METHODS: Normal human keratocytes of corneas from the LIONS Cornea Bank Saar-Lor-Lux, Trier/Westpfalz and human keratocytes of KG and PMD patients were isolated and cultured as keratocytes. To examine Collagen I and V (Col I, Col V), heat shock protein 47 (Hsp47), Lysyl Oxidase (LOX), nuclear factor kappa B (NF-κB) mRNA and protein expression in all cell types, quantitative PCR and Western blot analysis has been performed.
RESULTS: Col5A1 mRNA expression was significantly lower in KG and PMD keratocytes and LOX mRNA expression was significantly higher in KG-keratocytes, compared to controls. Col1A1, Hsp47 and NF-κB mRNA expression and the analyzed protein expressions did not differ from controls, in KG or PMD.
CONCLUSIONS: Col5A1 mRNA expression is decreased in KG and PMD and LOX mRNA expression is increased in KG. Therefore, the pathophysiology of KG and PMD differs from KC and these seem to be from KC independent entities. The explanation of the peripheral corneal thinning in KG and PMD must be investigated in further studies.
方法:正常人角膜细胞,分离并培养KG和PMD患者的Trier/Westpfalz和人角膜细胞作为角膜细胞。检查胶原蛋白I和V(第I卷,ColV),热休克蛋白47(Hsp47),赖氨酰氧化酶(LOX),核因子κB(NF-κB)mRNA和蛋白在所有细胞类型中的表达,已经进行了定量PCR和Western印迹分析。
结果:Col5A1mRNA在KG和PMD角膜细胞中表达显著降低,LOXmRNA在KG角膜细胞中表达显著升高,与对照组相比。Col1A1、Hsp47和NF-κBmRNA表达和分析的蛋白表达与对照组无差异,在KG或PMD。
结论:KG中Col5A1mRNA表达降低,PMD和LOXmRNA表达升高。因此,KG和PMD的病理生理学与KC不同,它们似乎来自KC独立实体。在进一步的研究中,必须研究KG和PMD中周围角膜变薄的解释。