胆道闭锁异常肝组织蛋白筛查及预后相关性研究

Abnormal expression of hepatic protein and its relationship to prognosis in children with biliary atresia

目的 初步鉴定胆道闭锁肝组织异常蛋白表达情况，寻找与胆道闭锁发病和预后有关的差异表达蛋白。方法 用固相pH梯度双向凝胶电泳分离胆道闭锁和正常肝脏组织总蛋白，银染显色，Melanie3．02 2D图像分析软件分析，对部分蛋白质点用基质辅助激光解析电离飞行时间质谱（MALDI-TOF-MS）进行鉴定，查询数据库鉴定差异蛋白质。结果获得了背景清晰、分辨率和重复性较好的双向凝胶电泳图谱，蛋白质匹配率达80％以上。胆道闭锁和正常组比较发现表达量变化达2倍以上的蛋白点有33个，表达量变化达4倍以上者有8个。胆道闭锁预后较好者和预后不好者比较发现表达量变化达4倍以上的蛋白点有22个，表达量变化达5倍以上者有18个。质谱鉴定其中15个点，其中7个点鉴定成功，分别是Viperin、SARMl、GPC3、APC、THUM2、MIA3和KIAA0649。结论 本研究成功鉴定出部分与胆道闭锁发病和预后有关的蛋白质，为进一步研究胆道闭锁致病机制和提高手术预后提供了新的研究方向，有进一步研究的价值。

Objective To investigate the expression of hepatic protein in patients with biliary atresia（BA）, and find the relationship between the significant protein and prognosis of biliary atresia. Methods Immobile pH gradients isoelectric focusing was used as 1D, and vertical SDS-PAGE as 2D. Sliver staining, Melanie3. 02 2D analysis software, matrix assisted laser desorption ionization-time of flight mass spectrometry （MALDI-TOF-MS） and NCBInr database searching were used to separate and identify the proteome from liver in patients with biliary atresia. Results Satisfactory 2DE patterns were obtained. Thirty-three protein spots were remarkably changed in patients with biliary atresia compared with the control group. Twenty-two protein spots were significantly difference between BA patients with good prognosis and BA patients with poor prognosis. Fifteen protein spots were referred to mass spectrometry, and 7 protein spots were identified, including Viperin, SARM1, GPC3, APC, THUM2, MIA3 and KIAA0649. Conclusions In this study, proteins related to the prognosis and pathogenesis of biliary atresia were identified, which may contribute to prognosis and pathogenesis of biliary atresia.