进展性大鼠肝纤维化组织中整合素αVβ3受体和血小板内皮细胞黏附分子的表达  

Expression of αVβ3 integrin and platelet-endothelial cell adhesion molecule-I in progressive liver fibrosis： experiment with rats

作　　者：宋正己[1] 王吉耀[1] 涂传涛[1] 王逸清[1] 方国汀[1] 

机构地区：[1]复旦大学附属中山医院消化内科,上海200032

出　　处：《中华医学杂志》2008年第16期1121-1125,共5页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30640054）;上海市科委攻关计划基金资助项目（064119520）

摘　　要：目的探讨整合素αVβ3和血小板内皮细胞黏附分子（CD31）表达与肝纤维化进展和间质重建的关系。方法硫代乙酰胺（TAA）腹腔注射和胆总管结扎（BDL）建立两种大鼠肝纤维化模型，TAA组和BDL组每组8只，分别在第3、9周末和第2,4周末取材。肝组织切片天狼猩红染色和计算机图像分析，整合素αVβ3、CD31免疫化学和荧光染色。整合素αVβ3与α-平滑肌肌动蛋白（SMA）荧光染色双重染色及激光共聚焦观察。实时定量PCB和Western印迹分析肝组织中整合素αVβ3、CD31的相对定量表达。结果肝纤维化中αVβ3表达范围扩大并与α-SMA表达部位一致，CD31在肝血窦、扩大的汇管区和纤维隔表达明显增加，伴有血管新生。模型组和对照组比较肝组织中αVβ3、CD31表达水平差异均有统计学意义（TAA组F=28．66、P〈0．01，F=19．62、P〈0．01和BDL组F=32．60、P〈0．01，F=42．36、P〈0．01）。RAA3周和9周整合素αVβ3和CD31 mRNA的ACt值分别为5．70±0．25、5．50±0．18和4．40±0．25、4．00±0．18。BDL组1周和4周分别为5．60±0．24、5．30±0．14和4．20±0．16、3．80±0．23，表达增加和纤维化进展一致。结论肝纤维化进展中整合素αVβ3表达水平增加与星状细胞活化和血管新生有关，并与间质重建程度平行。Objective To investigate the expression of αVβ3 integrin and platelet endothelial cell adhesion molecuh-1 （ CD31 ） in progressive liver fibrosis of rats. Methods Sixty-four SD rats were randomly divided into 4 equal groups： TAA group, undergoing peritoneal injection of 10% thioacetamine （TAA） 175 mg/kg twice a week to induce liver fibrosis, TAA control group undergoing peritoneal injection of normal saline （ NS）, BDL group undergoing ligation and resection of common bile duct to induce liver fibrosis, and BDL control group undergoing sham operation. Three and 9 weeks later 2 rats from the 2 TAA groups and 1 and 4 weeks later 2 rats from the 2 BDL groups were killed with their liver taken out to undergo sirius red staining and computer image analysis to observe the area of fibrosis, lmmunohistochemistry was used to detect the expression of αVβ3 integrin and CD31. The co-location of αVβ3 integrin and α-smooth muscle antibody （SMA） in the liver tissues was observed by immunohistochemical and double staining. Real-time PCR and Western blotting were used to examine the mRNA and protein expression of αVβ3 integrin and CD31. Results The sirius red stained areas 3 and 9 weeks later of the TAA group were 5.8% ± 1.2% and 16.5% ± 3.6% respectively, and the sirius red stained areas 1 and 4 weeks later of the BDL group were 6.6% ± 1.6% and 18.5% ±4.5% respectively. and double immunotluorescence staining showed that there was an overlapping of αVβ3 and α-SMA expression. Expression of CD31 was extended in the endothelial cells and portal area of both models, and new blood vessels were seen in the fibrotic septum and among the hyperplastic bile ducts. The levels of αVβ3 and CD31 mRNA expression of the TAA and BDL groups were both upregulated （F = 28.66, P 〈 0.01,F=19.62, P〈0.01 and F=32.60, P〈0.01, F=42.36, P〈0.01 respectively） and were increased along with the degree of fibrosis. The levels of protein expression of αVβ3 and CD31 showed the similar trend of their mRNA expression.

关 键 词：肝硬化 整合素ΑVΒ3 抗原 CD31 血管新生 

分 类 号：R686[医药卫生—骨科学]