MiRNA-155-5p Reduces Corneal Epithelial Permeability by Remodeling Epithelial Tight Junctions during Corneal Wound Healing

被引:21
|
作者
Wang, Feng [1 ,2 ,3 ]
Wang, Duomei [2 ,3 ]
Song, Meng [2 ,3 ]
Zhou, Qing [2 ,3 ]
Liao, Rongfeng [1 ]
Wang, Yuan [2 ,3 ]
机构
[1] Anhui Med Univ, Dept Ophthalmol, Affiliated Hosp 1, Hefei 230022, Anhui, Peoples R China
[2] Anhui Med Univ, Lab Mol Biol, Hefei 230032, Anhui, Peoples R China
[3] Anhui Med Univ, Dept Biochem, Hefei 230032, Anhui, Peoples R China
基金
中国国家自然科学基金;
关键词
Cornea; epithelium; wound healing; miR-155-5p; tight junction; myosin light chain kinase; LIGHT-CHAIN KINASE; ACTIN; PROLIFERATION; MICRORNA-155; MIGRATION; PATHWAY;
D O I
10.1080/02713683.2019.1707229
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
Purpose: Corneal epithelial cells play a vital role in the function of the cornea by forming a physical barrier to protect the eye from invasion by external pathogenic agents. A recent study showed that miR-155 promotes cutaneous wound healing. However, its function in corneal epithelial wound healing is unknown. The present study examined whether miR-155-5p reduces corneal epithelial permeability by remodeling epithelial tight junctions during corneal wound healing. Materials and Methods: Rat corneal wounds were produced by removing the central corneal epithelium with a blunt scalpel blade under a dissecting microscope. One eye of each rat was treated with topical miR-155-5p, and the other eye was treated with topical agomir negative control for 3 days before and after corneal epithelial wounding. Corneal epithelial permeability was assessed by the macromolecular osmosis method. Expression of zona occludens 1 (ZO-1), occludin, and myosin light chain kinase (MLCK) and phosphorylation of myosin light chain (MLC) were detected by Western blot. Human corneal epithelial (HCE) cells were cultured in the upper chamber of Transwell filters, and transepithelial electrical resistance (TER) was measured using a voltohmmeter. The distribution of ZO-1 and occludin in HCE cells treated with miR-155-5p was determined by immunofluorescence. Results: miR-155-5p significantly promoted the repair of corneal epithelial injury and reduced the permeability of the corneal epithelium. It significantly decreased expression of MLCK and phosphorylation of MLC and increased expression of the tight junction proteins ZO-1 and occludin in corneal epithelial cells during corneal wound healing. miR-155-5p significantly increased TER, decreased MLCK expression and MLC phosphorylation, increased ZO-1 and occludin expression, and promoted anchoring of tight junction proteins in the cell membrane and remodeling in HEC cells. Conclusions: Our results suggest that miR-155-5p reduced corneal permeability and accelerated the recovery of corneal epithelial wounds by decreasing the expression of MLCK and phosphorylation of MLC and by remodeling tight junctions.
引用
收藏
页码:904 / 913
页数:10
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