目的：利用蛋白芯片技术检测发生与未发生角膜移植排斥反应的大鼠泪液标志蛋白表达,筛选角膜移植排斥反应相关蛋白并分析其与角膜免疫排斥反应相关的可能机制。方法：行大鼠右眼同种异体穿透性角膜移植, I组从角膜旁中心取植片,缝线保留长度约1mm,术后给予抗生素滴眼液；Ⅱ组取角膜中心植片,缝线尽量剪短,术后给予环孢霉素A滴眼液；术后计算排斥反应系数。术后第7d采集泪液。弱阳离子交换芯片结合表面增强激光解析电离飞行时间质谱技术筛选穿透性角膜移植大鼠血清中表达的免疫排斥反应相关的低相对分子质量差异蛋白,结合生物信息学方法初步分析可能的标志蛋白。结果：术后第7d时Ⅰ组发生角膜移植排斥反应,Ⅱ组没有发生；两组中共有108个蛋白质被标记,其中有分类价值的标志蛋白质有11种,相对分类价值较高的有6种,蛋白质搜库结果提示相关蛋白可能是下调蛋白：Pro-histogranin、胰高血糖素样肽1；上调蛋白：β-防御素2、细胞凋亡蛋白酶-3亚基、神经激肽A。结论：应用SELDI-TOF-MS技术获得大鼠角膜移植后泪液标志蛋白表达图谱的实验方法稳定可行,提示我们在角膜的免疫排斥反应的后续研究中考虑能量代谢、神经递质在角膜移植免疫排斥反应中的地位和作用。

AIM:To test the difference of protein composition in tear between cornea transplantation immunological rejection rats and rats with no rejection by SELDI protein chip technology, and to find cornea transplantation immunological rejection associated protein and deduce the possible mechanisms of cornea transplantation immunological rejection.METHODS: To build homogeneity penetrability corneal transplantation rat model: The grafts of group Ⅰ came from side-center and the suture tail remained about 1mm while grafts of group Ⅱ came from center cornea and the suture remained as short as possible. Slit-lamp microscopy was used to observe the three indexes of cornea score in opacity, edema and new vessels from the 1^st day after operation. The result data were as mean±SD and tested by independent-sample ttest in SPSS 13.0 software. The difference was set at P＜0.05. The tears were collected at the 7^th. Capture marker with WCX2 protein chip. Then find out the protein combination that can best distinguish the two group samples. Preliminary screening the marked proteins in Swiss-Prot and TrEMBL protein library.RESULTS: The 7^th day post-operation was sample collection time. It is the time to distinguish the graft rejection group from non-rejection group. 108 proteins were marked in total in two groups, there are 11 kinds of proteins with classified value, among these 6 kinds got high classify value. The result of albumin search library result suggested that the related albumin maybe Neuropeptide Y, β-defensin 1,Glucagon,and Cortistatin-14.CONCLUSION:It is reliable to obtain proteomic profiles in tear for rat marker of cornea graft rejection using SELDI-TOF-MS with well reproducibility. The results suggest that neuripeptide and energy maybe play an important role in graft rejection.