Han Jung Woo, Choi Juhye, Kim Young Shin, Kim Jina, Brinkmann Ralf, Lyu Jungmook, Park Tae Kwann
Department of Ophthalmology, College of Medicine, Soonchunhyang University, Bucheon, South Korea.
Department of Medical Science, Konyang University, 158 Gwanjeodong-ro, Seo-gu, Daejeon, 320-832, Republic of Korea.
Graefes Arch Clin Exp Ophthalmol. 2018 Feb;256(2):341-353. doi: 10.1007/s00417-017-3883-7. Epub 2018 Jan 10.
This study investigated microglia and inflammatory cell responses after selective retina therapy (SRT) with microsecond-pulsed laser in comparison to continuous-wave laser photocoagulation (cwPC).
Healthy C57BL/6 J mice were treated with either a train of short pulses (SRT; 527-nm, Q-switched, 1.7-μs pulse) or a conventional thermal continuous-wave (532-nm, 100-ms pulse duration) laser. The mice were sacrificed and their eyes were enucleated 1, 3, 7, and 14 days after both laser treatments. Pattern of cell death on retinal section was evaluated by TUNEL assay, and the distribution of activated inflammatory cells and glial cells were observed under immunohistochemistry. Consecutive changes for the expression of cytokines such as IL-1β, TNF-α, and TGF-β were also examined using immunohistochemistry, and compared among each period after quantification by Western blotting.
The numbers of TUNEL-positive cells in the retinal pigment epithelium (RPE) layer did not differ in SRT and cwPC lesions, but TUNEL-positive cells in neural retinas were significantly less on SRT. Vague glial cell activation was observed in SRT-treated lesions. The population of inflammatory cells was also significantly decreased after SRT, and the cells were located in the RPE layer and subretinal space. Proinflammatory cytokines, including IL-1β and TNF-α, showed significantly lower levels after SRT; conversely, the level of TGF-β was similar to the cwPC-treated lesion.
SRT resulted in selective RPE damage without collateral thermal injury to the neural retina, and apparently produced negligible glial activation. In addition, SRT showed a markedly less inflammatory response than cwPC, which may have important therapeutic implications for several macular diseases.
本研究调查了与连续波激光光凝术(cwPC)相比,微秒脉冲激光选择性视网膜治疗(SRT)后小胶质细胞和炎症细胞的反应。
健康的C57BL/6 J小鼠接受一系列短脉冲(SRT;527纳米,调Q,1.7微秒脉冲)或传统的热连续波(532纳米,100毫秒脉冲持续时间)激光治疗。在两种激光治疗后1、3、7和14天处死小鼠并摘除眼球。通过TUNEL检测评估视网膜切片上的细胞死亡模式,并在免疫组织化学下观察活化的炎症细胞和胶质细胞的分布。还使用免疫组织化学检查细胞因子如IL-1β、TNF-α和TGF-β表达的连续变化,并在通过蛋白质印迹法定量后在每个时期之间进行比较。
视网膜色素上皮(RPE)层中TUNEL阳性细胞的数量在SRT和cwPC损伤中没有差异,但神经视网膜中的TUNEL阳性细胞在SRT中明显较少。在SRT治疗的损伤中观察到模糊的胶质细胞活化。SRT后炎症细胞的数量也显著减少,且细胞位于RPE层和视网膜下间隙。包括IL-1β和TNF-α在内的促炎细胞因子在SRT后水平显著降低;相反,TGF-β的水平与cwPC治疗的损伤相似。
SRT导致选择性RPE损伤而不对神经视网膜造成附带热损伤,并且显然产生可忽略不计的胶质细胞活化。此外,SRT显示出比cwPC明显更少的炎症反应,这可能对几种黄斑疾病具有重要的治疗意义。