›› 2009, Vol. 29 ›› Issue (6): 361-365.

• 论文 • 上一篇    下一篇

HGF,EGF对体外培养大鼠肝脏干细胞增殖的实验研究

  

  1. 1.军事医学科学院实验动物中心,北京,100071;2.解放军总医院第一附属医院急救部,北京,100037
  • 收稿日期:2009-04-07 出版日期:2009-06-30 发布日期:2009-06-30

Proliferation Effect of HGF and EGF on Hepatic Stem Cell Cultured from Rat in vitro

  1. Laboratory Animal Center, Military Academy of Medical Sciences, Beijing 100071, China
  • Received:2009-04-07 Online:2009-06-30 Published:2009-06-30

摘要: 目的 研究肝细胞生长因子(HGF)和表皮生长因子(EGF)对体外培养的SD大鼠肝脏干细胞增殖的时间和剂量效应,为肝脏疾病的基础和临床研究奠定基础。方法 肝干细胞用无血清培养基培养,观察不同浓度和不同时间HGF、EGF及HGF和EGF联合对大鼠肝脏干细胞增殖的影响。结果 从成体大鼠肝脏分离的肝干细胞体积小、外形及胞核为圆形或椭圆形.EGF组5~160ng/ml各浓度组、HGF组10~160 ng/ml各浓度组增殖效应均明显大于对照组(P<0.05),且当HGF浓度为20 ng/ml时,增殖效应明显大于与其它各组(P<0.01);HGF组加10 ng/ml EGF组中,除1、5 ng/ml外,其余各组增殖效应均明显大于对照组(P<0.01)。20 ng/ml HGF的时间效应实验结果表明,随着作用时间的延长,对干细胞有明显的增殖效应,于第5天达到高峰,第7天较第5天略有降低.10 ng/ml EGF的时间增殖效应结果显示,随着时间的延长,于第7天达到高峰,但第7天与第5天相比没有显著差异.20 ng/ml HGF和10 ng/ml EGF联合对细胞的增殖效应结果显示,第1天作用不明显,于第3天达到高峰。结论 EGF和HGF具有明显改善SD大鼠肝脏干细胞体外无血清培养条件的作用,对离体培养的大鼠肝脏干细胞增殖具有协同促进作用。

关键词: 肝细胞生长因子, 表皮生长因子, 肝脏干细胞, 增殖

Abstract: Objective To research the effect of time and dosage of offering HGF and EGF comprised of different concentration, which can establish foundation of fundamental and clinical research for liver disease. Methods Hepatic stem cells isolated from portal vein perfusion were cultivated by serum-free medium to observe the effect of HGF and EGF of different concentration and time on proliferation of hepatic stem cell in rat. Results Hepatic stem cells from adult rat have small volume and its outline form and nucleus were round or oval-shape. Proliferation effect in 5—160 ng/ml EGF group and 10-160 ng/ml HGF group was significantly stronger, compared with control (P<0.05); and when HGF was to 20 ng/ml, its effect was the most obvious (P<0.01); In all HGF added with EGF(10 ng/ml) groups, except for 1、5 ng/ml group,proliferation effect were greater than control (P<0.01). In time proliferation effect of hepatic cell treated with HGF (20 ng/ml), with the time prolongation, there exsited manifest effect and was up to the peak at the fifth day; at the seventh day, there happened little decrease. For the time proliferation effect with EGF (10 ng/ml), there emerged the peak at the seventh day. There was no difference at the fifth and seventh day. Proliferation effect of HSC combinated with 20 ng/ml HGF and 10 ng/ml EGF showed that there was not obvious change at the first day; and was up to the peak at the third day. Conclusion There was manifest improvement for the hepatic stem cells cultured by serum-free of SD mouse treated with EGF and HGF. There existed synergistic effect between two factors for the hepatic stem cell proliferation in vitro.

Key words: Hepatocyte Growth Factor, Epidermal Growth Factor, Hepatic stem cell, proliferation