负性K6W泛素在镜片上皮中的主要表达是通过腺病毒载体延迟兔模型中的后囊胶质

Title:Expression of Dominant Negative K6W-Ubiquitin in the Lens Epithelium via an Adenoviral Vector Delays Posterior Capsule Opacification in a Rabbit Model

Volume: 17 Issue: 2

关键词: 后囊膜不透明（PCO），K6W-泛素，泛素 - 蛋白酶体途径（UPP），晶状体上皮细胞（LEC），腺病毒载体

摘要: Purpose: Ubiquitin is involved in cell proliferation and differentiation, and the objective of this study is to investigate the potential of dominant negative Ubiquitin (Ub) with a lysine to tryptophan mutation at the 6 position (K6W) through an adenoviral expression vector in the prevention of posterior capsule opacification (PCO) in a rabbit PCO model.

Methods: Recombinant dominant negative K6W-Ub adenovirus (RAd-K6W-Ub) with green fluorescent protein (RAd-K6W-Ub/GFP) and RAd-GFP viruses (control) were generated with QBI-HEK 293A cells. New Zealand rabbits receiving lens phacoemulsification were given an intraoperative anterior chamber injection of the viruses. The images of anterior segment photography taken by a slit lamp biomicroscopy were analyzed by posterior capsule opacification manual software (POCOman) for PCO grading. The intraocular pressure (IOP) was detected with a non-contact tonometer (NCT). The expression of α-smooth muscle actin (α-SMA) was assessed by Western blotting. Cell migration ability in cultured rabbit’s lens epithelial cells (LECs) was evaluated by scratch healing assay.

Results: The expression of GFP and Ub in the lens epithelium was markedly upregulated after 48 hours vector injection. Eyes injected with RAd-K6W-Ub showed a significantly lower PCO degree compared with controls. Meanwhile, higher IOP and corneal edema was observed in groups with a higher RAd-K6W-Ub virus dosage. The expression of α-SMA was down-regulated in the RAd-K6W-Ub eyes as compared to controls at the 15th day after injection. Cell migration was inhibited by RAd-K6W-Ub infection.

Conclusions: RAd-K6W-Ub at an appropriate dosage could inhibit the proliferation of LECs and the formation of PCO in rabbit models. However, a higher dosage of Rad- K6W-Ub viral vector caused toxic effects to the surrounding tissues, such as corneal edema and high IOP.

Cite this article as:

Expression of Dominant Negative K6W-Ubiquitin in the Lens Epithelium via an Adenoviral Vector Delays Posterior Capsule Opacification in a Rabbit Model, Current Molecular Medicine 2017; 17 (2) . https://dx.doi.org/10.2174/1566524017666170331163751

DOI https://dx.doi.org/10.2174/1566524017666170331163751	Print ISSN 1566-5240
Publisher Name Bentham Science Publisher	Online ISSN 1875-5666