Inagaki Keiji, Shuo Takuya, Katakura Kanae, Ebihara Nobuyuki, Murakami Akira, Ohkoshi Kishiko
Department of Ophthalmology, St. Luke's International Hospital, 9-1 Akashi-cho, Chuo-ku, Tokyo 104-8560, Japan ; Department of Ophthalmology, Juntendo University Graduate School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421, Japan.
Institute for Medical Innovation, St. Luke's International University, 10-1 Akashi-cho, Chuo-ku, Tokyo 104-8560, Japan.
J Ophthalmol. 2015;2015:729792. doi: 10.1155/2015/729792. Epub 2015 Nov 30.
Purpose/Aim of the Study. Subthreshold micropulse diode laser photocoagulation is an effective treatment for macular edema. The molecular mechanisms underlying treatment success are poorly understood. Therefore, we investigated the effects of sublethal laser energy doses on a single layer of densely cultured ARPE-19 cells as a model of the human retinal pigment epithelium (RPE). Materials and Methods. A single layer of densely cultured human ARPE-19 cells was perpendicularly irradiated with a micropulse diode laser. Nonirradiated cells served as controls. Sublethal laser energy was applied to form a photocoagulation-like area in the cultured cell layers. Hsp70 expression was evaluated using quantitative polymerase chain reaction and immunocytochemistry. Results. Photocoagulation-like areas were successfully created in cultured ARPE-19 cell layers using sublethal laser energy with our laser irradiation system. Hsp70 mRNA expression in cell layers was induced within 30 min of laser irradiation, peaking at 3 h after irradiation. This increase was dependent on the number of laser pulses. Hsp70 upregulation was not observed in untreated cell layers. Immunostaining indicated that Hsp70 expression occurred concentrically around laser irradiation sites and persisted for 24 h following irradiation. Conclusion. Sublethal photothermal stimulation with a micropulse laser may facilitate Hsp70 expression in the RPE without inducing cellular damage.
研究目的。阈下微脉冲二极管激光光凝术是治疗黄斑水肿的一种有效方法。治疗成功背后的分子机制尚不清楚。因此，我们以人视网膜色素上皮（RPE）模型——单层密集培养的ARPE - 19细胞为研究对象，探讨亚致死激光能量剂量的影响。材料与方法。用微脉冲二极管激光垂直照射单层密集培养的人ARPE - 19细胞。未照射的细胞作为对照。施加亚致死激光能量在培养的细胞层中形成类似光凝的区域。使用定量聚合酶链反应和免疫细胞化学评估热休克蛋白70（Hsp70）的表达。结果。使用我们的激光照射系统，通过亚致死激光能量在培养的ARPE - 19细胞层中成功创建了类似光凝的区域。激光照射后30分钟内，细胞层中Hsp70 mRNA表达开始诱导，照射后3小时达到峰值。这种增加依赖于激光脉冲的数量。在未处理的细胞层中未观察到Hsp70上调。免疫染色表明，Hsp70表达在激光照射部位周围呈同心圆状出现，并在照射后持续24小时。结论。微脉冲激光的亚致死光热刺激可能促进RPE中Hsp70的表达而不诱导细胞损伤。
