新城疫病毒感染活化的人肝星状细胞系LX-2

摘要

目的 探讨新城疫病毒(NDV)在活化肝星状细胞(HSC)中的复制情况.方法 用转化生长因子β1(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(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莉激活化,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.%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 and RT-PCR was used to measure the changes of related genes mRNA levels. Different passages of LX-2 cells and primary murine 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 HSCs, NDV also had increased replication with HSC passage and TGF-pi stimulation. Conclusion NDV replicate in activated HSC effectively, suggests that activation of HSC facilitate the replication of NDV in this kind of cells.