M2BP和M2BPGi在胆道闭锁肝纤维化中的临床意义  

Clinical implications of M2BP and M2BPGi in hepatic fibrosis of children with biliary atresia

作　　者：张聪 赵金凤 郑启鹏 刘更新 杨芳 赵一霖 马慧[3] 胡晓丽[4] 林书祥[5] 詹江华[2] Zhang Cong;Zhao Jinfeng;Zheng Qipeng;Liu Gengxin;Yang Fang;Zhao Yilin;Ma Hui;Hu Xiaoli;Lin Shuxiang;Zhan Jianghua(Graduate School,Tianjin Medical University,Tianjin 300070,China;Department of General Surgery,Tianjin Children's Hospital,Tianjin 300134,China;Department of Laboratory Medicine,Tianjin Children's Hospital,Tianjin 300134,China;Department of Pathology,Tianjin Children's Hospital,Tianjin 300134,China;Pediatric Research Institute,Tianjin Children's Hospital,Tianjin 300134,China)

机构地区：[1]天津医科大学研究生院,天津300070 [2]天津市儿童医院普外科,天津300134 [3]天津市儿童医院检验科,天津300134 [4]天津市儿童医院病理科,天津300134 [5]天津市儿童医院儿研所,天津300134

出　　处：《中华小儿外科杂志》2022年第3期214-220,共7页Chinese Journal of Pediatric Surgery

基　　金：天津市卫生行业重点攻关项目(14KG129);新疆维吾尔自治区自然科学基金(2019D01A12);天津市儿童医院院级课题项目(Y2020002)。

摘　　要：目的本实验旨在进一步验证与评价M2BP与M2BPGi在胆道闭锁(biliary atresia,BA)患儿肝纤维化中的作用。方法收集2019年9月至2020年11月天津市儿童医院普外科收治的48例行肝相关手术的患儿肝组织标本,患儿按疾病类型分为实验组(BA组)35例,其中男17例,女18例,年龄为60(30,60)d;对照组(DC组)13例,其中男7例,女6例,年龄为120(60,720)d。DC组中包括2例胆汁淤积,7例胆总管囊肿(choledochal cyst,CC),4例胆管发育不良(biliary hypoplasia,BH)。通过免疫组织化学检测肝组织中M2BP蛋白含量,分析其与肝组织纤维化分级的相关性;通过实时荧光定量聚合酶链反应(real-time reverse transcription polymerase chain reaction,qRT-PCR),检测肝组织中M2BP mRNA含量,分析其转录、翻译水平是否表达一致。另收集患儿中26例BA、4例BH的血清,检测患儿肝功能相关的生化指标,采用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)检测其血清M2BPGi浓度。患儿组间比较采用Mann-Whitney U秩和检验;用Spearman相关分析评价M2BP及M2BPGi水平与肝纤维化分级的相关性,建立ROC曲线评价血清M2BPGi浓度在肝纤维化中的作用。结果①免疫组织化学结果显示,M2BP表达于肝细胞、巨噬细胞胞质内;BA组患儿肝组织中M2BP蛋白的含量为0.187(0.116,0.222),高于DC组的0.122(0.072,0.141),组间比较,差异有统计学意义(P=0.004);肝组织中M2BPGi的含量与肝纤维化分级存在正相关(rs=0.847,P<0.001)。②qRT-PCR结果显示,BA组患儿肝组织中M2BP mRNA水平为0.099(0.059,0.138),高于DC组的0.036(0.015,0.064),组间比较,差异有统计学意义(P<0.001);Ⅲ~Ⅳ级肝纤维化中M2BP mRNA水平为0.129(0.093,0.194),高于Ⅰ~Ⅱ级肝纤维化的0.060(0.027,0.098),组间比较,差异有统计学意义(P<0.001);与肝组织中M2BP蛋白含量表达一致,随着肝纤维化分级增加,M2BP mRNA水平也随之增加。③ELISA结果显示,Ⅲ~Ⅳ级肝纤维化的血清中M2BPGi含量为6.07(5.0Objective To evaluate the clinical implications of M2BP and M2BPGi in hepatic fibrosis of children with biliary atresia(BA).Methods From September 2019 to November 2020,48 hepatic tissue samples were collected from BA children undergoing hepatic surgery.Experimental group(BA group)was composed of 17 boys and 18 girls with an age range of 26 days to 4 months).And disease control(DC)group had 7 boys and 6 girls with an age range of 60 days to 3 years.The latter group included cholestasis(n=2),choledochal cyst(CC,n=7)and biliary dysplasia(BH,n=4).Immunohistochemistry was employed for detecting the content of M2BP in hepatic tissue and analyzing its correlation with hepatic fibrosis stage.At the same time,real-time reverse transcription polymerase chain reaction(qRT-PCR)was utilized for detecting the mRNA content of M2BP in hepatic tissue to determine whether or not transcription and translation expression levels were consistent.From May 2020 to November 2020,serum samples were harvested from children of BA(n=26),BH(n=4)and cholestasis(n=4).The biochemical parameters of hepatic function at admission were measured and serum M2BPGi level was detected by enzyme-linked immunosorbent assay.Mann Whitney U rank sum test was used for inter-group comparison;Spearman's correlation analysis for evaluating the correlation between the levels of M2BP and M2BPGi and grade of hepatic fibrosis.And receiver operating characteristic(ROC)curve was plotted for evaluating the role of M2BPGi in hepatic fibrosis.Results Immunohistochemical results:M2BP was expressed in cytoplasm of hepatocytes and macrophages.The level of M2BP in hepatic tissues of BA group was 0.187(0.116,0.222).And it was significantly higher than that in DC group 0.122(0.072,0.141)(P<0.001).The content of M2BP in hepatic tissue was strongly and positively correlated with grade of hepatic fibrosis(rs=0.847,P<0.001).qRT-PCR results:mRNA content of M2BP in hepatic tissue BA group was 0.099 ng/ml(0.059,0.138).It was significantly higher than that in disease control group(DC

关 键 词：胆道闭锁 肝纤维化 M2BP蛋白 M2BPGi蛋白 

分 类 号：R725.7[医药卫生—儿科]