胆管上皮细胞增生在胆管结扎大鼠胆汁性肝纤维化形成中的作用  

The role of cholangiocytes proliferation in the formation of hepatic fibrosis in BDL rats

作　　者：王兵[1] 刘平[1] 龙爱华[1] 陆雄[1] 顾宏图[1] 胡义扬[1] 刘成海[1] 徐列明[1] 

机构地区：[1]上海中医药大学肝病研究所,201203

出　　处：《肝脏》2007年第4期275-279,共5页Chinese Hepatology

基　　金：上海市自然科学基金项目(02JC14024);上海市高校E-研究院基金(E-03008)

摘　　要：目的研究胆管上皮细胞(biliary epitheliac ells,BECs)增生在胆管结扎(bile duct ligation,BDL)大鼠胆汁性肝纤维化形成中的作用。方法BDL的方法制作大鼠胆汁性肝纤维化模型,5周后杀鼠取材,观察内容包括大鼠血清总胆红素(TBlL)、总胆汁酸(TBA),肝组织羟脯氨酸(Hyp)含量,组织病理学,免疫组织化学观测Ⅰ型胶原(ColI)、IV型胶原(ColIV)、层粘蛋白(LN)、纤连蛋白(FN)和α-平滑肌肌动蛋白(α-SMA);肝组织片激光显微切割(LCM)获取BECs,实时荧光定量PCR检测BECs表达Procollagenα1(Ⅳ)、血小板衍生生长因子B(PDGF-B)、结缔组织生长因子(CTGF)及转化生长因子β1(TGF-β1)mRNA。结果1、胆管结扎模型大鼠血清TBiL和TBA含量均值分别为假手术组大鼠的22倍和3倍(P<0.01);模型组大鼠肝组织Hyp含量是假手术组的4倍(P<0.01);2、模型组大鼠BECs增生非常显著,肝实质细胞比例显著减少;3、模型组大鼠在增生胆管周围的LM表达显著增加;FN不但在肝窦内有阳性染色,还强烈地表达于增生的胆管周围;肝窦壁的ColI阳性染色增强,增殖的BECs周围未见明显阳性染色;肝窦壁ColⅣ表达未见明显变化,但强烈表达于增殖的BECs周围的基底膜上;α-SMA阳性染色见于汇管区周围的肌成纤维细胞上,且这些肌成纤维细胞常常围绕在增生的胆管上皮细胞周围。4、模型组大鼠BECs的Procol α1(Ⅳ)、PDGF-B、TGF-β1及CTGF的mRNA表达量均显著增加(与假手术组比较,均为P<0.001)。结论BECs增生是BDL大鼠胆汁性肝纤维化的启动因素和中心病理环节,抑制胆管上皮细胞的异常增生及其细胞生物学病理变化应是抗胆汁性肝纤维化治疗学研究的主要目标。Objective To study the role of biliary epithelia cells （BECs） proliferation in BDL indased hepatic fibrosis in rats. Methods All rats were sacrificed after 5 weeks seram. Samples were obtained and liver tissue hydroproxyline （Hyp） content detected, histopathological examination were performed; Cholangiocytes were obtained by laser capture microdissection（LCM）. Procollagen al （ Ⅳ ）, PDGF-B, CTGF and TGF-β1 mRNA expressions in cholangiocytes were assayed by real time PCR. Results 1. TBiL and TBA content in model group were 22 and 3 folds that of sham operation group（ P 〈 0.01 ） ; 2. Hyp content in model group was 4 folds that of sham operation group （ P 〈 0.01 ） ; 3. Cholangiocytes in model group proliferated significantly and liver parenchymal cells decreased proportionally; LN expression around bile ductule in model group enhanced significantly, FN expressed not only in sinusoidal area, but also around the proliferated biliary duetule. No significant change of Col Ⅳstaining was seen in sinusoidal wall but enhanced expression was seen around basement of proliferating cholangiocytes in model rats, α-SMA staining was seen in fibroblast cells around periportal area in model group, these fibroblast cells usually appeared around cholangiocytes. 4. Procollagen α1（ Ⅳ ）, PDGF-B, TGF-β1 and CTGF mRNA expression in cholangiocytes increased significantly in model group compared to that in sham operation group （ P 〈 0. 001 ）. Conclusion Biliary epithelia cells proliferation is the initiater and central pathological link in BDL hepatic fibrosis in rats. Inhibiting BECs proliferation and cells biological characterestics should be the targets of preventing biliary hepatic fibrosis.

关 键 词：胆管结扎 胆汁性肝纤维化 胆管上皮细胞 增生 

分 类 号：R575.2[医药卫生—消化系统]