蚕蚀性角膜溃疡患者角膜基质可溶性蛋白和可溶性抗原的初步研究

目的：探讨蚕蚀性角膜溃疡患者角膜基质的可溶性蛋白、可溶性抗原及其血清中抗可溶性抗原的自身抗体变化。方法：采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate polyacrylamide gel electrophoresis technique,SDS-PAGE）方法,检测15例蚕蚀性角膜溃疡患者板层角膜移植术切除的病变角膜及正常人角膜组织中可溶性抗原及可溶性蛋白质浓度,对蛋白电泳条带做薄层扫描定量分析;用间接酶联免疫吸附法（enzyme linked immunoadsorbent assay,ELISA）测定15例患者外周静脉血清中抗角膜基质可溶性抗原的自身抗体含量,并对其结果进行分析。结果：角膜基质提取液的蛋白质浓度：蚕蚀性角膜溃疡组为0．435mg/L,正常对照组为0．645mg/L。角膜基质提取液的SDS－PAGE检测显示：患者角膜基质可溶性蛋白在迁移率为0．92处出现一条对照组未见的蛋白条带,相对分子质量为12000,其占可溶性蛋白总量的22．1％。患者血清中抗角膜基质可溶性抗原的自身抗体含量明显高于对照组（t=15.15,P＜0．001）。结论：蚕蚀性角膜溃疡角膜基质可溶性蛋白相对分子质量为12000,可能与病变角膜基质可溶性抗原有关。患者血清中抗角膜基质可溶性抗原的自身抗体含量异常升高,显示患者的角膜基质发生了自身免疫反应。

A primary study on soluble protein and soluble antigen of corneal stroma in Mooren ' s ulcer patients

OBJECTIVE: To investigate changes of soluble protein and soluble antigen of corneal stroma in Mooren's ulcer patients and autoantibody in the patients' sera against the soluble antigen. METHODS: The corneal stromal specimens taken from 15 Mooren's ulcer patients who received lamellar keratoplasty were studied. Soluble protein and soluble antigen of the corneal stroma were extracted. The soluble protein contents were measured. The soluble corneal stromal extracts were tested by using sodium dodecyl sulfate polyacrylamide gel electrophoresis technique (SDS-PAGE). The protein strips of electrophoresis were analyzed using thin layer scanning system. Peripheral vein blood of 15 patients with Mooren's ulcer was taken. Indirect enzyme linked immunosorbent assay (ELISA) was used to test the level of autoantibody in the patients' sera against the soluble antigen of the corneal stroma. RESULTS: Soluble protein contents of corneal stroma extracts were 0.435 mg/L in the patient group and 0.645 mg/L in the normal control group. SDS-PAGE results of the affected corneal stromal extracts showed that a protein about 12 000 of molecular weight appeared at 0.92 of Rf. 12 000 protein was not found in the control group. The content of 12 000 protein was 22.1% of the total soluble protein of the involved corneal stroma. The level of autoantibody in the patients' sera against the soluble antigen was much higher than that of normal control (t = 15.15,P < 0.001). CONCLUSIONS: The appearance of 12 000 protein in the soluble protein of the involved corneal stroma may have a connection with the soluble corneal stromal antigen. The level of the autoantibody in the patients' sera against the soluble antigen is significantly higher than that of the control. It represents that Mooren's ulcer patients have an autoimmune reaction to the corneal stroma.