维生素D受体在胆道闭锁患儿肝内胆管上皮细胞中的表达及意义  

Expression level of vitamin D receptor in intrahepatic bile duct epithelial cells and its clinical significance in children with biliary atresia

作　　者：程继文[1] 赵璞 余巧玲 惠军朋 高亚[1] 李鹏[1] Cheng Jiwen;Zhao Pu;Yu Qiaoling;Hui Junpeng;Gao Ya;Li Peng(Department of Pediatric Surgery,the Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China;Department of Neonatology,Shaanxi Provincial People′s Hospital,Xi′an 710068,China;Department of Pathology,the Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China;Department of Pathology,Children′s Hospital Affiliated to Xi′an Jiaotong University,Xi′an 710003,China)

机构地区：[1]西安交通大学第二附属医院小儿外科,西安710004 [2]陕西省人民医院新生儿科,西安710068 [3]西安交通大学第二附属医院病理科,西安710004 [4]西安交通大学附属儿童医院病理科,西安710003

出　　处：《中华实用儿科临床杂志》2022年第10期748-753,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金青年科学基金(81800450);陕西省重点研发计划(S2020-YF-YBSF-0245);西安交通大学自由探索项目(xzy012019087);西安交通大学第二附属医院院人才项目(RC(XM)201801)。

摘　　要：目的探讨肝内胆管上皮细胞(IBDECs)中维生素D受体(VDR)在胆道闭锁(BA)中可能发挥的作用及临床意义。方法回顾性分析2015年1月至2019年12月在西安交通大学第二附属医院和西安交通大学附属儿童医院接受肝门空肠吻合术(Kasai术)治疗的38例BA患儿IBDECs中VDR的表达水平, 免疫组织化学检测VDR在BA患儿IBDECs中的蛋白表达水平, 以胆总管囊肿患儿IBDECs中VDR表达水平为对照;用Masson染色法观察肝组织纤维化程度;并分析BA患儿IBDECs中VDR的表达水平与手术时肝纤维化程度及Kasai术后难治性胆管炎发生率和自体肝生存时间之间的相关性。通过聚肌胞苷酸[Poly(I∶C)]模拟dsRNA病毒感染, 体外干预人肝内胆管上皮细胞(HiBECs), 观察其对HiBECs活性和凋亡的影响及对HiBECs中VDR表达水平的影响。采用独立样本t检验比较两组样本之间的差异;采用χ2检验或Fisher′s确切概率法分析VDR与BA患儿临床病理常数的相关性;采用Kaplan-Meier生存曲线分析不同VDR表达水平组BA患儿Kasai术后自体肝生存时间的差异。结果共38例BA患儿纳入本研究, 其中23例IBDECs中VDR蛋白水平无显著降低, 15例VDR蛋白水平显著降低。与IBDECs中VDR表达水平无明显降低的BA患儿相比, 显著降低者肝纤维化程度更严重(P<0.001), Kasai术后发生难治性胆管炎的比率更高(P=0.017)。与对照组相比, IBDECs中VDR表达水平显著下降的BA患儿自体肝生存时间更短(P=0.030)。Poly (I∶C)可引起BA患儿HiBECs的凋亡率增加(P<0.000 1)、细胞活性降低(P<0.05);Poly (I∶C)还可引起HiBECs培养基中炎症因子(干扰素、肿瘤坏死因子α、白细胞介素-6)分泌增加(P<0.001)。Poly (I∶C)干预可引起BA患儿HiBECs中VDR蛋白的表达水平显著下降(P<0.001)。结论 Poly (I∶C)可引起BA患儿HiBECs损伤及HiBECs中的VDR表达水平下降;IBDECs中VDR表达水平显著下降可能是BA预后不良的标志物。Objective To explore the possible role and clinical significance of vitamin D receptor(VDR)in intrahepatic bile duct epithelial cells(IBDECs)in biliary atresia(BA).Methods A retrospective analysis was performed on expression level of VDR in IBDECs of 38 BA children who underwent Kasai surgery in the Second Affiliated Hospital of Xi′an Jiaotong University and the Children′s Hospital of Xi′an Jiaotong University from January 2015 to December 2019.Expression level of VDR in IBDECs of 38 children with BA was detected by immunohistochemical staining,and that in children with choledochal cysts was detected as negative control.Masson staining was performed to examine the degree of liver fibrosis.The correlation between the expression level of VDR in IBDECs of children with BA,and the degree of liver fibrosis during operation,the incidence of refractory cholangitis after Kasai portoenterostomy and the survival time of autologous liver was analyzed.Human intrahepatic bile duct epithelial cells(HiBECs)were induced with dsRNA virus infection by polyinosinic acid-polycytidylic acid[Poly(I∶C)]in vitro,followed by detection of cell activity,apoptosis and VDR level.The differences between 2 independent groups were analyzed using Student t test.The relationship between the expression of VDR and clinicopathologic characteristics was conducted withχ2 test or Fisher′s test.The Kaplan-Meier survival curve was used to analyze the differences in the survival time of autologous liver after Kasai in BA children with different VDR expression levels.Results A total of 38 children with BA were included in this study.Among them,23 cases showed no significant decrease of VDR protein level in IBDECs,and 15 cases showed a significant decrease in IBDECs.Compared with BA children without a significant decrease in VDR level in IBDECs,much severer liver fibrosis(P<0.001)and significantly higher incidence of refractory cholangitis after Kasai procedure(P=0.017)were detected in those with a significant decrease in VDR level.Compared with

关 键 词：胆道闭锁 维生素D受体 预后标志物 

分 类 号：R726.5[医药卫生—儿科]