rhIFN-α、rhIL-2对血管内皮细胞调控的实验研究

目的：探讨干扰素-α(rhIFN-α)和白细胞介素-2 (rhIL-2) 对人脐静脉内皮细胞增殖、迁移、细胞周期及血管内皮生长因子(VEGF)含量的影响。 方法： 以离体培养的人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVEC)为模型，采用MTT法测定细胞增殖，流式细胞术分析其细胞周期，琼脂糖刮除法检测其对内皮细胞迁移的影响；采用酶联免疫吸附实验(enzyme-linked immunosorbent assay, ELISA)检测上述细胞因子作用下培养的HUVEC血管内皮生长因子（VEGF）在上清液中的含量。 结果： rhIFN-α组细胞增殖和迁移数分别为0.199±0.009和75.750±23.330，rhIL-2组为0.217±0.005和49.250±8.140，而合用组为0.183±0.080和40.500±17.230，与对照组（0.248±0.005和160.500±13.220）比较，有显著差异（均P<0.01）；细胞周期中S期细胞数目，rhIFN-α、rhIL-2单用或合用组分别为14.18±0.55、13.31±0.78、10.05±0.43，而对照组为15.99±1.02（P<0.05或P<0.01）。单独应用rhIFN-α或rhIL-2可显著增加上清液中VEGF含量（P<0.01），而合用组上清液中VEGF水平与对照组比较，差异无显著（P>0.05）。 结论： rhIFN-α、rhIL-2有抑制血管内皮细胞（VEC）增殖、迁移及DNA合成的作用，两细胞因子合用时具有联合效应，提示其在血管生成性疾病的治疗中可能具有一定的作用。

Regulation of rhIFN-α and rhIL-2 on vascular endothelial cell in vitro

AIM: To study the effects of rhIFN-α and rhIL-2 on human umbilical vein endothelial cell (HUVEC) proliferation, migration, cell cycle and vascular endothelial growth factor (VEGF). METHODS: Cultured HUVECs were used as model to determine the cell proliferation with MTT method. Cell cycle was analyzed by cytometry. The cell migration was investigated by agarose scraping method and vascular endothelial growth factor (VEGF) content in supernatant of cultured HUVEC was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The growth and migrating number of endothelial cells in rhIFN-α group were 0.199±0.009 and 75.750±23.330, in rhIL-2 group was 0.217±0.005 and 49.250±8.140, and in combined group was 0.183±0.080 and 40.500±17.230, respectively. In comparison with control group (0.248±0.005 and 160.500±13.220), the effects showed more significant (all P<0.01). By treating the cells with rhIFN-α, rhIL-2 or in combination of both, the cell population in S phase was 14.18±0.55, 13.31±0.78 or 10.05±0.43, respectively and control group was 15.99±1.02 (P<0.05 or P<0.01). VEGF content in supernatant of HUVEC culture in rhIFN-α or rhIL-2 group was higher than those in control group (P<0.01), but VEGF content between in combination group and in the control group was insignificant (P>0.05). CONCLUSIONS: rhIFN-α and rhIL-2 show inhibitory effect on vascular endothelial cell proliferation, migration and DNA synthesis. When used in combination, synergistic effect of rhIFN-α and rhIL-2 is observed, suggesting that these cytokines play an important role in angiogenesis diseases.