摘要
目的:研究骨桥蛋白(OPN)及纤溶酶原激活物抑制物(PAI-1)的表达特征及其在肝纤维化时的变化.方法:采用二甲基亚硝胺制作大鼠肝纤维化模型.大鼠肝脏常规HE和天狼猩红染色.采用SABC法作免疫组织化学染色及Westernblot检测OPN和PAI-1蛋白表达;RT-PCR法检测OPN基因表达;检测结果采用计算机图像定量分析系统,扫描并计算染色阳性区域面积和阳性比率或条带的吸光度值.结果:正常大鼠肝组织OPN和PAI-1表达极弱,肝纤维化大鼠肝脏中OPN表达增强,阳性信号散在或弥漫性分布,主要见于小叶内中央静脉周围、纤维间隔内以及周围巨噬细胞胞质,库普弗细胞,门管区的部分肝细胞,肝窦壁内皮细胞.PAI-1在肝纤维化大鼠肝组织汇管区、肝细胞变性坏死处,肝窦周Disse间隙及毗邻以上部位的肝细胞,组织纤维间隔处及其外周细胞亦见阳性染色.Westernblot检测正常大鼠肝脏OPN的蛋白表达极低,肝纤维化组OPN的蛋白表达较正常组显著增强(1.0907±0.2082vs0.0673±0.0663,P<0.01).与正常组比,肝纤维化组PAI-1表达也显著增强(1.1407±0.3094vs0.2464±0.2234,P<0.01).RT-PCR检测结果显示,正常大鼠肝脏OPNmRNA表达极低,肝纤维化大鼠肝脏OPNmRNA的表达明显增强(0.2128±0.0527vs0.1298±0.0316,P<0.05).结论:OPN及PAI-1的表达与肝纤维化密切相关,肝纤维化时大鼠肝组织OPN及PAI-1的表达水平显著增高,OPN可能会促进PAI-1的高表达,从而抑制细胞外基质(ECM)降解、加速肝纤维化进程.
AIM： To investigate the expression of osteopontin （OPN） and plasminogen activator inhibitor type 1 （PAI-1） in dimethylnitrosamine （DMN）-induced hepatic fibrosis in rats. METHODS： Liver fibrosis was induced in rats by intraperitoneal injection of DMN. All rats were sacrificed to take liver tissue specimens. Formalin-fixed liver sections were stained with hematoxylin and eosin （HE） and Sirius Red to evaluate general histology and hepatic fibrosis, respectively. The expression of OPN and PAI-1 proteins was detected by immunohistochemistry and Western blot. The expression of OPN mRNA was detected by reverse transcriptionpolymerase chain reaction （RT-PCR）. Computerized image analysis system was then used for quantification. RESULTS： Immunohistochemical stainingshowed that the expression of OPN and PAI-1 increased dramatically in fibrotic rat liver when compared with normal rat liver. OPN-positive signals were scattered or diffused around the central veins, and in fibrous septum and adjacent macrophages, Kupffer cells, some hepatic cells in the portal area, and liver sinusoidal endothelial cells. The expression of PAI-1 was distributed in the portal areas, degenerative and necrotic hepatic cells, the space of Disse and adjacent hepatic cells, fibrous septum and adjacent cells. Western blot analysis showed that the expression of OPN and PAI-1 increased obviously in fibrotic rat liver （1.0907 ± 0.2082 vs 0.0673 ± 0.0663 and 1.1407 ± 0.3094 vs 0.2464 ± 0.2234, respectively; both P〈0.01）. RT-PCR analysis showed that the expression of OPN mRNA was also significantly enhanced in fibrotic rat liver （0.2128 ± 0.0527 vs 0.1298 ± 0.0316, P〈0.05）. CONCLUSION： The expression of OPN is closely correlated with that of PAI-1 during hepatic fibrosis. The expression of OPN and PAI-1 is significantly up-regulated in rat hepatic fibrosis. OPN may promote the high expression of PAI-1, inhibit extracellular matrix degradation, and accelerate fibrosis progression.
出处
《世界华人消化杂志》
CAS
北大核心
2010年第11期1104-1108,共5页
World Chinese Journal of Digestology
基金
上海市中医肝病临床医学中心基金资助项目
No.ZX2006-01~~
关键词
骨桥蛋白
纤溶酶原激活物抑制物
肝纤维化
肝组织
Osteopontin
Plasminogen activator inhibitor type-1
Hepatic fibrosis
Hepatic tissue
