急性肝损伤大鼠肝卵圆细胞活化增殖的研究

目的　用D-氨基半乳糖 (D-galactosamine ,D-GalN)联合胆总管结扎 (CommonBiliaryDuctLigation ,CBDL)建立大鼠急性肝损伤模型 ,观察肝损伤后再生过程中肝卵圆细胞 (HepaticOvelCell,HOC)的活化和增殖。 方法　雄性SD大鼠 ,胆总管结扎离断术后予腹腔注射D-GalN 2 .0 g/kg ,对照组不做任何处理。处理后第 1、2、3、4、5及 6天 6个时间点测血清肝功能 ,取肝组织分别行苏木精 伊红染色、电镜、免疫组织化学及RT-PCR检测 ,并对符合HOC形态学特征且胞浆OV-6阳性染色的细胞进行计数。结果　肝功能损害以第 2天明显 (P <0 .0 1) ;对照组肝组织内未见HOC ,处理组各时间点均见有不同程度的HOC增殖反应 ,以第 3天HOC计数最多 (P <0 .0 5 ) ;HOC胞质AFP、OV 6染色阳性 ,胞核BrdU染色阳性 ;RT-PCR示模型组CGT mRNA表达明显增多。结论　用D GalN2 .0 g/kg联合CBDL可获得较满意的大鼠HOC增殖模型 ,在急性肝损伤时HOC被活化和不同程度的增殖 ,参与了肝再生过程

Activation and proliferation of oval cells following acute hepatic damage

Objective To establish a rat model with the combination of D-galactosamine (D-GalN) and common biliary duct ligation(CBDL), and identify the activation and proliferation of oval cells during liver regeneration following hepatic damage.Methods CBDL was performedin in Male Sprague-Dawley rats weighing about 180g each, and then D-GalN was administered intraperitoneally at the dose of 2.0g/kg body weight after the operation. Rats in control group not received any treatment. Three rats were killed every 24 hours after the treatment. A single injection of 5'-bromodeoxyuridine (BrdU) was given intraperitoneally at the dose of 50mg/kg body weight to the animals 1 hour before sacrifice.Blood were drawned for liver function analysis and liver samples were taken for HE staining, electron microscope, immunohistochemistry and RT-PCR evaluation. Three random fields from each slide were counted for the cells with the morphological criteria for oval cells and showing cytoplasmic staining of OV-6. Results Many hepatic oval cells(HOC) were detected in the liver of rats given D-GalN plus CBDL, no HOC were observed in controls. The HOC was characterized by brown staining in the cytoplasm for AFP, OV-6 and in the nucleoli for Anti-BrdU separately. RT-PCR showed expression of GGTmRNA was much higher than the controls.Conclusion Successful rat model for HOC proliferation could be established with D-GalN plus CBDL. HOC could be activated and proliferated following liver damage and it was involved in liver regeneration.