Expression of αA-crystallin (CRYAA) in vivo and in vitro models of age-related cataract and the effect of its silencing on HLEB3 cells

晶体蛋白 细胞凋亡 流式细胞术 转染 基因沉默 细胞生物学 活力测定 化学 自噬 分子生物学 实时聚合酶链反应 生物 细胞培养 生物化学 基因 遗传学
作者
Xiaoling Ma,Nan Yang,Can Huang,Xiangyang Li,Yifan Yang,Wenjie Jiang,Mengyi Ye,Qian Liu,Yang Niu,Ling Yuan
出处
期刊:Aging [Impact Journals, LLC]
卷期号:15 (10): 4498-4509
标识
DOI:10.18632/aging.204754
摘要

Aim: To investigate the expression of αA-crystallin (CRYAA) in age-related cataract (ARC) models and its role in lens epithelial cells (LECs).Methods: We used Flow cytometry to detect the apoptosis and cell cycle in HLEB3 cells and Real-time fluorescence quantitative polymerase chain reaction to detect the expression of CRYAA mRNA in HLEB3 and in rabbit lens. The expression of CRYAA in HLEB3 cells and rabbit lenses as well as the proteins related to apoptosis and autophagy in transfected cells were detected by western blotting. The lens structure in rabbits was investigated using hematoxylin-eosin staining. Protein thermostability assay was performed to detect the thermal stability of rabbit lens proteins. CCK- 8 assay was used to detect the viability of transfected cells, and the transfection was recorded by fluorescence photography.Results: Hydrogen peroxide can promote apoptosis and arrest the cell cycle in HLEB3 cells, and naphthalene can cause cataract formation and damage the structure of the lens in rabbits. Both ARC models can reduce the expression of CRYAA. The expression of CRYAA silencing increased apoptosis and autophagy in HLEB3 cells.
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