重组人内抑素对内皮细胞增殖的影响

目的　探讨原核表达的可溶性重组人内抑素(rhEndo)对ECV 30 4内皮细胞和原代培养兔主动脉内皮细胞增殖的影响。方法　利用酶标仪进行噻唑蓝 (MTT)法检测 ,同时采用倒置相差显微镜、电子显微镜、流式细胞仪、半胱天冬酶 3活性分析观察该内抑素对碱性成纤维细胞生长因子 (bFGF)刺激的血管内皮细胞增殖的影响。结果　该rhEndo明显抑制ECV 30 4细胞的增殖 ,MultiCycleDNACycle软件分析表明细胞增殖阻滞在G1期 ,流式细胞仪检测发现该内抑素可诱导ECV 30 4细胞凋亡 ,且与半胱天冬酶 3活性增强有关 ,但对原代培养的兔主动脉内皮细胞增殖并未产生明显的作用。结论　内抑素可明显抑制ECV 30 4细胞增殖并诱导其发生凋亡 ,但对原代培养的兔主动脉内皮细胞增殖没有明显的影响 ,这将有利于与血管新生有关的疾病如癌症等的治疗。

Effect of recombinant human endostatin on endothelial cell proliferation

AIM To investigate the effects of soluble recombinant human endostatin(rhEndo) expressed in E.coli on the proliferation of ECV 304 and rabbit aortic endothelial cells. METHODS 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method, inverted microscopy, electron microscopy and flow cytometry were used to assay the proliferation of the cells stimulated by basic fibroblast growth factor(bFGF). The caspase 3 activity was also analyzed. RESULTS rhEndo inhibited obviously the proliferation of ECV 304, which was arrested in G₁ phase by analysis with MultiCycle DNA Cycle software. It also induced the apoptosis of ECV 304 which is associated with caspase activity increase, but it did not inhibit the growth of primary culture of rabbit aortic endothelial cells (RAECs). CONCLUSION rhEndo can antagonize the proliferation of endothelial cell line stimulated by bFGF etc., but it may not affect vascular endothelial cells RAECs of primary culture，which is beneficial to the treatment of diseases associated with angiogenesis such as cancer.