腺病毒介导白细胞介素-10基因转染抑制血管平滑肌细胞的增殖及机制

目的：观察白细胞介素（interleukin，IL-10基因转染对血管平滑肌细胞增殖的影响，初步研究其相关机制。方法：不同感染复数（multiplicity of infection，MOI）的IL-10重组腺病毒（Ad／IL-10）转染原代培养的人脐带动脉平滑肌细胞（HUASMcs），用酶联免疫吸附试验检测培养上清中IL-10的表达，噻唑蓝法测定吸光度值（OD570值），绘制细胞生长曲线，流式细胞术分析细胞周期、细胞凋亡，实时荧光定量聚合酶链反应检测p21的表达。结果：MOI=100时培养上清中IL-10含量最高，为5．746ng／ml；转染72h、96h，MOI为100、150、200组与MOI为10、20组及空白对照组比较，显著抑制HUASMCs的增殖（P〈0．01）；与空白及空载体转染对照组比较，MOI100的Ad／IL-10转染组的Gn-1期细胞数、凋亡细胞的平均百分数均显著增加（P〈0．01），p21基因表达的拷贝数较低。结论：MOI为100的Ad／IL-10可有效抑制HUASMCs的增殖，此作用与细胞周期阻滞和凋亡有关，为ID-10基因转染治疗血管内膜增生提供了理论依据。

Adenovirus Mediated Gene Transfer of Human Interleukin-10 Inhibits Proliferation of Vascular Smooth Muscle Cells and Its Mechanism

Objective:To investigate the effect of adenovirus-mediated gene transfer of human interleukin(IL)-10 on the proliferation of vascular smooth muscle cells, and study related mechanism. Methods: Human umbilical artery smooth muscle cells (HUASMCs) were in-vitro cultured with M199 medium containing epidermal growth factor(EGF) of 40 ng/ml end concentration. After being purified and identified, HUASMCs were transduced with adenoviral vector encoding IL-10 at different multiplicity of infection(MOI). The effect of IL-10 on the proliferation of HUASMCs was investigated by MTT assay and cell grow curves. After transduced at a MOI of 100, we analyzed the cell cycle and apoptosis using flow cytometry and detected the expression of p21 by real time fluorescence quantitative polymerase chain reaction(FQ-PCR). Results: When MOI was 100, IL-10 content in supernatant was highest(5.746ng/ml). At 72 and 96 hours, compared with control group and groups of MOI 10,20 respectively, groups of MOI 100,150,200 significantly inhibited the proliferation of HUASMCs (P<0. 01). Compared with control groups, HUASMCs apoptosis and G0/G1 arrest in group of MOI 100 was significantly induced, and p21 expression was low. Conclusion :Adenovirus mediated gene transfer of IL-10 at a MOI of 100 can effectively inhibit the proliferation of HUASMCs, inhibition is related with G0/G1 arrest and apoptosis and p21 independent.