Biological Modulation of Mouse RPE Cells in Response to Subthreshold Diode Micropulse Laser Treatment
Biological Modulation of Mouse RPE Cells in Response to Subthreshold Diode Micropulse Laser Treatment
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小鼠 RPE 细胞响应亚阈值二极管微脉冲激光治疗的生物调节
DOI：
10.1007/s12013-015-0675-8
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发表时间：
2015-11-01
影响因子：
2.6
通讯作者：
Ding, Qin
中科院分区：
文献类型：
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作者：
Li, Zhouyue;Song, Yanping;Ding, Qin
许多临床试验已经证明了阈值下光疗在视网膜血管疾病（如糖尿病视网膜病变）中的有效性，并且没有可见的损伤。目的：研究阈下半导体微脉冲激光（SDM）对小鼠视网膜色素上皮（RPE）细胞的作用。还探索了响应于SDM的血管生成调节细胞因子的表达。通过用MTT测定法测量细胞活力来选择毒性最小的激光剂量，并选择5%占空比（DC）用于进一步的实验。用0 ~ 400 mW的激光诱导辐射处理RPE细胞24 h。流式细胞仪检测RPE细胞凋亡率。Western Blotting和real-time PCR分别检测血管内皮生长因子A（VEGF-A）、转化生长因子β（TGF-β）、碱性成纤维细胞生长因子（bFGF）和色素上皮衍生因子（PEDF）的表达。激光照射24 h后，细胞活力呈剂量依赖性降低，与对照组相比差异有显著性（P< 0.05）。此外，与对照组相比，100和200 mW激光和5% DC处理对细胞活力和凋亡无显著影响（P> 0.05）。血管生成刺激因子（VEGF-A、TGF-β和bFGF）的蛋白和mRNA表达显著下调（P< 0.05），而血管生成抑制因子（PEDF）的蛋白和mRNA表达显著上调（P< 0.05）。不同强度的激光照射后细胞凋亡率差异无统计学意义（P> 0.05）。我们的研究结果表明，SDM治疗的RPE细胞抑制脉络膜新生血管促进细胞因子的表达和上调血管生成抑制剂，PEDF，而不损害细胞。需要进一步的研究来理解其机制并优化SDM作为视网膜血管疾病治疗的新方法的使用。
Many clinical trials have demonstrated the effectiveness of subthreshold phototherapy with no visible damage in retinal vascular diseases, such as diabetic retinopathy. We aimed primarily to investigate the effect of subthreshold diode micropulse laser (SDM) treatment on mouse retinal pigmented epithelium (RPE) cells. The expression of angiogenesis-modulating cytokines in response to SDM was also explored. The least toxic laser dose was selected by measuring cell viability with MTT assay and 5 % duty cycle (DC) was chosen for use in further experiments. RPE cells were treated with laser-induced radiation ranging from 0 to 400 mW for 24 h. The apoptotic rate of RPE cells was assessed by flow cytometry. Expressions of vascular endothelial growth factor A (VEGF-A), transforming growth factor beta (TGF-β), basic fibroblast growth factor (bFGF), and pigment epithelium-derived factor (PEDF) were determined by Western Blotting and real-time PCR, respectively. After 24 h of laser irradiation, cell viability was reduced dose dependently and the effect was significant compared to the controls (P< 0.05). In addition, laser treatment with intensities of 100 and 200 mW with DC of 5 % produced no significant effect on cell viability and apoptosis as compared with the control group (P> 0.05). The protein and mRNA expressions of angiogenic stimulators (VEGF-A, TGF-β, and bFGF) were significantly down-regulated (P< 0.05), whereas those of the angiogenic inhibitor (PEDF) were up-regulated (P< 0.05). No significant difference was found between the cells treated with different intensities of laser radiation (P> 0.05). Our results showed that SDM treatment of the RPE cells suppressed the expression of choroid neovasculization-promoting cytokines and up-regulated the angiogenic inhibitor, PEDF without damaging the cells. Further investigation is needed to understand the mechanism and to optimize the use of SDM as a novel method of treatment for retinal vascular diseases.