基础医学与临床 ›› 2012, Vol. 32 ›› Issue (10): 1161-1166.

• 研究论文 • 上一篇    下一篇

新城疫病毒感染活化的人肝星状细胞系LX-2

李亚琳   

  1. 渭南师范学院
  • 收稿日期:2011-10-24 修回日期:2012-03-05 出版日期:2012-10-05 发布日期:2012-09-28
  • 通讯作者: 李亚琳 E-mail:lyal1222@126.com
  • 基金资助:
    新城疫病毒抑制肝星状细胞活化的体内外研究;CD147调节肝星状细胞活化及其逆转肝癌发生发展的溶瘤病毒治疗学基础研究;新城疫病毒抑制小鼠肝纤维化的机制研究

Newcastle disease virus infect activated human hepatic stellate cells LX-2

YALIN LI   

  • Received:2011-10-24 Revised:2012-03-05 Online:2012-10-05 Published:2012-09-28
  • Contact: YALIN LI E-mail:lyal1222@126.com

摘要: 目的 探讨新城疫病毒(Newcastle disease virus, NDV)在活化肝星状细胞(Hepatic stellate cell, HSC)中的复制情况。方法 用转化生长因子β1(Transforming growth factor, TGF-β1)刺激人肝星状细胞LX-2,MTT法检测细胞的增殖率, RT-PCR检测活化相关基因的mRNA表达。用NDFLtag-EGFP感染不同传代的LX-2细胞和原代分离培养以及TGF-β1刺激活化的HSC,荧光显微镜观察NDV在细胞中的复制。用流式细胞术(Flow cytometry, FACS)检测NDFLtag-EGFP和NDV-Italien在LX-2细胞中的复制。结果 TGF-β1能够刺激LX-2细胞的活化。NDV随着LX-2细胞的连续传代活化以及TGF-β1刺激活化,其在细胞中的复制率也逐渐增加,分别从(15.65±0.92)%增加到(23.05±1.5)%、从 (12.8±1.4)% 增加到(22.7±1.7)%,(P<0.05)。原代分离培养的小鼠HSC,随着细胞的传代次数增加和TGF-β1的刺激,NDV也表现出较高的复制效率。结论 NDV能在活化的HSC中高效复制, LX-2的活化易化了NDV在其中的复制。

关键词: 新城疫病毒, 肝星状细胞, 转化生长因子β1, 活化

Abstract: Objective To explore the replication efficiency of Newcastle disease virus in hepatic stellate cells. Methods Human hepatic stellate cells LX-2 were stimulated with TGF-β1 and the proliferation of LX-2 cells was detected by MTT assay, RT-PCR was used to measure the changes of related genes mRNA levels. Different passages LX-2 cells and primary mouse HSCs with or without TGF-β1 stimulation were infected by NDFLtag-EGFP, NDV replication in these cells was observed under fluroscent microscopy. The replication of NDFLtag-EGFP and NDV-Italien in LX-2 was detected by flow cytometry (FACS). Results TGF-β1 stimulated the activation of LX-2 cells. The efficiency of NDV replication was increased with the consecutive passages (from (15.65±0.92)% to (23.05±1.5)%, (P<0.05)) and TGF-β1 stimulation of LX-2 cells (from (12.8±1.4)% to (22.7±1.7)%, (P<0.05)). In primary isolated mouse HSCs, NDV also had increasing replication with HSC passage and TGF-β1 stimulation. Conclusions NDV replicate in activated HSC effectively, it means that activation of HSC facilitate the replication of NDV in this kind of cells.

Key words: Newcastle disease virus, Hepatic stellate cell, TGF-β1, activation