Qu Mingli, Wang Yao, Yang Lingling, Zhou Qingjun
State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.
Mol Vis. 2011;17:3147-55. Epub 2011 Dec 6.
To evaluate and compare the cellular effects of four commercially available anti-inflammatory eye drops and their active components on human corneal epithelial cells (HCECs) in vitro.
The cellular effects of four eye drops (Bromfenac Sodium Hydrate Eye Drops, Pranoprofen Eye Drops, Diclofenac Sodium Eye Drops, and Tobramycin & Dex Eye Drops) and their corresponding active components were evaluated in an HCEC line with five in vitro assays. Cell proliferation and migration were measured using 3-(4,5)-dimethylthiahiazo (-z-y1)-3 5-di-phenytetrazoliumromide (MTT) assay and transwell migration assay. Cell damage was determined with the lactate dehydrogenase (LDH) assay. Cell viability and median lethal time (LT₅₀) were measured by 7-amino-actinomycin D (7-AAD) staining and flow cytometry analysis.
Cellular effects after exposure of HCECs to the four anti-inflammatory eye drops were concentration dependent. The differences of cellular toxicity on cell proliferation became significant at lower concentrations (<0.002%). Diclofenac Sodium Eye Drops showed significant increasing effects on cell damage and viability when compared with the other three solutions. Tobramycin & Dex Eye Drops inhibited the migration of HCECs significantly. Tobramycin & Dex Eye Drops showed the quickest effect on cell viability: the LT₅₀ was 3.28, 9.23, 10.38, and 23.80 min for Tobramycin & Dex Eye Drops, Diclofenac Sodium Eye Drops, Pranoprofen Eye Drops, and Bromfenac Sodium Hydrate Eye Drops, respectively. However, the comparisons of cellular toxicity revealed significant differences between the eye drops and their active components under the same concentration. The corneal epithelial toxicity differences among the active components of the four eye drops became significant as higher concentration (>0.020%).
The four anti-inflammatory eye drops showed different cellular effects on HCECs, and the toxicity was not related with their active components, which provides new reference for the clinical application and drug research and development.
评估并比较四种市售抗炎眼药水及其活性成分对人角膜上皮细胞(HCECs)的体外细胞效应。
采用五种体外检测方法,在HCEC系中评估四种眼药水(布罗芬酸钠水合物眼药水、普拉洛芬眼药水、双氯芬酸钠眼药水和妥布霉素地塞米松眼药水)及其相应活性成分的细胞效应。使用3-(4,5)-二甲基噻唑-2,5-二苯基四氮唑溴盐(MTT)检测法和Transwell迁移检测法测量细胞增殖和迁移。用乳酸脱氢酶(LDH)检测法测定细胞损伤。通过7-氨基放线菌素D(7-AAD)染色和流式细胞术分析测量细胞活力和中位致死时间(LT₅₀)。
HCECs暴露于四种抗炎眼药水后的细胞效应呈浓度依赖性。在较低浓度(<0.002%)时,细胞毒性对细胞增殖的差异变得显著。与其他三种溶液相比,双氯芬酸钠眼药水对细胞损伤和活力有显著增加的作用。妥布霉素地塞米松眼药水显著抑制HCECs的迁移。妥布霉素地塞米松眼药水对细胞活力的作用最快:妥布霉素地塞米松眼药水、双氯芬酸钠眼药水、普拉洛芬眼药水和布罗芬酸钠水合物眼药水的LT₅₀分别为3.28、9.23、10.38和23.80分钟。然而,细胞毒性比较显示,在相同浓度下,眼药水与其活性成分之间存在显著差异。四种眼药水活性成分之间的角膜上皮毒性差异在较高浓度(>0.020%)时变得显著。
四种抗炎眼药水对HCECs显示出不同的细胞效应,且毒性与其活性成分无关,这为临床应用及药物研发提供了新的参考。