氮酮对碱性成纤维细胞生长因子眼内通透性的影响

Effect of azone on intraocular permeability of basic fibroblast growth factor

背景研究证实，碱性成纤维细胞生长因子（bFGF）在视网膜前体细胞的增生分化过程中发挥重要作用，但因其眼内通透性低而限制了其在临床上的应用。探寻增加bFGF眼内通透性的有效途径无疑对视网膜疾病的治疗有重要意义。目的观察氮酮对bFGF滴眼液眼内通透性的影响，探索眼部无创给药途径，为bFGF滴眼液用于眼内疾病的治疗提供依据。方法以随机数字表法将18只新西兰白兔随机分成4个组，其中体积分数0．4％氮酮＋体积分数5％bFGF组9只兔并按照取材时间的不同亚分为3个组（30、60、120min），其他3个组各3只兔。不同组兔眼分别局部点用蒸馏水（空白对照组）、5％bFGF滴眼液（5％bFGF组）、0．4％氮酮＋5％bFGF滴眼液（0．4％氮酮＋5％bFGF组）和0．4％氮酮＋10％bFGF滴眼液（0．4％氮酮10％bFGF组），每5分钟1次，共3次，于点眼后30rain抽取房水和玻璃体，其中0．4％氮酮＋5％bFGF组于点眼后30、60和120rain抽取房水和玻璃体，用ELISA法测定房水和玻璃体中bFGF的吸光度（4。）值。结果5％bFGF组兔眼房水和玻璃体中bFGF的4值分别为0．1007-＋0．0100和0．1340＋0．0100，与空白对照组（分别为0．1363±0．0100和0．1130±0．0100）比较影响并不确切。0．4％氮酮＋5％bFGF组及0．4％氮酮＋10％bFGF组兔眼房水和玻璃体中bFGF的A450值均明显高于5％bFGF组，差异均有统计学意义（均P=0．000），但0．4％氮酮＋5％bFGF组与0．4％氮酮＋10％bFGF组房水及玻璃体中bFGF含量的差异均无统计学意义（P=0．985、0．098）。0．4％氮酮＋5％bFGF组点眼后30、60和120min房水中bFGF的A450值以30rain时最高，为0．9413±0．0300，60min时为0．3865±0．0300，120min时为0．2550±0．0300，房水中bFGF的A450值随着时间的延长逐渐降低（R2=0．736，P=0．003），而玻璃体中bFGF的A450值与时间之间无明显线性关系（R2=0．196，P=0．233）。结论氮酮可以改善bFGF滴眼液的眼内通透性，bFGF的体积分数从5％增加到10％时不能有效提高bFGF点眼后的利用度。0．4％氮酮＋5％bFGF滴眼液点眼后30rain房水中bFGF含量最高，随时间延长bFGF的含量逐渐降低。

Background Many researches confirmed that basic fibroblast growth factor （bFGF）plays an important role on the proliferation and differentiation of retinal progenitor cells,but its low intraoeular permeability limits its clinical application. To explore an effective approach to enhance the intraoeular permeability of bFGF has an important significance for the treatment of retinopathy. Objective This study was to investigate the effect of azone on bFGF intraoeular permeability after its topical administration. Methods Eighteen New Zealand white rabbits were randomly divided into four groups on random number table method. Distilled water（ blank control group） ,5% bFGF eyedrops（5% bFGF group）,O. 4% azone＋5% bFGF eyedrops（0.4% azone ＋5% bFGF group） and 0.4% azone＋lO% bFGF eyedrops （0.4% azone ＋ 10% bFGF group）were topically administered in different groups at 5- minute interval for 3 times. Aqueous and vitreous fluid were extracted 30 minutes after administration of eyedrops,and those in the 0.4% azone ＋ 5% bFGF group were obtained 30, 60 and 120 minutes after administration, bFGF concentration in the aqueous and vitreous fluid was quantified with ELISA. Results The bFGF levels（A value）in aqueous and vitreous fluid were 0. 1007±0. 0100 and O. 1340±0. 0100 after topical administration of the 5% bFGF eyedrops,those in blank control group were O. 1363 ± 0. 0100 and 0. 1130 ± 0. 0100, respectively, and those in the 0.4% azone＋5% bFGF group and O. 4% azone＋lO% bFGF group were significantly higher than the 5% bFGF group （ both P = 0. 000 ）. However, no significant difference was found in bFGF levels between 0.4% azone＋5% bFGF group and O. 4% azone＋ 10% bFGF groups in both aqueous and vitreous fluid（ P=O. 985,0. 098）. A value of bFGF in aqueous was gradually increased with prolong of time in the 0.4% azone＋5% bFGF group, with the values 0. 9413 ± O. 0300 at 30 minutes,O. 3865＋0. 0300 at 60 minutes,and O. 2550＋0. 0300 at 120 minutes,showing a positive linear correlation between bFGF level and time（R2 = 0. 736,P=0. 003 ） , but no significant correlation was seen in vitreous sample（R2 = O. 196,P = 0. 233）. Conclusions Azone can improve the intraocular penetration of bFGF eyedrops. Increasing the concentration of bFGF in eyedrop from 5% to 10% dose not change its intraocular distribution. The highest content of the bFGF in aqueous is at 30 minutes following the administration of 0. 4% azone＋5% bFGF eyedrops.