Matsushima Hiroyuki, Mukai Kouichiro, Obara Yoshitaka, Yoshida Shinichiro, Clark John I
Department of Ophthalmology, Dokkyo University School of Medicine, Tokyo, Japan.
J Cataract Refract Surg. 2004 Jan;30(1):187-94. doi: 10.1016/S0886-3350(03)00497-8.
To analyze selected lens cytoskeletal proteins in posterior capsule opacification (PCO) 2 weeks after intraocular lens (IOL) implantation in rabbits.
Department of Ophthalmology, Dokkyo University School of Medicine, Tochigi, Japan.
Eight 10-week-old albino rabbits were prepared and anesthetized for phacoemulsification and aspiration of the crystalline lens and implantation of an acrylic or a hydrogel IOL. Two weeks postoperatively, the rabbits were killed and the IOLs removed for immunohistochemistry. Deparaffinized tissue sections were processed with antibodies against alpha-smooth muscle actin (alpha-SMA) and beta-crystallin to observe the types of PCO with the 2 IOL types. The proteins in the PCO tissue and the normal lens were homogenized, centrifuged, and analyzed using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) densitometric analysis and Western immunoblotting for actin and vimentin.
Immunohistochemistry demonstrated a fibroblastic cell type expressing alpha-SMA and partial regeneration of epithelial cells, resulting in a lenticular structure that stained irregularly for beta-crystallin. The immunoreactivity of fibroblast-like cells to beta-crystallin appeared weaker than that of the regenerated lenticular structure. SDS-PAGE showed variability in the content of cytoskeletal proteins in the insoluble fractions of the PCO. Degradation of the cytoskeletal components was greater with the acrylic IOL than with the hydrogel IOL.
Cytoskeletal proteins expressed during the formation of PCO and IOL implantation may have potential as therapeutic target proteins to improve the biocompatibility of IOLs.
分析兔眼人工晶状体(IOL)植入术后2周后囊膜混浊(PCO)中选定的晶状体细胞骨架蛋白。
日本枥木县独协医科大学医学院眼科。
准备8只10周龄的白化兔,麻醉后行晶状体超声乳化吸除术并植入丙烯酸酯或水凝胶IOL。术后2周,处死兔子,取出IOL进行免疫组织化学检查。对脱石蜡组织切片用抗α平滑肌肌动蛋白(α-SMA)和β-晶状体蛋白的抗体处理,以观察两种IOL类型下PCO的类型。将PCO组织和正常晶状体中的蛋白质匀浆、离心,并用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)密度分析以及肌动蛋白和波形蛋白的蛋白质免疫印迹法进行分析。
免疫组织化学显示一种表达α-SMA的成纤维细胞类型以及上皮细胞的部分再生,导致晶状体结构对β-晶状体蛋白染色不规则。成纤维样细胞对β-晶状体蛋白的免疫反应性似乎比再生的晶状体结构弱。SDS-PAGE显示PCO不溶性部分中细胞骨架蛋白含量存在差异。与水凝胶IOL相比,丙烯酸酯IOL导致的细胞骨架成分降解更大。
PCO形成和IOL植入过程中表达的细胞骨架蛋白可能具有作为治疗靶点蛋白的潜力,以改善IOL的生物相容性。