li, Wang Yu, Bao Yongzhen

机构信息

Department of Ophthalmology, Peking University People's Hospital, Eye diseases and Optometry Institute, Beijing Key Laboratory of Diagnosis and Therapy of Retina and Choroid Diseases, College of Optometry, Peking University Health Science Center, Beijing, China.

出版信息

J Ophthalmol. 2018 Apr 17;2018:7875318. doi: 10.1155/2018/7875318. eCollection 2018.

Abstract

PURPOSE

Secondary open-angle glaucoma may develop as a postoperative complication of early childhood cataract surgery. Its mechanism is poorly understood. Surgical removal of cataracts is typically incomplete, and we estimate that this disease is associated with alpha-B crystallin (CRYAB) secreted from the retained lens material. This study, for the first time, focused on the role of CRYAB in undesired changes of the structures and functions in trabecular meshwork (TM) cells.

METHODS

Cell proliferation and migration were assessed using a cell counting kit-8 (CCK8) and transwell assay analysis, respectively. Immunofluorescence (IF), quantitative real-time PCR (Rt-qPCR), and Western blot were performed to determine the effect of CRYAB on F-actin, tight junctions, and the expression of epithelial to mesenchymal transition- (EMT-) associated proteins in TM cells.

RESULTS

CRYAB promoted proliferation ( < 0.0001), migration ( < 0.001), and F-actin reorganization in TM cells. There were statistically significant increases in the mRNA and protein levels of zo-1, cadherin-N, and vimentin (all < 0.0001) and cadherin-E decreased ( < 0.0001) and the mRNA level of claudin-1 increased ( < 0.0001) compared to those of the control group.

CONCLUSION

All of the changes in structures and functions first observed in the TM cells after exposure to CRYAB resembled alterations seen in primary open-angle glaucoma, suggesting that CRYAB might be related to the pathogenesis of secondary open-angle glaucoma after congenital cataract surgery.

摘要

目的

继发性开角型青光眼可能作为儿童早期白内障手术的术后并发症出现。其发病机制尚不清楚。白内障手术通常不能完全清除晶状体,我们估计这种疾病与残留晶状体物质分泌的α-B晶状体蛋白(CRYAB)有关。本研究首次聚焦于CRYAB在小梁网(TM)细胞结构和功能不良变化中的作用。

方法

分别使用细胞计数试剂盒-8(CCK8)和Transwell分析评估细胞增殖和迁移。进行免疫荧光(IF)、定量实时PCR(Rt-qPCR)和蛋白质印迹法,以确定CRYAB对TM细胞中F-肌动蛋白、紧密连接以及上皮-间质转化(EMT)相关蛋白表达的影响。

结果

CRYAB促进TM细胞的增殖(<0.0001)、迁移(<0.001)和F-肌动蛋白重组。与对照组相比,zo-1、钙黏蛋白-N和波形蛋白的mRNA和蛋白水平均有统计学显著升高(均<0.0001),钙黏蛋白-E降低(<0.0001),紧密连接蛋白-1的mRNA水平升高(<0.0001)。

结论

暴露于CRYAB后首次在TM细胞中观察到的所有结构和功能变化类似于原发性开角型青光眼中所见的改变,这表明CRYAB可能与先天性白内障手术后继发性开角型青光眼的发病机制有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7aa8/5932433/daee7d36aea6/JOPH2018-7875318.001.jpg

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