摘要
目的观察白细胞介素(IL)-10基因转染联合γ-干扰素(IFN-γ)对血管平滑肌细胞增殖的影响。方法感染复数(MOI)100的IL-10重组腺病毒(Ad/IL-10)转染人脐带动脉平滑肌细胞(HUASMCs),培养液加200U/ml的IFN-γ,噻唑蓝(MTT)比色法测定吸光度值(A_(570)值)。绘制生长曲线,流式细胞术分析细胞周期及凋亡,实时荧光定量聚合酶链反应(FQ-PCR)检测p21的表达。结果干预第6天,联合组、Ad/IL-10转染组、IFN-γ组的A_(570)值分别为1.342±0.051,1.523±0.012,1.559±0.027,联合组的增殖抑制作用大于单因素组．差异有统计学意义(P＜0.01)；联合抑制呈现时间依赖性；联合组的G_0/G_1期和凋亡细胞数增加,p_(21)表达上调。结论Ad/IL-10转染+IFN-γ是有意义的联合方案。
Objective To investigate the effect of interferon-γ combined with adenovirus-mediated gene transfer of human interleukin-10 on the proliferation of in vitro cultured vascular smooth muscle cells. Methods Human umbilical artery smooth muscle cells （HUASMCs） were in vitro cultured with M199 medium containing epidermal growth factor （EGF） of 40 μg/L end concentration. Multiplicity of infection （MOI） being 100, HUASMCs were transfected with adenoviral vector expressing interleukin-10 （Ad/IL-10）, adding interferon-γ （IFN-γ） of 200 U/ml end concentration into medium. The effect on the proliferation of HUASMCs was investigated by MTT assay and cell grow curves. The cell cycle and apoptosis were also analyzed by using flow cytometry, and the expression of p21 detected by real time fluorescence quantitative polymerase chain reaction （FQ-PCR）. Results At 6th day, the A570 values in combined group, IL-10 transfection group and IFN-γ group were respectively 1. 342 ± 0. 051, 1. 523 ± 0. 012 and 1. 559 ± 0. 027. As compared with groups solely using either Ad/IL-10 or IFN-γ, the proliferation of HUASMCs in combined group was strongly inhibited （P 〈 0.01 ）. In combined group, the cell cycle was significantly arrested in G0/G1 phase, HUASMCs apoptosis was significantly induced, and the p21 expression was upregulated. Conclusion Adenovirus mediated gene transfer of IL-10 plus IFN-γ co-effectively inhibits the proliferation of HUASMCs.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第8期1012-1014,共3页
Chinese Journal of Experimental Surgery
关键词
白细胞介素
干扰素
血管平滑肌细胞
内膜增生
Interleukin
Interferon
Vascular smooth muscle cells
Intimal hyperplasia
