豚鼠围产期巨细胞病毒感染肝胆损伤模型的建立  

Establishment of a perinatal cytomegalovirus （CMV） induced hepatobiliary system injury model in guinea pigs

作　　者：王玮[1] 郑珊[1] 

机构地区：[1]复旦大学附属儿科医院外科,上海201102

出　　处：《中华小儿外科杂志》2009年第9期619-625,共7页Chinese Journal of Pediatric Surgery

基　　金：上海市科委重点项目（编号：074119622）;卫生部临床重点项目（编号：卫规财2007353）

摘　　要：目的建立围产期巨细胞病毒感染（CMV）肝胆损伤豚鼠模型，为探讨巨细胞感染对肝胆系统损伤的机制构建载体。方法分组①孕晚期（P组）：分为3组，孕40-43d腹腔接种1×10^9TCID／m1病毒悬液1ml（P1组）、注射生理盐水（P2组）、不注射（P3组）。各组活产子鼠分别于生后12～24h，生后10d，生后20d取血液、肝脏及肝外胆道标本；②新生鼠（N组）：生后12-24h腹腔注射1×10^8TCID）／ml病毒悬液1ml。分别于生后10、20、30d取标本，同前；③幼鼠（Ⅰ组）：生后7～10d腹腔注射1×10^8TCID／ml病毒悬液1ml。分别于注射后10、20、30d处死取标本。血液肝功能检测。肝脏及肝外胆道标本石蜡切片HE染色。部分标本冰冻切片原位杂交检测gpCMV mRNA在肝胆系统的表达。结果①肝功能：孕晚期病毒感染子鼠出生时及生后10dTB、DB、ALT、AST均明显高于对照组（P〈0．05）；至生后20d各指标均明显降低接近正常。P1d10组部分子鼠出现肉眼可见大便变白，伴TB和DB水平的升高。新生鼠病毒接种后10d AST水平较20d组和空白对照升高（P=0．027，P=0．043）；②子鼠生长发育：孕晚期感染子鼠出生体重明显低于对照组（P=0．0029，P=0．0027）。至生后20d体重增长明显落后；③组织病理：63．6％的孕晚期感染小鼠肝脏存在单核淋巴细胞浸润为主的炎症病理改变，小叶间汇管区破坏和纤维化，合并小胆管增生。新生鼠仅生后即刻接种组出现肝脏病变（3／8），但程度较孕晚期组明显减轻。幼鼠组未见明显肝脏病变。gpCMV阳性杂交信号表达于内皮上皮细胞及汇管区基质内，肝细胞内未见表达。孕晚期感染小鼠gpCMV阳性率17．27％，低于新生组和幼鼠组子鼠肝胆系gpCMV-mRNA阳性率（55％，50％，P=0．011〈0．05）。结论孕晚期和出生后即刻腹腔接种未经strain-2传代gpCMV病毒可导致子代豚鼠的感染和肝胆系统Objective To establish a perinatal cytomegalovirus （CMV） induced hepatobiliary system injury model in guinea pigs. Methods Three experimental groups were designed as follows： （1） Prenatal group （Group P）： Female guinea pigs on the 40^th to 43^nt gestational day were randomly allocated into 3 subgroups. Guinea pigs in group P1 and P2 accepted intraperitoneal injection of virus supernatant with the dose of 1 10^9 TCID per dam and saline respectively, while guinea pigs in group P3 served as blank control. Live-born pups were sacrificed within 24 hours, on day 10 or 20 after birth. Samples of livers, extrahepatic bile duct and blood were collected. Weight gain, clinical signs of hepatobiliary injury （i. e. , jaundice in non-fur-covered skin, achoIic stools） and survival were recorded. （2） Neonatal group （Group N） ： A subset of healthy pups accepted intraperitoneal injection of virus supernatant at the dose of 1 10^8TCID per pup within 24 hours after birth. Samples were collected on day 10, 20, or 30 after birth. （3） Infantile Group （Group Ⅰ）： Healthy pups were inoculated with a same dose of virus supernatant on day 10 after birth. Samples were harvested on day 10, 20 or 30 after inoculation. The level of total bilirubin （TB）, direct bilirubin （DB）, ALT and AST in blood samples was analyzed. Serial sections of the liver or extrahepatic bile duct fixed in formalin were stained with Hematoxylin and Eosin. Hybridization in situ was applied on frozen sections to detect the distribution of viral mRNA. Results Compared to those in the control group, TB, DB, ALT and AST levels in Pups infected prenatally were significantly higher within the first 10 days of life （P〈0. 05）, and decreased to the normal level on 20th day of life. A few pups of P1 group got the signs of jaundice （ie.aeholic stools ）, which were associated with increased TB and DB level. In the neonatal group, a higher AST level was observed on the 10th day post inoculation in pups but it returned to nor

关 键 词：肝胆损伤 巨细胞病毒感染 围产期 

分 类 号：R686[医药卫生—骨科学]