MCP-1和MIP-2在轮状病毒致新生鼠胆道闭锁中的作用机 .docVIP

MCP-1和MIP-2在轮状病毒致新生鼠胆道闭锁中的作用机制研究 【摘要】 目的：通过轮状病毒毒株（RRV）感染诱导新生鼠胆道闭锁（biliary atresia ，BA)，并检测其肝内单核细胞趋化蛋白-1（MCP-1）和巨噬细胞炎性蛋白-2（MIP-2）的表达情况，探讨其在BA胆管损伤及肝纤维化中的作用。方法：采用腹腔注射猴MMU18006轮状病毒毒株（RRV），建立Balb/c新生小鼠BA动物模型作为实验组，腹腔注射相同剂量的病毒培养液（10%FBS-DMEM）为对照组。在注射后4、7、14及21天各处死一批小鼠，HE染色观察其肝外胆管及肝脏组织学改变，免疫组化法检测MCP-1和MIP-2在肝内的表达，Masson染色观察肝内纤维化程度。应用Image-Pro Plus图像分析软件测定各视野内MCP-1和MIP-2的阳性指数以及肝纤维化程度。结果：实验组小鼠注射病毒后4天出现体重增长缓慢、皮肤黄染等胆道梗阻症状，肝内MCP-1和MIP-2的表达量在4天，7天，14天及21天时均明显高于对照组，其表达水平在注射病毒后14天左右达到高峰，并且两者成正相关（r=0.918，P＜0.001）；Masson染色显示在RRV腹腔注射后4天和7天，小鼠肝脏可见大量胶原蛋白沉积，但未形成明显肝纤维化，14天和21天后，可见肝内纤维化形成，并且胶原蛋白沉积面积与MCP-1表达量成正相关（r=0.529，P=0.005）。结论：BA可能是一种由病毒感染诱发、多种炎症因子介导的免疫性疾病，MCP-1以及MIP-2的大量表达可能在促进BA新生鼠肝内炎症细胞浸润及肝纤维化方面起重要作用。 [关键词]：胆道闭锁，MCP-1，MIP-2，免疫组化，Masson染色 The expression of MCP-1 and MIP-2 in the liver of experimental biliatry atresia 【Abstract】 Objective To explore the function and expression of monocyte chemotactic protein 1 and macrophage inflammatory protein 2 in the liver of biliary atresia (BA) mice. Methods RRV was injected intraperitoneally to induce the mice models of BA that as BA group,and with 10% FBS-DMEM injection as control. Their extrahepatic bile ducts were microdissected 4, 7, 14 and 21 days after injection.Recorded the histopathological changes of the intra- and extrahepatic bile ducts.Then detected the expression of the MCP-1 and MIP-2 by immunohistochemical study and the fibrosis by Masson staining ,and Image-Pro Plus was used to measure the positive index. Result The BA group of mice was significantly slow weight gain , jaundice and other symptoms of biliary atresia . The MCP -1 and MIP -2 expression level of the BA group was significantly higher than the control, and reached a peak about 14 days after injection and there is a positve correlation between the two cytokines(r=0.918,P0.001). When to 4 and 7 days after injection,there are a large number collagen deposition,but did not form a significant liver fibrsis.while intrahepatic fibrosis formed when to 14 and 21 days,an