Wilson S E, Lloyd S A, Kennedy R H
Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas 75235.
Cornea. 1991 Nov;10(6):519-24. doi: 10.1097/00003226-199111000-00010.
Experimental models and clinical investigations have suggested that epidermal growth factor (EGF) may have a role in corneal wound healing. It has been identified as a normal component of human tears. Rabbit and mouse lacrimal glands have recently been shown to synthesize EGF messenger RNA (mRNA). The purpose of the present study was to determine whether the human lacrimal gland synthesizes EGF mRNA. Total cellular RNA was isolated from pathologic specimens of normal human lacrimal glands from two individuals. Reverse transcriptase was used to generate complementary DNA (cDNA) using a human EGF-specific mRNA primer. Amplification of EGF-related cDNA sequences was performed with the polymerase chain reaction (PCR) and human EGF-derived up- and downstream primers. The PCR products from both lacrimal glands contained an amplified product of the expected length of approximately 410 base pairs. The PCR-generated fragment was verified as an EGF-related amplification product with Southern blotting using a synthetic oligonucleotide probe derived from the mature coding sequence of EGF. These results conclusively demonstrate that the human lacrimal gland synthesizes EGF and suggest that the lacrimal gland could have a regulatory role in maintaining the ocular surface and possibly regulating corneal wound healing through the secretion of EGF.
实验模型和临床研究表明，表皮生长因子（EGF）可能在角膜伤口愈合中发挥作用。它已被确定为人类眼泪的正常成分。最近发现兔和小鼠的泪腺能合成EGF信使核糖核酸（mRNA）。本研究的目的是确定人类泪腺是否能合成EGF mRNA。从两名个体的正常人泪腺病理标本中分离出总细胞RNA。使用人EGF特异性mRNA引物，通过逆转录酶生成互补DNA（cDNA）。利用聚合酶链反应（PCR）和人EGF衍生的上下游引物对EGF相关cDNA序列进行扩增。来自两个泪腺的PCR产物均含有预期长度约为410个碱基对的扩增产物。使用源自EGF成熟编码序列的合成寡核苷酸探针，通过Southern印迹法将PCR产生的片段确认为EGF相关扩增产物。这些结果确凿地证明人类泪腺能合成EGF，并表明泪腺可能在维持眼表以及可能通过分泌EGF调节角膜伤口愈合方面具有调节作用。
