四氯化碳诱导的肝纤维化大鼠肝窦毛细血管化的形成  

The formation of sinusoidal capillarization in the process of liver fibrosis induced by carbon tetrachloride in rats

作　　者：郭杞兰[1] 黄月红[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福州350001

出　　处：《肝脏》2014年第11期839-843,共5页Chinese Hepatology

基　　金：福建省自然基金(青年)(2010J05062);卫生厅青年科研课题(2010-1-10);福建省临床医学重点专科资助项目(闽卫科教[2012]149)

摘　　要：目的观察四氯化碳(CCl4)诱导的大鼠肝纤维化过程中肝窦毛细血管化的形成过程,探讨其与肝纤维化的关系。方法 32只清洁级雄性SD大鼠,随机分为正常对照组,肝纤维化模型组,正常对照组大鼠腹腔注射0.9%氯化钠溶液2 mL/kg,模型组大鼠腹腔注射50%CCl4-蓖麻油混合液2 mL/kg,每周2次,共8周;分别于造模第2、4、6、8周处死大鼠,观察肝组织炎症及纤维化程度,放射免疫法检测血清中透明质酸(HA)的含量,透射电镜观察肝窦窦壁结构,S-P免疫组织化学检测各组大鼠肝组织CD31、层黏连蛋白(LN)、IV型胶原(Col-IV)的表达。结果肝脏组织学证实CCl4诱导的大鼠肝纤维化模型构建成功,6周可见纤维间隔形成;透射电镜显示,CCl4诱导2周时,部分肝窦内皮细胞(liver sinusoidal endothelial cells,LSEC)窗孔减少,内皮下未见基底膜(Basement membrane,BM),随着造模的进程,LSEC窗孔进一步减少,部分甚至消失,第6、8周时局部肝窦内皮下形成连续的BM。同时,随着肝纤维化的进程,HA浓度逐渐升高,肝窦内皮细胞表面标志物CD31及基底膜主要成分Col-IV、LN表达逐渐增强。结论在CCl4诱导大鼠肝纤维化过程中,肝窦毛细血管化是逐渐形成的,LSEC失窗孔早于纤维间隔的形成,而肝窦内皮下基底膜出现在纤维间隔形成以后。Objective To observe the formation of sinusoidal capillarization in the process of liver fibrosis induced by carbon tetrachloride （CCl4 ）in rats,and to explore the relevance between sinusoidal capillarization and hepatic fibrosis. Methods Thirty-two SD rats were divided randomly into control group （group N）and fibrosis model group （group M）. Rats in group N were treated with 2 ml/kg normal sodium twice a week for eight weeks via intraperitoneal injection,while rats in group M were treated with 2 ml/kg a mixture of 50% CCl4 and castor oil twice a week for eight weeks. Rats were executed at the end of 2th wk,4th wk,6th wk,8th wk to collect samples of liver tissue and serum. Grading and staging of hepatic fibrosis were measured by HE staining and masson staining. Changes of HA from serum were measured by radioimmunoassay. Structure of sinus wall in liver sinusoids was observed by TEM. Expressions of CD3 1 ,LN and Col-IV in liver tissue were measured by S-P immunohistochemistry. Results Liver pathohistology analysis confirmed that hepatic fibrosis rat model induced by CCl4 was established successfully. Fibrous septum could be observed at the end of 6th wk. TEM showed that fenestrae of some liver sinusoidal endothelial cells （LSECs）diminished after treated with CCl4 for two weeks with observing no basement membrane （BM）. Fenestrae of LSECs diminished along with the development of hepatic fibrosis,even disappeared. Consecutive BM formed at the end of 6th wk^8th wk in local sinusoidal endothelium. Concentration of HA from serum,and expressions of CD31 ,Col-IV and LN in hepatic sinusoids increased gradually along with development of hepatic fibrosis. Conclusion Formation of sinusoidal capillarization was occurred gradually along with development of hepatic fibrosis. Fibrous septa formed later than fenestrae losing in LSECs,but earlier than formation of sinusoidal endothelial basement membrane.

关 键 词：大鼠 肝纤维化 肝窦毛细血管化 肝窦内皮细胞 基底膜 

分 类 号：R575.2[医药卫生—消化系统]