胆道闭锁血清抗α-enolase自身抗体检测及临床意义  

Examination of a-enolase autoantibodies in patients with biliary atresia

作　　者：明安晓[1] 王海斌[1] 林海伟[1] 李龙[1] 侯文英[1] 刘树立[1] 张金山[1] 李胜利[1] 刘垚[1] 孙旭[1] 叶茂[1] 

机构地区：[1]首都儿科研究所外科,北京100020

出　　处：《中华小儿外科杂志》2012年第4期263-267,共5页Chinese Journal of Pediatric Surgery

基　　金：基金项目：国家“十一五”科技支撑计划课题（编号：2006BA105A06）;北京市科技计划课题（编号D101100050010064）

摘　　要：目的重组表达人a—enolase蛋白作为抗原，研究胆道闭锁患儿血清中抗a-enolase抗体出现情况及其意义。方法提取肝组织总RNA，逆转录合成cDNA，PCR扩增a—enolase基因全长，构建原核表达质粒，在大肠杆菌中表达并分离纯化重组的a-enolase蛋白。用ELISA检测我实验室于2006年1月至2011年1月收集的胆道闭锁患儿血清标本139例、对照组患儿49例，分析血清中抗a-enolase的IgG或IgM抗体的出现情况。将实施Kasai手术后随访时间大于2年的患儿按ELISA检测（0D值）不同进行分组，比较两组之间6个月内褪黄率、反复胆管炎的发生率以及2年生存率之间的差异，分析该抗体的出现与预后的关系。结果胆道闭锁组OD值〈0．5，0．5～1．0，≥1的例数分别是22例、61例和56例，对照组分别是33例、12例和4例。两者分布差异有统计学意义（P〈0．05）。Kasai术后随访时间大于2年的患儿共78例，按照OD值的不同分为2组（〈0．85和〉0．85），两组患儿6个月内退黄率分别是47％和80％，两组差异有统计学意义（x2=9．018，P=0．003）；反复胆管炎的发生率为58％和30％，两组差异有统计学意义（x2=6．617，P=0．013）；2年生存率分别是47％和60%，两组差异无统计学意义（x2=1．251，P=0．263）。结论部分胆道闭锁患儿血清中有抗a—enolase的IgG或IgM抗体出现，针对a-enolase的体液免疫反应可能参与了胆道闭锁的发生过程。抗体水平的升高有利于短期预后的恢复，对长期预后的影响仍需进一步确认。Objective To produce and purify recombinant human a-enolase protein, and to exmaine the level of a-enolase autoantibodies in serum of the patients with biliary atresia （BA）. Methods The eDNA sequence of humana-enolase obtained by PCR was cloned into prokaryotie expression vector PET41-a. PET41-a was then transfeeted into E. coli BL21. The a-enolase protein expressed in E. eoli BL21 was isolated and purified. During January 2006 to January 2011, 139 patients with BA and 49 healthy controls were recruited in this study. Their serum were collected, and IgG and IgM auto-antibodies to a-enolase were studied by ELISA. The patients who survived 2 years after Kasai procedure were grouped into 2 groups according to the level of IgG and IgM auto-antibodies to a-enolase in serum. The clinical data of the 2 groups was studied and compared. Results In BA group, there were 22 patients＇ OD value less than 0. 5, 61 between 0. 5-1, and 56 more than 1. In healthy controls, there were 33 patients＇ OD value less than 0. 5, 12 between 0. 5-1, and 4 more than 1. The OD value was statistieally different between BA group and controls （P〈0. 05）. Seventy eight patients survived 2 years after Kasai procedure. The patients were divided into 2 groups according to whether or not their OD value reached 0. 85. The postoperative jaundice clearance was significantly better in the patients whose OD value 〉0. 85 compare with those whose OD value 〈0. 85 （80% vs 47%, x2 = 9. 018, P = 0. 003）. The recurrent cholangitis was significantly lower in the patients whose OD value 〉0. 85 compare with those whose OD value 〈0. 85 （30% vs 58%, x2 = 6. 617, P= 0. 013）. The 2-year postoperative native liver survival rate was significantly better in the patients whose OD value 〉0. 85 compare with those whose OD value 〈0. 85 （60% vs 47%, Z2 = 1. 251, P = 0. 263）. Conclusions Most of the BA patients have high level of a-enolase autoantibodies in serum. The level of the autoantibody was positively correlated with short-term outco

关 键 词：胆道闭锁 自身抗体 

分 类 号：R726.5[医药卫生—儿科]