论文部分内容阅读
目的 研究可见光照对培养的人视网膜色素上皮 (retinal pigment epithelium,RPE)细胞凋亡的影响。 方法 以白色荧光灯为光源 ,用 5 0 0 lx、(2 0 0 0± 5 0 0 ) lx及 (340 0± 2 0 0 ) lx不同光照强度 ,按不同的光照时间 (无光照、6、12、2 4 h)照射培养的人 RPE细胞。利用终末脱氧核糖核苷酸转移酶介导的原位缺口末端标记 (terminal deoxynucleotidyl transferase mediated d UTP nick end labelling,TUNEL)、荧光素标记的连接素 V/碘化丙锭 (Annexin V- fluorescein isothiocyanate/ Propidium iodium,AnnexinV- FITC/ PI)双染色流式细胞测定、相差倒置显微镜等手段观察 RPE细胞凋亡 (分为光照后 6、12、2 4、36 h组 )。 结果 可观察到 RPE细胞出现两种死亡形式 ,凋亡与坏死。 (1)低于一定阈值 (5 0 0 lx)的光照对细胞损伤较轻 ,细胞凋亡及坏死随光照强度的增加而增加。 (2 )在较短的光照时间 (6 h和 12 h)内 ,细胞死亡的增加以凋亡为主 ;随着光照时间的延长 ,细胞坏死逐渐明显。 (3)随着光照后培养时间的延长 ,细胞凋亡明显增加 (P<0 .0 5 )。光照后 6、12、2 4 h的损伤改变以凋亡为主 ,但随时间的延长 ,凋亡继发性坏死的增加显著。光照后 36 h,细胞的坏死数显著增高 (P<0 .0 1)。 结论 可见?
Objective To study the effect of visible light on the apoptosis of cultured human retinal pigment epithelium (RPE) cells. Methods The white fluorescent lamp was used as a light source, with different light intensity of 500 lx, (20000 ± 5000) lx and (34000 ± 2000) lx, with different illumination time , 24 h) cultured human RPE cells. Using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL), the binding of fluorescein-labeled linker V / propidium iodide (Annexin V-fluorescein isothiocyanate / Propidium iodium, AnnexinV-FITC / PI) were used to observe the apoptosis of RPE cells (divided into 6, 12, 24, 36 h groups after light exposure) by means of inverted phase contrast microscope. Results Two forms of death, apoptosis and necrosis were observed in RPE cells. (1) Light below a certain threshold (500 lx) had less damage to cells, and apoptosis and necrosis increased with light intensity. (2) The apoptosis was dominated by the increase of cell death in shorter illumination time (6 h and 12 h). As the illumination time prolonged, cell necrosis became obvious. (3) With the prolongation of culture time after light irradiation, apoptosis increased significantly (P <0.05). At 6, 12 and 24 h after light irradiation, the change of apoptosis was mainly apoptosis, but with the prolongation of time, the apoptosis necrosis increased significantly. After 36 h of illumination, the number of necrotic cells was significantly increased (P <0.01). Can be seen?