大鼠肝移植后移植物胆管中黏蛋白的下调。

Downregulation of mucins in graft bile ducts after liver transplantation in rats.

机构信息

Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, No. 20 Gaotanyan Street, Shapingba, Chongqing, China.

出版信息

Transplantation. 2011 Sep 15;92(5):529-35. doi: 10.1097/TP.0b013e318229f785.

DOI:10.1097/TP.0b013e318229f785

PMID:21775928

Abstract

BACKGROUND

Nonanastomotic biliary strictures represent a serious complication after orthotopic liver transplantation (OLT). This study investigates the potential role of mucins in bile duct injury after OLT.

METHODS

Sprague-Dawley rats were divided into four groups: normal group (Normal, n=5), sham-operated group (Sham, n=20), OLT group with 1 hr donor cold ischemic time (n=20), and OLT group with 12 hr donor cold ischemic time (OLTn=20). Expression of mucins and GATA factors in bile ducts was examined by real-time polymerase chain reaction, immunohistochemistry, and immunoblotting. Bile was collected for biochemical analysis, and the histological changes associated with bile duct injury were evaluated.

RESULTS

In normal bile ducts, Muc1, Muc2, Muc3A, Muc4, and Muc6 mRNA were expressed, whereas Muc5AC mRNA was undetectable. The expression of Muc1, Muc3A, and Muc4 but not Muc2 and Muc6 at mRNA level in graft bile ducts decreased remarkably early after OLT. The decreased expression of Muc1 and Muc4 was further confirmed at protein level by immunohistochemistry and immunoblotting. Downregulation of Muc1 and Muc3A expression by prolonged cold ischemic time was significantly associated with the injury severity scores of large but not small bile ducts. Among six GATA factors, GATA3, GATA4, and GATA6 mRNA were expressed in normal bile ducts. GATA4 and GATA6 mRNA levels decreased significantly after OLT.

CONCLUSION

Downregulation of Muc1 and Muc3A expression by prolonged cold ischemic time may play a potential role in large bile duct injury early after OLT.

摘要

背景

非吻合口胆管狭窄是肝移植（OLT）后的严重并发症。本研究探讨黏蛋白在 OLT 后胆管损伤中的潜在作用。

方法

将 Sprague-Dawley 大鼠分为四组：正常组（Normal，n=5）、假手术组（Sham，n=20）、冷缺血时间 1 小时的 OLT 组（OLT1h，n=20）和冷缺血时间 12 小时的 OLT 组（OLT12h，n=20）。通过实时聚合酶链反应、免疫组织化学和免疫印迹检测胆管中黏蛋白和 GATA 因子的表达。收集胆汁进行生化分析，并评估与胆管损伤相关的组织学变化。

结果

在正常胆管中，Muc1、Muc2、Muc3A、Muc4 和 Muc6mRNA 表达，而 Muc5ACmRNA 无法检测到。OLT 后早期，移植物胆管中 Muc1、Muc3A 和 Muc4mRNA 的表达水平显著降低，但 Muc2 和 Muc6mRNA 的表达水平没有降低。免疫组织化学和免疫印迹进一步证实了 Muc1 和 Muc4 蛋白表达的下调。延长冷缺血时间导致 Muc1 和 Muc3A 表达下调与大胆管而非小胆管损伤严重程度评分显著相关。在 6 种 GATA 因子中，GATA3、GATA4 和 GATA6mRNA 在正常胆管中表达。OLT 后 GATA4 和 GATA6mRNA 水平显著降低。