多聚赖氨酸-乙二胺四乙酸交联物抑制兔晶状体上皮细胞生长的研究

摘要

背景 后囊膜混浊(PCO)是现代白内障囊外摘出术后引起视力下降的主要原因。研究证实多聚赖氨酸-乙二胺四乙酸( EDTA)交联物(PLE)能够降低兔PCO的发生率。 目的 研究PLE对体外培养的兔晶状体上皮细胞(LECs)生长的抑制作用及有效药物浓度。 方法 取3月龄新西兰白兔晶状体前囊膜进行体外植块培养获得兔LECs并进行传代，取第2～3代传代培养的LECs消化后按1×105个/ml密度接种于96孔培养板中。在培养皿中加入12.5、25.0、50.0、100.0 μmol/L的PLE，培养皿中加入DMSO培养液作为对照组。PLE作用48 h用MTT比色法测定PLE对兔LECs增生的抑制作用并计算各浓度PLE组的吸光度(A490)值及其抑制率。结果 ≤25.0 μmol/L PLE组可见细胞生长及形态改变不明显。≥50.0μmol/L PLE各组可见细胞生长及形态有明显改变，增生缓慢，生长差，数量减少，贴壁能力减弱。12.5、25.0、50.0、100.0 μmol/L的PLE组LECs的A490值分别为0.278±0.013、0.266±0.028、0.260±0.022和0.247±0.012，均明显低于DMSO对照组的0.311±0.038(P=0.035、0.011、0.009、0.013)，且呈明显的剂量-效应依赖关系。12.5、25.0、50.0、100.0 μmol/L PLE组对兔LECs的抑制率分别为10.61％、14.47％、16.40％和20.58％。结论PLE可以浓度依赖的方式抑制兔LECs的生长，可为临床筛选出防治PCO的药物提供依据。%Background Posterior capsule opacification(PCO) is the main cause inducing low vision after extacapsular cataract extraction. Our previous study determined that polylysine-ethylene diamine tetraacetic acid (EDTA) (PLE) can suppress the incidence of PCO. Objective The goal of this experiment was to investigate the inhibition of polylysine-EDTA on rabbit lens epithelial cells (LECs) proliferation in vitro and the effective concentrations of polylysine-EDTA. Methods The anterior capsular membranes from 10 3-month-old clean New Zealand white rabbits were digested and then cultured to obtain the LECs. The second and third generation of LECs were inoculated on the 96-hole culture plate with the cell density of the 1 × 105/ml. 12.5,25.0,50. 0,100. 0 μmol/Lof PLE were added into the culture medium for 48 hours respectively,and the DMSO medium was used at the same way as the control group. The proliferation of the LECs was then detected by MTT method and the inhibitory rate of PLE on LECs growth was calculated. Results LECs grew at a near normal state in ≤25.0 μmol/L PLE groups,however,cultured LECs were out of shape and the numbers decreased with the weakened adhesion ability in ≥50.0 μ mol/L PLE groups. The A490 values of LECs were 0. 278±0. 013,0. 266±0. 028,0. 260±0. 022 and 0. 247±0. 012 in 12. 5,25.0,50. 0, 100. 0 μmol/L polylysine-EDTA groups respectively and were lower than 0. 311 ±0. 038 of DMSO control group( P=0. 035,0. 011,0. 009,0.013 ). The inhibitory rates of 12. 5,25.0,50. 0, 100.0 μmoL/L PLE on LECs proliferation were 10.61％ , 14.47％ , 16.40％ and 20. 58％ respectively. Conclusions Polylysine-EDTA can inhibit the growth and proliferation of LECs in vitro at a dose-dependent manner.