脂质体阿霉素治疗实验性增生性玻璃体视网膜病变时胶质反应减弱。

Attenuated glial reaction in experimental proliferative vitreoretinopathy treated with liposomal doxorubicin.

机构信息

Department of Ophthalmology, Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, Taiwan.

出版信息

Invest Ophthalmol Vis Sci. 2012 Jun 29;53(6):3167-74. doi: 10.1167/iovs.11-7972.

DOI:10.1167/iovs.11-7972

PMID:22491400

Abstract

PURPOSE

We investigated the therapeutic effect of liposomal doxorubicin (Lipo-dox) on experimental proliferative vitreoretinopathy (PVR).

METHODS

The toxicity of Lipo-dox was determined in vitro in cultured rabbit retinal pigment epithelium (RPE) cells by tetrazolium-based (MTT) assay for cell viability performed 48 and 96 hours after treatment, and in vivo by electroretinography and histopathology. The therapeutic effect of intravitreous injection of Lipo-dox was evaluated in a rabbit model of PVR induced by injection of rabbit RPE cells after gas compression of the vitreous. The presence of PVR was determined by indirect ophthalmoscopy on days 1, 7, 14, 21, and 28 after injection. Western blot and immunofluorescence studies were performed to evaluate the expression of the glial markers vimentin and glial fibrillary acidic protein (GFAP). A pharmacokinetic study also was performed and analyzed by liquid chromatography coupled with tandem mass spectrometry (LC-MS).

RESULTS

The 50% inhibitory concentrations (IC₅₀) of doxorubicin (Doxo) and Lipo-dox in RPE cells were 0.01-0.1 and 0.1-1.0 μg/mL, respectively. Lipo-dox (10 μg/mL) did not reduce the amplitude reduction in the ERG study or produce obvious retinal toxicity. Lipo-dox still could be detected in the vitreous 7 days after injection. The Lipo-dox (10 μg/mL)-treated eyes showed lower grade PVR than did the untreated eyes. Lipo-dox also decreased the retinal expression levels of vimentin and GFAP.

CONCLUSIONS

Lipo-dox can attenuate the severity of experimental PVR, and reduces the glial cell expression of intermediate filaments in PVR retinas. Lipo-dox has a wider safe dosage range and a longer half-life in the vitreous than does primary Doxo.

摘要

目的

研究脂质体阿霉素（Lipo-dox）对实验性增生性玻璃体视网膜病变（PVR）的治疗作用。

方法

通过四唑盐（MTT）比色法测定细胞活力，在体外培养的兔视网膜色素上皮（RPE）细胞中测定 48 和 96 小时后 Lipo-dox 的毒性，以及通过视网膜电图和组织病理学进行体内毒性测定。通过向玻璃体内注入兔 RPE 细胞后进行气体压缩，在兔 PVR 模型中评估玻璃体内注射 Lipo-dox 的治疗效果。通过间接检眼镜在注射后第 1、7、14、21 和 28 天确定 PVR 的存在。通过 Western blot 和免疫荧光研究评估神经胶质标志物波形蛋白和神经胶质纤维酸性蛋白（GFAP）的表达。还进行了药代动力学研究，并通过液相色谱-串联质谱（LC-MS）进行分析。

结果

阿霉素（Doxo）和 Lipo-dox 在 RPE 细胞中的 50%抑制浓度（IC₅₀）分别为 0.01-0.1 和 0.1-1.0 μg/mL。Lipo-dox（10 μg/mL）不会降低 ERG 研究中的振幅降低，也不会产生明显的视网膜毒性。注射后 7 天，Lipo-dox 仍可在玻璃体内检测到。Lipo-dox（10 μg/mL）治疗的眼睛比未治疗的眼睛的 PVR 分级低。Lipo-dox 还降低了视网膜中波形蛋白和 GFAP 的表达水平。