用人角膜上皮细胞培养评价5-氟尿嘧啶的眼部不良反应

A. Huhtala, H. Tähti, L. Salminen, H. Uusitalo
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引用次数: 8

摘要

5-氟尿嘧啶(5-FU)常用于眼科青光眼手术后抑制成纤维细胞活性。据报道,对角膜上皮细胞的不良反应与5-FU治疗有关。采用WST-1法(细胞增殖指标)和乳酸脱氢酶(乳酸脱氢酶)法(质膜完整性指标)两种细胞毒性试验,评价5-FU对sv40永生化人角膜上皮细胞(HCE)体外培养的影响。细胞分别在无血清培养基和含15%胎牛血清(FBS)的培养基中暴露于不同浓度的5-FU 1、24、48和72小时。暴露1小时对HCE细胞没有影响。长时间暴露引起剂量依赖性细胞增殖抑制。暴露于5mg /ml 5- fu后,24小时后细胞数量减少到对照组的50%,72小时后导致细胞完全死亡。血清保护细胞24小时,但暴露时间延长后,血清的保护性质消失。5-FU对LDH释放的影响较小。LDH渗漏在处理48h后达到峰值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
EVALUATION OF ADVERSE OCULAR EFFECTS OF 5-FLUOROURACIL BY USING HUMAN CORNEAL EPITHELIAL CELL CULTURES
5-Fluorouracil (5-FU) is commonly used in ophthalmology for suppressing fibroblast activity after glaucoma surgery. Adverse effects on corneal epithelial cells have been reported to relate to 5-FU therapy. The effects of 5-FU were evaluated in vitro on SV40-immortalized human corneal epithelial cell (HCE) cultures with two cytotoxicity tests: WST-1 assay as an index of cell proliferation, and lactate dehydrogenase (LDH) assay as an index of plasma membrane integrity. The cells were exposed to 5-FU with various concentrations in serum-free medium and in medium containing 15 % (v/v) fetal bovine serum (FBS) for 1, 24, 48 and 72 hours. One-hour exposure had no effects on HCE cells. Longer exposures caused dose-dependent inhibition of cell proliferation. Exposure to 5 mg/ml 5-FU lowered cell number to 50 % of controls after 24-hour treatment and resulted to complete cell death after 72 hours. Serum protected the cells for 24 hours, but after longer exposure times the protective nature of serum disappeared. 5-FU had only minor effects on LDH release. The LDH leakage was at its peak after 48-hour treatment.
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