摘要
目的检测胆道闭锁小鼠模型中肝脏SOX9阳性细胞及SOX9/HNF4α双阳细胞表达特点及其与肝纤维化进展之间的关系。方法取新生的71只BALB/c小鼠进行实验,在出生24~28 h内,新生的57只BALB/c小鼠行20μl RRV腹腔注射作为实验组,其余14只同时注射同体积生理盐水作为对照组,实验组和对照组根据注射后的时间又各自按随机数字表法分为7天组(实验组8只、对照组5只)、10天组(实验组19只、对照组5只)及14天组(实验组30只、对照组4只),观察小鼠临床特征变化。在注射后7 d、10 d、14 d分别留取相应组别小鼠肝脏标本,对肝脏组织切片进行苏木精-伊红染色及天狼猩红染色以评估肝纤维化程度,同时利用多重免疫荧光染色对肝脏组织切片进行SOX9阳性细胞及SOX9/HNF4α双阳细胞计数。利用t检验及One-way ANOVA进行数据分析。结果最终7天组5只小鼠造模成功,10天组6只小鼠造模成功,14天组4只小鼠造模成功,对照组小鼠14只均顺利存活。胆道闭锁小鼠皮肤黄染且体重增长缓慢;所有小鼠肝脏均存在SOX9阳性细胞,胆道闭锁小鼠肝脏中央静脉及小叶间静脉周围SOX9阳性细胞数目随纤维化进展增加,差异具有统计学意义(均P<0.001);对照组小鼠肝脏不表达SOX9/HNF4α双阳细胞,胆道闭锁小鼠肝脏SOX9/HNF4α双阳细胞数量及其占SOX9阳性细胞总数比例随纤维化进展增加,差异具有统计学意义(均P<0.001);SOX9/HNF4α双阳细胞分布与纤维化区域存在重合。结论SOX9/HNF4α双阳细胞在胆道闭锁小鼠肝脏中的表达与纤维化程度进展有关,且SOX9/HNF4α双阳细胞可能参与了胆道闭锁肝纤维化的过程。
Objective To detect the expression patterns of SOX9^(+)cells/SOX9^(+)/HNF4α^(+)cells and examine their relationship with fibrotic stage of livers in a murine model of biliary atresia(BA).Methods A total of 71 neonatal BALB/c mice were selected.In experimental group,57 mice received an intraperitoneal injection of rotavirus;In control group,14 mice had the same volume of physiological saline.Both groups were randomized into 7-day(experimental group,n=8;control group,n=5),10-day(experimental group,n=19;control group,n=5)and 14-day(experimental group,n=30;control group,n=4)based upon injection time.The clinical characteristics were observed.At Day 7/10/14 post-injection,liver samples were harvested.Tissue sections were stained with multiple immunofluorescence for SOX9^(+)and SOX9^(+)/HNF4α^(+)cell count.Sirius red staining was utilized for assessing the degree of liver fibrosis(LF).The relevant clinical data were analyzed by t test and one-way ANOVA.Results Finally 29 mice were obtained(14 mice for control,5 for 7-day,6 for 10-day,4 for 14-day).The skin of BA mice was more jaundiced than control group and their weight grew more slowly;SOX9^(+)cells were detected in all livers;The number of SOX9^(+)cells around central/interlobular vein of liver in BA mice increased with the progression of fibrosis.The difference was statistically significant(all P<0.001);SOX9^(+)/HNF4α^(+)cells were absent in livers of control mice,the number of SOX9^(+)/HNF4α^(+)cells and their proportion in total number of SOX9^(+)cells increased with the progression of fibrosis.The difference was statistically significant(all P<0.0001);The distribution of SOX9^(+)/HNF4α^(+)cells coincided with fibrotic area.Conclusions The number of SOX9^(+)and SOX9^(+)/HNF4α^(+)cells is correlated with the progress of fibrosis and SOX9^(+)/HNF4α^(+)cells may participate in liver fibrosis of BA.
作者
朱佳杰
董瑞
郑珊
Zhu Jiajie;Dong Rui;Zheng Shan(Department of Pediatric Surgery,Children's Hospital of Fudan University,Shanghai Key Laboratory of Birth Defect and Key Laboratory of Neonatal Disease,Ministry of Health,Shanghai 201102,China)
出处
《中华小儿外科杂志》
CSCD
北大核心
2023年第10期936-943,共8页
Chinese Journal of Pediatric Surgery
基金
国家自然科学基金(81974059)
国家儿童医学中心六大临床中心/六大支撑平台建设项目-出生缺陷临床中心(EK2022ZX03)。
关键词
胆道闭锁
肝纤维化
肝细胞核因子
Biliary atresia
Hepatic fibrosis
Hepatocyte nuclear factor
