结膜松弛症患者泪液蛋白质组学研究

目的 比较结膜松弛症和正常人泪液中蛋白质成分的差异,从而探讨结膜松弛症的发病机制.方法 病例对照研究.收集结膜松弛症组和正常对照组各8例(8只眼),采用毛细管从每只眼收集15ul泪液,应用shotgun方法 检测泪液蛋白质,分别行溶液内酶解、反向高效液相色谱(RP-HPLC)分离、电喷雾离子化质谱(ESI-MS/MS)行质谱鉴定后生物信息分析.结果 结膜松弛症组泪液中共有蛋白质356个,正常人泪液中共有蛋白质352个,两组中有119个蛋白质相同.对泪液中蛋白质按基因本体注释(GOA)分类后,发现结膜松弛症组中出现的部分调节凋亡相关蛋白质、凋亡相关蛋白质及炎性反应相关蛋白质在正常对照组中缺失,并且在结膜松弛症泪液中发现了防御素.结论 shotgun方法 可有效分离和分析泪液蛋白质,结膜松弛症患者泪液中的特异性成分提示结膜松弛症可能与细胞凋亡和炎性反应有关.(中华眼科杂志,2009,45:135-140)

The tear protemics analysis of conjunctivochalasis

Objective To analysis tear proteins by using shotgun strategy, and to study pathogenesis mechanism of conjunctivochalasis by comparing tear proteins between the conjunctivochalasis and normal body. Methods Tears were obtained from 8 conjunctivochalasisi cases and 8 normal controls. Fifteen microliters of tears was collected by microcapillary tubes from each eye. Shotgun strategy was used for tear protein analysis. Trypsin digestion in-solution, separation of peptide mixture by reverse-phase high-pressure liquid chromatograph (RP-HPLC) followed by electrosprayionization mass spectrometry (ESI-MS/ MS) identification and bioinformation analysis were used in this study. Results Three hundred and fifty-six proteins were exised in conjunctivochalasis patients and 352 proteins in the normal controls, and among them 119 proteins were the same. The functias of tear proteins were classified with GOA, which found some apoptosis regulation proteins and apoptosis related proteins and inflammatory response related proteins in conjuntivochalasis but not in normal controls group. Defensin was also found in conjuntivochalasis. Conclusion Shotgun strategy can separate and anlysis tear proteins effectively, which provide a new method for tear protein component of conjunctivochalasis. The special component in conjunctivochalasis tear show conjunctivochalasis maybe related to cell apoptosis and inflammatory. (Chin J Ophthalmol, 2009,45:135-140)