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白蛋白前体和热休克蛋白 70 调节器官培养模型中的角膜伤口愈合。

Albumin precursor and Hsp70 modulate corneal wound healing in an organ culture model.

机构信息

Department of Biochemistry, University of Karachi, Pakistan.

出版信息

Acta Histochem. 2011 Jan;113(1):36-42. doi: 10.1016/j.acthis.2009.07.009. Epub 2009 Sep 9.

Abstract

In order to investigate the role of albumin precursor and Hsp70 in corneal wound healing, we have analyzed the distribution of these proteins in wounded and non-wounded corneas of rabbits and the effects of topical applications of anti-albumin precursor and anti-Hsp70 antibodies on wound healing. Anti-albumin precursor and anti-Hsp70 antibodies were topically applied in healing corneal epithelium of rabbit eyes in organ culture. Corneas were allowed to heal in vitro for up to 120 h in serum-free medium with 5 and 10 μg/ml or without (migrating control) anti-albumin precursor/ or anti-Hsp70 antibodies. Fibronectin (Fb) (5 μg/ml) was used as a positive control. Immunofluorescence labelling was used to detect proteins in corneal epithelium at various time intervals following an epithelial defect. Delay in wound healing (p<0.005) was observed with 10 μg/ml anti-albumin antibody labelling. A similar pattern was observed when anti-fibronectin antibody (5 μg/ml) alone and in combination with anti-albumin (10 μg/ml) was ectopically added with wound closure occurring at 120 h. However with anti-Hsp70 antibody (5 μg/ml) slightly delayed (p<0.005) wound closure was observed at 96 h and considerable retardation >120 h with 10 μg/ml. Additionally, immunofluoresence showed a strong co-localization of Hsp70 and albumin precursor during the active phase of wound healing. The presence of albumin precursor and Hsp70 in the epithelial compartment of the cornea indicates a role for these proteins in modulating cell behavior such as epithelial growth, adhesion or regeneration, thus contributing to corneal epithelial wound healing.

摘要

为了研究白蛋白前体和 Hsp70 在角膜伤口愈合中的作用,我们分析了这些蛋白质在受伤和未受伤兔角膜中的分布,以及局部应用抗白蛋白前体和抗 Hsp70 抗体对伤口愈合的影响。在兔眼器官培养的角膜上皮愈合过程中,局部应用抗白蛋白前体和抗 Hsp70 抗体。在无血清培养基中,角膜在体外愈合长达 120 小时,浓度为 5 和 10μg/ml,或无(迁移对照)抗白蛋白前体/或抗 Hsp70 抗体。将纤维连接蛋白(Fb)(5μg/ml)用作阳性对照。在角膜上皮出现上皮缺损后的不同时间间隔,通过免疫荧光标记检测蛋白质。用 10μg/ml 抗白蛋白抗体标记观察到愈合延迟(p<0.005)。当单独使用抗纤维连接蛋白抗体(5μg/ml)和与抗白蛋白(10μg/ml)结合使用时,观察到类似的模式,伤口闭合发生在 120 小时。然而,用 5μg/ml 抗 Hsp70 抗体观察到轻微的愈合延迟(p<0.005),在 96 小时时发生,用 10μg/ml 时则明显延迟>120 小时。此外,免疫荧光显示 Hsp70 和白蛋白前体在伤口愈合的活跃阶段强烈共定位。白蛋白前体和 Hsp70 存在于角膜上皮细胞中,表明这些蛋白质在调节细胞行为方面发挥作用,如上皮生长、粘附或再生,从而有助于角膜上皮伤口愈合。

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