条件性基因敲除与轮状病毒感染共同作用致小鼠胆道闭锁模型的建立与优化

胆道闭锁是一种预后较差的肝内外胆管闭塞性病变，但发病机制不明。以前采用的胆管结扎或药物注射、基因敲除、轮状病毒感染等方法诱导的动物模型均无法完整模拟人类胆道闭锁的病理改变过程。本研究拟在前期对胆道闭锁病因学研究的基础上，创新性采用基因敲除与轮状病毒感染共同作用的方式诱导小鼠胆道闭锁模型：首先在胆管上皮细胞中敲低与胆管发育相关的基因，并用轮状病毒感染上述细胞，通过评估胆管上皮细胞损伤程度及引发的免疫反应等找到2-3个候选基因；制作条件性基因敲除鼠，对这些基因敲除及对照组新生鼠腹腔注射不同剂量的轮状病毒，观察小鼠生长情况、肝脏病理改变及免疫细胞活化情况，并与人类胆道闭锁的肝脏病理改变相比对，找到与人类胆道闭锁病理改变相一致的轮状病毒感染剂量与基因敲除最佳组合；并研究该动物模型的可靠性和可重复性，从而获得与人类胆道闭锁病理改变相一致的小鼠模型，为开发出减轻胆道闭锁肝脏病损的治疗方法奠定基础。

Biliary atresia (BA) is a progressive obstructive cholangiopathy in intrahepatic and extrahepatic bile ducts with poor prognosis. So far, no individual animal model can mimic the pathological changes of BA perfectly, including bile duct ligation, sclerosing agent injection, gene knockout or rotavirus infection. Based on our previous etiological studies, we aim to establish an innovative murine model of BA by combining key gene knockout and rotavirus infection. Firstly, candidate genes associated with biliary development will be knocked down in.biliary epithelia followed by rotavirus infection, in which 2 to 3 genes will be identified by evaluating the degree of biliary injury and its following immunoreaction. Secondly, we will create or obtain these conditional gene knockout mice, and perform intra-peritoneal injection of various doses of rotavirus in newborn knockout mice and their controls. The growth, hepatic pathological changes and activation of immunocytes will be observed. Finally, we will then compare the hepatic pathological changes between the murine BA and human BA, and we hope to identify the best combination of proper dosage of rotavirus and the most vulnerable candidate genes. After the reliability and repeatability are proven, we hope to acquire a murine BA model which has consistent pathological changes with human BA, which would lay the foundation for future therapies for BA associated hepatobiliary injury.