Kent D, Sheridan C, Tomkinson H A, White S, Hiscott P, Grierson I
Unit of Ophthalmology, Department of Medicine, University of Liverpool, Daulby Buildings, Liverpool L69 3GA, London, UK.
Exp Eye Res. 2003 Feb;76(2):213-9. doi: 10.1016/s0014-4835(02)00281-6.
The retinal pigment epithelium (RPE) plays a major role in the development of proliferative vitreoretinopathy (PVR). In particular, RPE cells are implicated in generating the contraction forces seen. The present study was undertaken to investigate whether human RPE binds a lectin from the common edible mushroom, Agaricus bisporus, and to evaluate the effect of any binding on RPE-mediated matrix contraction in an in vitro model of PVR. Fluorescein isothiocyanate (FITC)-labelled Agaricus bisporus lectin (ABL) was used to study binding of lectin to normal retina, PVR scar tissue specimens and cultured human RPE. The effect of a 3-day exposure of ABL on human RPE-mediated contraction was evaluated using 2- and 3D RPE-populated collagen matrices. Effect of ABL on cell adhesion was measured using a collagen type I adhesion assay and determining the relative cellular attachment using absorbance readings. The normal RPE monolayer did not stain with FITC-ABL while PVR scar tissue stained intensely. Staining of in vitro RPE was characteristic but time-dependent. ABL caused a dose-dependent inhibition of RPE-mediated contraction of both 2D (one-way ANOVA, F = 7.94, p < 0.008) and 3D collagen matrices (one-way ANOVA, F = 164.955, p < 0.001). Pre-incubation of ABL with RPE in the 2D model caused a dramatic arrest of contraction (one-way ANOVA, F = 20.1, p < 0.001) that was due to a dose-dependent inhibition of adhesion (one-way ANOVA, F = 15.603, p < 0.001). Recovery of contraction was partially reversible on removal of ABL and was dependent on initial concentration of the lectin. ABL inhibits contraction and adhesion of human RPE cells in vitro without apparent cytotoxicity. It therefore deserves consideration as a potential therapeutic agent in the prevention and treatment of PVR and other non-ocular anomalous wound-healing processes.
视网膜色素上皮(RPE)在增殖性玻璃体视网膜病变(PVR)的发展中起主要作用。特别是,RPE细胞与所观察到的收缩力的产生有关。本研究旨在调查人RPE是否结合来自常见食用蘑菇双孢蘑菇的凝集素,并在PVR的体外模型中评估任何结合对RPE介导的基质收缩的影响。异硫氰酸荧光素(FITC)标记的双孢蘑菇凝集素(ABL)用于研究凝集素与正常视网膜、PVR瘢痕组织标本和培养的人RPE的结合。使用二维和三维RPE填充的胶原基质评估ABL 3天暴露对人RPE介导的收缩的影响。使用I型胶原粘附试验并通过吸光度读数确定相对细胞附着来测量ABL对细胞粘附的影响。正常RPE单层未被FITC-ABL染色,而PVR瘢痕组织染色强烈。体外RPE的染色具有特征性但依赖时间。ABL对二维(单因素方差分析,F = 7.94,p < 0.008)和三维胶原基质(单因素方差分析,F = 164.955,p < 0.001)的RPE介导的收缩均产生剂量依赖性抑制。在二维模型中,ABL与RPE预孵育导致收缩显著停止(单因素方差分析,F = 20.1,p < 0.001),这是由于粘附的剂量依赖性抑制(单因素方差分析,F = 15.603,p < 0.001)。去除ABL后收缩的恢复部分可逆,并且取决于凝集素的初始浓度。ABL在体外抑制人RPE细胞的收缩和粘附,且无明显细胞毒性。因此,它值得作为预防和治疗PVR及其他非眼部异常伤口愈合过程的潜在治疗剂加以考虑。