载5-氟尿嘧啶聚乳酸纳米微粒对人眼球筋膜囊成纤维细胞抑制作用的研究

目的 探讨载5-氟尿嘧啶(5-FU)聚乳酸纳米微粒(5-FU-PLA-NPs)对体外培养的人眼球筋膜囊成纤维细胞的抑制作用.方法 为实验研究.噻唑蓝(MTT)比色法检测不同剂量(0.1、1.0、10.0、100.0和1000.0 mg/L)的空载聚乳酸纳米微粒(PLA-NPs)在不同时间点(48和72 h)对人眼球筋膜囊成纤维细胞增殖的影响,MTT比色法检测不同剂最(0.1、1.0、10.0、100.0和1000.0 mg/L)5-FU纳米药物和5-FU原药在1～7 d内共7个时间点对成纤维细胞生长的抑制作用,计算并比较两者在7个时间点对成纤维细胞的抑制率.RT-PCR分别检测剂量为100.0 mg/L的5-FU纳米药物和5-FU原药作用成纤维细胞1、5、7 d后Ⅲ型前胶原蛋白mRNA的表达情况.对同一天各浓度A值的比较用单因素方差分析,同一天相同浓度5-FU原药组A值与5-Fu-PLA-NPs组A值的比较用两独立样本t检验分析,采用单因素方差分析分别比较不同时间点各组COI3的相对A值.结果 空载聚乳酸纳米微粒对成纤维细胞的增殖没有影响.载5-FU纳米微粒和5-FU原药均抑制成纤维细胞的增殖,并使Ⅲ型前胶原蛋白mRNA的表达水平降低,该抑制作用具有时间和剂量依赖性.作用初期各剂量组5-FU纳米微粒的抑制作用低于原药,随时间延长抑制作用超过原药,差异有统计学意义(P<0.05).结论 聚乳酸(PEA)具有良好的生物相容性与安全性,可作为眼部用药的载体.载5-FU聚乳酸纳米微粒具有一定的缓释功能,可长期释放药物,随着作用时间的延长,5-FU纳米药物对成纤维细胞的抑制作用超过原药,其可作为靶向缓释药物载体.(中华眼科杂志,2009,45:26-31)

The inhibitive effects of 5-fluorouracil-loaded polylactic acid nanoparticles on human Tenon's capsule fibroblast in vitro

Objective To investigate the inhibitive effects of 5-fluorouracil-loaded polylactic acid nanoparticles (5-FU-PLA-NPs)on human Tenon's capsule fibroblasts in vitro. Methods This paper was experimental study. MTT assay was performed to estimate the effects of 0.1 mg/L, 1.0 mg/L, 10.0 mg/L, 100.0 mg/L, 1000.0 mg/L unloaded polylactic acid nanoparticles(PLA-NPs) on fibroblasts proliferation on 48 h, 72 h. MTT assay was performed to estimate the effects of 0.1 mg/L, 1.0 mg/L, 10. 0 mg/L, 100.0 mg/L, 1000.0 mg/L original 5-FU and 5-FU-PLA-NPs on fibroblasts' proliferation on 7 consecutive days respectively. The inhibitory rate of the two drugs against the fibroblasts was calculated. Cells were exposed to 100 mg/L original 5-FU and 5-FU-PLA-NPs respectively for 7 days. Semi-quantitative RT-PCR was used to observe the mRNA expression of type Ⅲ procollagen at the 1st day, 5th day and 7th day. Results PLA-NPs had no cytotoxicity on the fibroblasts. Both 5-FU-PLA-NPs and original 5-FU could inhibit the proliferation of the fibroblasts and make the expression of type Ⅲ procollagen mRNA lower in dose-dependent and time-dependent manner. The inhibitive effect of original 5-FU was more effective than 5-FU-PLA-NPs in the initial period, but the inhibitive effect of 5-FU-PLA-NPs was more effective than original 5-FU in long time. Conclusion PLA has good biocompatibility and safety, and can be used as a carrier of ophthalmic drags. 5-FU-PLA-NPs shows slow-releasing function. 5-FU-PLA-NPs can be proposed as a potential controlled and targeted ophthalmic delivery system for the treatment of antifibrosis after glaucoma filtering surgery. (Chin J Ophthalmol, 2009,45:26-31)