Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Homburg/Saar, Germany.
Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Homburg/Saar, Germany.
Exp Eye Res. 2021 Dec;213:108804. doi: 10.1016/j.exer.2021.108804. Epub 2021 Oct 29.
Alike keratoconus (KC), keratoglobus (KG) and pellucid marginal degeneration (PMD) belong to ectatic corneal diseases. While there are numerous studies on keratoconus pathophysiology, there is no exact knowledge on genetic and pathophysiological background of KG and PMD, so far. It is not yet clarified, whether KG and PMD are independent clinical entities or represent different stages of the same disease. Our purpose was to investigate key parameters concerning collagen synthesis, intracellular LOX expression and inflammation in corneal stromal cells of KG and PMD subjects, in vitro.
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.
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.
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.
与圆锥角膜 (KC) 一样,球形角膜 (KG) 和边缘性角膜营养不良 (PMD) 都属于扩张性角膜疾病。虽然有许多关于圆锥角膜发病机制的研究,但迄今为止,对于 KG 和 PMD 的遗传和发病机制背景还没有确切的认识,因此还不清楚 KG 和 PMD 是否是独立的临床实体,或者是否代表同一疾病的不同阶段。我们的目的是研究 KG 和 PMD 患者角膜基质细胞中与胶原合成、细胞内 LOX 表达和炎症有关的关键参数。
从 LIONS 角膜银行 Saar-Lor-Lux、特里尔/西普法尔茨的正常人类角膜中分离并培养正常的角膜基质细胞,并分离培养 KG 和 PMD 患者的 KG 和 PMD 患者的角膜基质细胞。为了检测所有细胞类型中胶原 I 和 V (Col I, Col V)、热休克蛋白 47 (Hsp47)、赖氨酰氧化酶 (LOX)、核因子 kappa B (NF-κB) mRNA 和蛋白的表达,进行了定量 PCR 和 Western blot 分析。
与对照组相比,KG 和 PMD 角膜基质细胞的 Col5A1 mRNA 表达显著降低,而 KG 角膜基质细胞的 LOX mRNA 表达显著升高。KG 或 PMD 中 Col1A1、Hsp47 和 NF-κB mRNA 表达和分析的蛋白表达与对照组无差异。
KG 和 PMD 中的 Col5A1 mRNA 表达降低,而 KG 中的 LOX mRNA 表达增加。因此,KG 和 PMD 的发病机制与 KC 不同,这些似乎是与 KC 无关的实体。在进一步的研究中必须研究 KG 和 PMD 中周边角膜变薄的原因。