摘要
目的 :本研究旨在明确正常肝和肝硬化肝细胞中转化生长因子 β1(TGF β1)的作用及其与凋亡的关系。 方法 :通过 5 0 %四氯化碳 (CCl4)腹腔注射诱发BALB/c小鼠形成肝硬化模型 ;应用改良的胶原酶原位灌注法分离正常和肝硬化小鼠的肝细胞 ;利用 1.5 %的琼脂糖凝胶电泳观察DNA梯形条带 ,并应用DNA荧光染料Hoechst33342对正常肝细胞进行染色 ,以检测TGF β1诱导的凋亡。 结果 :以胶原酶原位二步灌流法分离肝细胞的活率为 95 .2 %。正常肝细胞予以TGF β1处理后 ,应用 1.5 %的琼脂糖凝胶电泳可以发现具有凋亡特征的梯形条带。硬化肝细胞则很少出现梯形条带。经TGF β1处理的正常肝细胞应用Hoechst染色发现 ,其凋亡率明显高于未处理组。 结论 :肝硬化肝细胞不能像正常肝细胞那样发生TGF β1诱导的凋亡 。
Objective:The aim of this study was to determine the relationships between the effect of TGF-β 1 and apoptosis in normal and cirrhotic murine hepatocytes. Methods: Liver cirrhosis was induced by means of injection with 50% carbon tetrachloride in BALB/c mice. Hepatocytes were isolated an in situ perfusion method with collagenase solution with minor modification. Normal hepatocytes treated with TGF-β 1(5 ng/ml) were stained with fluorescent DNA-binding dyes Hoechst 33342. The apoptosis of hepatocytes was determined by electrophoresis on 1.5% agarose gel to observe DNA ladder. Results:The viability of hepatocytes isolated from mice by in situ two step collagenase digestion was 95.2%. Internucleosomal fragmented DNA ladder was observed using a 1.5% agarose gel in normal hepatocytes treated with TGF-β 1.obvious DNA ladder from cirrhotic hepatocytes was not observed. The percentage of apoptotic rate in TGF-β 1 treated normal hepatocytes were much higher than that of hepatocytes untreated with TGF-β 1. Conclusion: It was revealed that cirrhotic hepatocytes have poor response to apoptosis induced by TGF-β 1.
出处
《医学研究生学报》
CAS
2004年第10期877-879,共3页
Journal of Medical Postgraduates
关键词
转化生长因子-Β1
凋亡
肝细胞
肝硬化
肝灌注
Transforming growth factor-β 1
Apoptosis
Hepatocyte
Cirrhosis
Liver perfusion
