CCAAT/增强子结合蛋白-α基因对小鼠肝纤维化的治疗作用  

Therapeutic effects of CCAAT/enhancer-binding protein α gene on liver fibrosis in mice

作　　者：陶丽丽[1] 丁笛 张继君[3] 刘秀萍[2] 张锦生[2] 尹为华[1] 

机构地区：[1]北京大学深圳医院病理科,广东省深圳市518036 [2]复旦大学上海医学院病理学系 [3]广东省深圳市第二人民医院病理科

出　　处：《中华消化杂志》2014年第11期737-741,共5页Chinese Journal of Digestion

基　　金：北京大学深圳医院科研基金（201401）

摘　　要：目的 检测CCAAT/增强子结合蛋白-α（C/ EBP-α）基因诱导肝纤维化小鼠体内肝细胞和肝星状细胞（HSC）凋亡的差异.方法 将60只BALB/c小鼠均分为正常组、造模组、治疗组、空白对照组、阴性对照组,每组12只.除正常组外,其余各组腹腔注射四氯化碳（CCl4）建立小鼠肝纤维化模型.治疗组、空白对照组、阴性对照组均于造模第1周经尾静脉分别注射表达C/EBP-α的腺病毒载体（Ad-C/EBP-α）、PBS、腺病毒空载体.免疫组织化学法检测小鼠肝组织中C/EBP-α和α-平滑肌肌动蛋白（α-SMA）的表达.经透射电子显微镜观察小鼠肝组织标本中汇管区周围和远离汇管区部位肝血窦内皮结构.用TUNEL法检测小鼠肝组织的细胞凋亡情况.通过天狼星红染色和肝组织羟脯氨酸含量测定来检测小鼠的肝纤维化情况.多组间比较行单因素方差分析,两组间比较行t检验.结果 正常组C/EBP-α主要表达于肝细胞核内.造模组、空白对照组、阴性对照组C/EBP-α表达量均减少.治疗组C/EBP-α表达量升高,主要表达于肝血窦间隙和血管周细胞.造模组、空白对照组、阴性对照组肝血窦结构扭曲,血管壁增厚,肝细胞变性,出现大量脂滴.治疗组内皮细胞血管壁增厚程度较造模组有所降低.正常组、造模组、治疗组、空白对照组、阴性对照组天狼星红阳性染色细胞所占比例分别为（0.10±0.03）％、（5.81±0.32）％、（2.32±0.45）％、（6.34±0.81）％、（6.10±0.92）％,羟脯氨酸含量分别为（0.07±0.00） μg/mg、（0.69±0.10） μg/mg、（0.19±0.06） μg/mg、（0.56±0.03） μg/mg、（0.64±0.08） μg/mg,α-SMA阳性染色细胞所占比例分别为（0.50±0.03）％、（5.30±0.52）％、（2.15±0.29）％、（5.53±0.43）％、（5.42±0.25）％,TUNEL阳性染色细胞数分别为0.25±0.08、0.15±0.02、7.10±1.53、0.13±0.03、0.18±0.07,各指标组间比较差异有统计学意义（F=113.74、148.2Objective To investigate the difference of CCAAT/enhancer-binding protein α （C/EBP-α） gene induced apoptosis between hepatocytes and hepatic stellate cells （HSC） in mice with liver fibrosis.Methods Sixty BALB/c mice were evenly divided into normal group,model group,treatment group,blank control group and negative control group,12 mice in each group.Except the mice of normal control group,the mice of other groups were treated with intraperitoneal injection of CCl4 to establish liver fibrosis mice model.Mice of treatment group,blank control group and negative control group were administrated with C/EBP-α carried adenovirus （Ad-C/EBP-α）,phosphate buffered solution and empty vector of adenovirus （Ad-EGFP） respectively through tail vein for the first week.The expression of C/EBP-α and α-smooth muscle actin （α-SMA） was detected by immunohistochemistry method.Sinusoidal endothelial structure of peri-portal regions and far from portal regions was observed by transmission electron microscope （TEM）.Terminal dexynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） was applied to detect apoptosis of cells in liver tissue.The degree of liver fibrosis in mice was determined with sirius red staining and hydroxyproline content measurement.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between two groups.Results C/EBP-α was expressed in nucleus of hepatocyte in normal control group mice.The expression decreased in model group,blank control group and negative control group.However,the expression of C/EBP-α of treatment group increased,and mainly expressed in cells located in perisinusoidal and perivascular.Hepatic sinusoids was distorted,blood vessel wall thickened.Hepatocyte degeneration and lots of lipid droplets was found in model group,blank control group and negative control group.The thicken degree of endothelial layer of blood vessel of treatment group was lower than that of model group.The percentage of si

关 键 词：肝硬化 肝星状细胞 CCAAT增强子结合蛋白类 

分 类 号：R575.2[医药卫生—消化系统]