Inhibition of latrunculin-A on dexamethasone-induced fibronectin production in cultured human trabecular meshwork cells

AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS: HTM cells were cultured to confluent and incubated with 0.4 mu mol/L Dex and/or 0.05 mu mol/L LAT-A. FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy. RESULTS: Dex up-regulated FN production in HTM cells, failed to do so when co-incubated with LAT-A. LAT-A decreased production of FN in cultured HTM cells. CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma. It has an important prospect as an intraocular pressure-lowering drug.

地塞米松及Lat-A对人眼小梁细胞蛋白质表达的影响

目的探讨地塞米松(Dex)及latrunculin-A(Lat-A)处理后人眼小梁细胞蛋白质表达的变化。方法正常供体人眼小梁细胞培养后用Dex及Lat-A处理,经二维凝胶电泳分离小梁细胞蛋白质,分析凝胶电泳图谱,选取部分差异凝胶斑点并采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)鉴定蛋白质。结果采用24cmIPG胶条获得正常人眼小梁细胞对照组、Dex组、Dex+Lat-A组以及Lat-A组的二维电泳凝胶图谱,得出不同培养条件下人眼小梁细胞蛋白质表达的差异。应用GDPiMALDI-TOFMS得出不同培养条件下差异表达的蛋白质斑点并成功鉴定出其中的24种蛋白质,主要包括细胞骨架相关的蛋白、热休克蛋白、氧化还原相关的蛋白和糖代谢相关的蛋白4类蛋白质。ADLH和Rab蛋白在Lat-A组的人小梁细胞中表达增加,但在Dex组表达减少,而热休克蛋白27(HSP27)和人CRMP2呈现相反的结果。结论成功建立Dex及Lat-A诱导人眼小梁细胞蛋白质表达图谱。Dex及Lat-A能诱导人眼小梁细胞的蛋白质表达变化,可能与青光眼致病的分子机制相关。