变色人工晶状体对可见光诱导人视网膜色素上皮细胞损伤的保护作用

摘要

目的 探讨变色人工晶状体(IOL)对可见光诱导人视网膜色素上皮(RPE)细胞损伤的保护作用.方法 实验研究.体外培养人RPE细胞(ARPE-19),用同一光照度的可见光照射,在光通路上置不同IOL.RPE细胞分为5组:阴性对照组(无光照,无IOL)、阳性对照组(照光,无IOL)、无色IOL组(照光,FV-60A IOL)、变色IOL组(照光,BV-60A IOL)、黄色IOL组(照光,Acrysof Natural IOL).用MTT法检测RPE细胞活力,透射电子显微镜、脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)检测RPE细胞凋亡情况；Hoechst染色计算凋亡指数(AI)；酶联免疫吸附法检测血管内皮生长因子(VEGF)、色素上皮衍生因子(PEDF)的浓度.对数据进行方差分析,多个均数间两两比较用q检验.结果 阴性对照组、阳性对照组、无色IOL组、变色IOL组、黄色IOL组细胞吸光度(A)值分别为0.61 ±0.01、0.39 ±0.01、0.46±0.01、0.57 ±0.01及0.58±0.02; TUNEL染色和透射电镜下检查均发现光照后细胞出现凋亡改变.5个组细胞的AI值分别是(0.99±0.65)％、(9.28±0.59)％、(6.54±0.37)％、(4.69±0.74)％及(2.72±0.67)％.5个组VEGF浓度分别为(3931.64±25.26) ng/L、(5459.38±42.69) ng/L、(4800.55 ±42.12) ng/L、(4561.21 ±46.27) ng/L及(4420.35±28.66) ng/L；PEDF浓度分别为(182.85±1.68) μg/L、(156.52 ±0.64) μg/L、(155.57±3.43)μg/L、(167.57 ±2.88) μg/L及(170.90 ±2.99) μg/L.变色IOL组与黄色IOL组比较,细胞活力(q=0.94)、PEDF浓度(q=1.51)的差异无统计学意义(P＞0.05)；AI值(q=5.04)、VEGF浓度(q=6.78)的差异有统计学意义(P＜0.05)；变色IOL组与无色IOL组比较,细胞活力(q=18.92)、AI值(q=4.78)、VEGF浓度(q=11.52)及PEDF浓度(q=4.96)的差异有统计学意义(P＜0.05).结论 变色IOL能减轻RPE细胞的急性光损伤程度,保护作用优于紫外线阻断型IOL,弱于蓝光滤过型IOL.%Objective To evaluate the protective effect of photochromic intraocular lens (IOL) on visible light-induced lesion in cultured retinal pigment epithelium (RPE).Methods It was a experimental study.A human retinal pigment epithelium cell line (ARPE-19) was used to study the protective role.Cells from the same passage were divided into the control,light exposure only,light exposure plus colorless IOL (FV-60A),light exposure plus photochromic IOL (BV-60A),light exposure plus yellow IOL (Acrysof Natural IOL) groups.The treatments were performed when the cells grew to 90％ confluency.All cells were irradiated with visible light at an illumination intensity of 60000-61000 lux.After the treatments,we assessed 1:cells' viability by methyl thiazolyl tetrazolium (MTT) assay; 2:the extent of apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL),observing changes in the ultrastructure by transmission electron microscopy (TEM),and performing Hoechst staining to measure the apoptosis index (AI) ; 3:examining vascular endothelial growth factor (VEGF) and pigment epitheliumderived factor (PEDF) levels in the culture medium by enzyme linked immunosorbent assay (ELISA).Results Measurements of the cell viability of the five groups were (0.61 ± 0.01),(0.39 ± 0.01),(0.46 ±0.01),(0.57 ±0.01),and (0.58 ±0.02),respectively.Apoptosis was significantly activated by visible light as observed in TUNEL staining and TEM.The AIs of the five groups were (0.99 ± 0.65) ％,(9.28 ± 0.59) ％,(6.54 ± 0.37) ％,(4.69 ± 0.74) ％,and (2.72 ± 0.67) ％,respectively.The levels of VEGF in the five groups were (3931.64 ± 25.26) ng/L,(5459.38 ± 42.69) ng/L,(4800.55 ±42.12) ng/L,(4561.21 ±46.27) ng/L,and (4420.35 ±28.66) ng/L,respectively.The levels of PEDF in the five groups were (182.85 ± 1.68) μg/L,(156.52 ±0.64) μg/L,(155.57 ±3.43) μg/L,(167.57 ±2.88) μg/L,and (170.90 ±2.99) μg/L,respectively.Comparing the photochromic IOL group with the yellow IOL group,there was no difference in cell viability (q =0.94) and PEDF levels (q =1.51)(P ＞ 0.05),but there was a significant difference between the AI (q =5.04) and VEGF levels (q =6.78)(P ＜ 0.05).Comparing the photochromic IOL group with the colorless IOL group,there were significant differences in cell viability (q =18.92),AI(q =4.78),VEGF (q =11.52) and PEDF levels (q =4.96)(P ＜ 0.05).Conclusions Photochromic IOL can abate the degree of acute photodamage of RPE.The protective efficiency was the highest when using the yellow Acrysof Natural IOL,followed by photochromic IOL and colorless IOL.(Chin J Ophthalmol,2013,49:410-415)