Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Departments of Ophthalmology and Pathology, New York Eye and Ear Infirmary of Mount Sinai, New York, NY, USA.
Biomed Res Int. 2017;2017:2647879. doi: 10.1155/2017/2647879. Epub 2017 May 29.
To identify an effective method to prevent myopia progression by characterizing the regulation of matrix metalloproteinase- (MMP-) 2 expression and its secretion from scleral fibroblasts and retinal pigment epithelium (RPE) cells by miR-29a.
The effects of miR-29a on the growth of scleral fibroblasts and RPE cells were assessed using the cell counting kit-8. The changes in MMP-2 mRNA levels in scleral fibroblasts and RPE cells after transfection with miR-29a mimics or inhibitor were measured by quantitative PCR. Enzyme-linked immunosorbent assays were used to determine the changes in MMP-2 secretion from scleral fibroblasts and RPE cells after transfection with miR-29a mimics or inhibitor.
The miR-29a mimics or inhibitor did not significantly alter the growth of scleral fibroblasts or RPE cells at 24, 48, or 72 hours after transfection. MMP-2 mRNA levels were significantly decreased in scleral fibroblasts and RPE cells transfected with the miR-29a mimics. The secretion of MMP-2 by scleral fibroblasts and RPE cells was significantly decreased in cells transfected with the miR-29a mimics.
Suppression of scleral fibroblast and RPE cell expression and secretion of MMP-2 by miR-29a can be used as a therapeutic target for the prevention and treatment of myopia.
通过鉴定 miR-29a 对基质金属蛋白酶-2(MMP-2)表达及其在巩膜成纤维细胞和视网膜色素上皮(RPE)细胞中分泌的调控作用,找到一种有效预防近视进展的方法。
通过细胞计数试剂盒-8 评估 miR-29a 对巩膜成纤维细胞和 RPE 细胞生长的影响。用定量 PCR 检测转染 miR-29a 模拟物或抑制剂后巩膜成纤维细胞和 RPE 细胞中 MMP-2 mRNA 水平的变化。用酶联免疫吸附试验检测转染 miR-29a 模拟物或抑制剂后巩膜成纤维细胞和 RPE 细胞中 MMP-2 分泌的变化。
转染 miR-29a 模拟物或抑制剂 24、48 或 72 小时后,对巩膜成纤维细胞或 RPE 细胞的生长没有明显影响。转染 miR-29a 模拟物后,巩膜成纤维细胞和 RPE 细胞中 MMP-2 mRNA 水平显著降低。转染 miR-29a 模拟物后,巩膜成纤维细胞和 RPE 细胞中 MMP-2 的分泌明显减少。
miR-29a 抑制巩膜成纤维细胞和 RPE 细胞 MMP-2 的表达和分泌,可作为预防和治疗近视的治疗靶点。