Proteomic analysis of human tears: defensin expression after ocular surface surgery.
Proteomic analysis of human tears: defensin expression after ocular surface surgery.
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DOI：
10.1021/pr034065n
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发表时间：
2004-03
影响因子：
4.4
通讯作者：
Lei Zhou;Liqun Huang;R. Beuerman;M. Grigg;S. Li;F. Chew;L. Ang;M. Stern;D. Tan
中科院分区：
文献类型：
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作者：
Lei Zhou;Liqun Huang;R. Beuerman;M. Grigg;S. Li;F. Chew;L. Ang;M. Stern;D. Tan
采用表面增强激光解吸/电离飞行时间质谱蛋白质芯片技术（SELDI-TOF ProteinChip）和液相色谱-质谱（LC-MS）对人泪液蛋白质谱进行监测。在手术前（第0天）和术后第1、3和30天，从21例计划手术切除眼表肿瘤的患者中收集泪液。使用这种蛋白质组学方法，我们证实了三种人α-防御素（HNP-1、HNP-2和HNP-3）在手术后的表达中显著上调，并且它们的水平在愈合完成的第30天下降至近似正常。通过LC纯化、胰蛋白酶消化和胰蛋白酶消化产物的ESI-MS/MS分析进一步确认人泪液中α-防御素的身份。测定HNP-1和HNP-2的浓度，并显示在眼表手术后显著增加。研究结果表明，人类α-防御素HNP-1、HNP-2和HNP-3在手术后上调，并且除了它们的抗微生物特性之外，还可能在伤口愈合中发挥重要作用。
Human tear protein profiles were monitored by surface enhanced laser desorption/ionization-time-of-flight mass spectrometry ProteinChip technology (SELDI-TOF ProteinChip) and liquid chromatography-mass spectrometry (LC-MS). Tears were collected from 21 patients scheduled for surgery to remove an ocular surface neoplasm prior to surgery (day 0) and on days 1, 3, and 30 postoperatively. Using this proteomic approach, we verified that three human alpha-defensins (HNP-1, HNP-2, and HNP-3) were significantly up-regulated in their expression after surgery and that their levels decreased to approximately normal by day 30 by which time healing was complete. Further confirmation of the identity of the alpha-defensins in human tears was made by LC purification, trypsin digestion, and ESI-MS/MS analysis of their tryptic digests. The concentrations of HNP-1 and HNP-2 were determined and shown to be markedly increased after ocular surface surgery. The results of the study suggest that human alpha-defensins HNP-1, HNP-2, and HNP-3 are up-regulated after surgery, and may in addition to their antimicrobial properties have an important role in wound healing.