Tong Louis, Li Jing, Chew Jaime, Tan Donald, Beuerman Roger
Singapore National Eye Center, Third Hospital Avenue, Singapore.
Cornea. 2008 Jul;27(6):693-8. doi: 10.1097/QAI.0b013e31815e4041.
Pterygium is a fibro-vascular disease of unknown etiology characterized by proliferation and advancement of tissue onto the cornea. Phospholipase Ds (PLDs) are members of an important class of enzymes involved in inflammation and differentiation. In cultured corneal epithelial cells, these enzymes play a role in wound healing, and in other contexts, they suppress apoptosis and increase cell motility. We aimed to study the presence of PLD subtypes in native ocular surface tissue and pterygium.
This study involved paired control or uninvolved conjunctival and pterygium tissues from 6 patients. Reverse transcription semiquantitative and quantitative polymerase chain reactions were performed to assess transcript levels for PLD1-5 in normal conjunctiva and pterygium tissue. Immunofluorescent staining by using antibodies against PLD1/2 was used to study the expression and tissue distribution. Western blots were performed for protein detection and to confirm the specificity of the antibodies used.
PLD1, 2, 3, and 4 transcripts were detected in normal conjunctiva tissue, and types 2, 3, and 4 were upregulated in pterygium. Immunofluorescent staining showed the presence of phospholipase-D1/2 in normal cornea, conjunctival, and pterygial epithelia. In normal cornea and conjunctival epithelia, the expression was mainly localized to the nuclei of the basal and suprabasal epithelial cells, whereas in pterygium, this expression was limited to the cytoplasm and peri-plasma membrane regions. Western blot confirmed the presence of PLD1/2 in proteins extracted from pterygium and conjunctiva tissue.
PLD subtypes are present in human ocular surface epithelium. PLD may be involved in pterygium pathogenesis.
翼状胬肉是一种病因不明的纤维血管性疾病，其特征是组织向角膜增殖和进展。磷脂酶D（PLD）是参与炎症和分化的一类重要酶的成员。在培养的角膜上皮细胞中，这些酶在伤口愈合中起作用，在其他情况下，它们抑制细胞凋亡并增加细胞运动性。我们旨在研究PLD亚型在天然眼表组织和翼状胬肉中的存在情况。
本研究涉及6例患者的配对对照或未受累结膜及翼状胬肉组织。进行逆转录半定量和定量聚合酶链反应，以评估正常结膜和翼状胬肉组织中PLD1 - 5的转录水平。使用抗PLD1/2抗体进行免疫荧光染色，以研究其表达和组织分布。进行蛋白质印迹检测蛋白质，并确认所用抗体的特异性。
在正常结膜组织中检测到PLD1、2、3和4的转录本，在翼状胬肉中2、3和4型上调。免疫荧光染色显示正常角膜、结膜和翼状胬肉上皮中存在磷脂酶 - D1/2。在正常角膜和结膜上皮中，表达主要定位于基底和基底上层上皮细胞的细胞核，而在翼状胬肉中，这种表达仅限于细胞质和周质膜区域。蛋白质印迹证实从翼状胬肉和结膜组织中提取的蛋白质中存在PLD1/2。
PLD亚型存在于人类眼表上皮中。PLD可能参与翼状胬肉的发病机制。
