Jang Sun Young, Cho In Hwan, Yang Jin Young, Park Ha Yan, Woo Sang Earn, Madrakhimov Sanjar Batirovich, Chang Hun Soo, Lyu Jungmook, Park Tae Kwann
Department of Ophthalmology, Soonchunhyang University Hospital Bucheon, Soonchunhyang University College of Medicine, #170 Jomaru-ro, Bucheon-si, Gyeonggi-do, 14584, South Korea.
Department of Ophthalmology, College of Medicine, Soonchunhyang University, Cheonan-si, Chungcheongnam-do, South Korea.
Lasers Med Sci. 2019 Feb;34(1):179-190. doi: 10.1007/s10103-018-2680-9. Epub 2018 Nov 29.
To investigate the characteristics of regenerated retinal pigment epithelial (RPE) cells after retinal laser photocoagulation in diabetic mice. C57BL/6J mice were used to induce diabetes using intraperitoneal injection of streptozotocin. The proliferation of RPE cells after laser photocoagulation was determined using the 5-ethynyl-2'-deoxyuridine (EdU) assay in both diabetic and wild-type mice. The morphological changes of RPE cells were evaluated by using Voronoi diagram from immunostaining for ß-catenin. Characteristics of regenerated cells were evaluated by quantifying the mRNA and protein levels of RPE and epithelial-mesenchymal transition (EMT) markers. There were significantly less EdU-positive cells in laser-treated areas in diabetic mice than wild-type mice. Hexagonality was extensively lost in diabetic mice. Many EdU-positive cells were co-localized with Otx2-positive cells in the center of the laser-treated areas in wild-type mice, but only EdU-positive cells were widely distributed in diabetic mice. Quantitative analysis of mRNA and protein levels showed that the expression levels of RPE markers, Pax6, Mitf, and Otx2, were significantly decreased in RPE of diabetic mice compared with that of wild-type mice, whereas the expression levels of EMT markers, vimentin and fibronectin, were significantly increased. The proliferation and hexagonality of regenerating RPE cells were impaired after laser photocoagulation, and the regenerated RPE cells lost their original properties in diabetic mice. Further clinical research is needed to elucidate the RPE response after laser photocoagulation in diabetic patients.
研究糖尿病小鼠视网膜激光光凝后再生视网膜色素上皮（RPE）细胞的特征。采用C57BL/6J小鼠，通过腹腔注射链脲佐菌素诱导糖尿病。使用5-乙炔基-2'-脱氧尿苷（EdU）检测法测定糖尿病小鼠和野生型小鼠激光光凝后RPE细胞的增殖情况。通过对β-连环蛋白免疫染色的Voronoi图评估RPE细胞的形态变化。通过定量RPE和上皮-间质转化（EMT）标志物的mRNA和蛋白质水平来评估再生细胞的特征。糖尿病小鼠激光治疗区域的EdU阳性细胞明显少于野生型小鼠。糖尿病小鼠的六边形结构大量丧失。在野生型小鼠激光治疗区域的中心，许多EdU阳性细胞与Otx2阳性细胞共定位，但在糖尿病小鼠中只有EdU阳性细胞广泛分布。mRNA和蛋白质水平的定量分析表明，与野生型小鼠相比，糖尿病小鼠RPE中RPE标志物Pax6、Mitf和Otx2的表达水平显著降低，而EMT标志物波形蛋白和纤连蛋白的表达水平显著升高。激光光凝后再生RPE细胞的增殖和六边形结构受损，且糖尿病小鼠中再生的RPE细胞失去了其原始特性。需要进一步的临床研究来阐明糖尿病患者激光光凝后的RPE反应。
