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目的建立稳定的人脐带血管内皮细胞(ECs)和平滑肌细胞(SMCs)模型研究霉酚酸(MPA)对SMCs增殖和胶原Ⅰ产物的作用。方法以北京大学航天临床医学院2002-09～2003-09从脐带静脉、动脉分别获取和培养ECs和SMCs;用无血浆DMEM细胞培养液制备含或不含MPA(质量浓度分别:0、0.31、1.25、2.50、5.00、10.00μg·mL-1)的ECs调节的细胞培养液;用细胞与3H-胸腺嘧啶结合闪烁记数法测定SMCs增殖;用酶联免疫法测定SMCs释放到培养液中的胶原之量。结果与对照组相比,在ECs调节的培养液(不含MPA)作用下,实验组SMCs增殖显著提高到(170.6±13.7)%(P<0.01),胶原Ⅰ产物显著增加到(128.0±7.7)%(P<0.01);对非刺激状态SMCs,MPA只在高质量浓度(10μg·mL-1)时直接显著地抑制SMCs增殖(P<0.05),抑制率(29.9±3.5)%。在所有实验浓度,MPA呈剂量依赖地显著抑制SMCs胶原Ⅰ产物,最大抑制率(81.0±4.8)%;对在ECs调节的培养液作用下,无论是在ECs调节的培养液制备过程中加入MPA作用于ECs,还是在ECs调节的培养液制备后加MPA到培养液,MPA都明显地抑制SMCs增殖和胶原Ⅰ产物。结论MPA能够一定程度地抑制人血管SMCs增殖,同时能很好地抑制SMCs胶原Ⅰ产物。
Objective To establish a stable model of human umbilical vascular endothelial cells (ECs) and smooth muscle cells (SMCs) to study the effects of mycophenolic acid (MPA) on proliferation and collagen Ⅰ production in SMCs. Methods ECs and SMCs were harvested and cultured from the umbilical vein and artery of Peking University School of Astronautics and Clinics of Peking University from September 2002 to September 2003, respectively. MPA was prepared with or without MPEM (concentration: 0,0.31, 1.25, 2.50, 5.00 and 10.00μg · mL-1). The proliferation of SMCs was measured by scintillation counting with 3H-thymidine and the release of SMCs into culture medium by enzyme-linked immunosorbent assay The amount of collagen. Results Compared with the control group, the proliferation of SMCs in experimental group was significantly increased to (170.6 ± 13.7)% (P <0.01) under ECs conditioned medium (without MPA), and the product of collagen Ⅰ significantly increased to (128.0 ± 7.7 ) (P <0.01). For non-stimulated SMCs, MPA directly and significantly inhibited the proliferation of SMCs (29.9 ± 3.5)% at high concentration (10μg · mL-1) At all experimental concentrations, MPA significantly inhibited collagen I product in a dose-dependent manner with a maximum inhibitory rate of (81.0 ± 4.8)%. For ECs-conditioned medium, either MPA The effect of ECs, ECs or conditioned medium preparation after adding MPA to the culture medium, MPA significantly inhibited the proliferation of SMCs and collagen Ⅰ products. Conclusion MPA can inhibit the proliferation of human vascular SMCs to a certain extent and inhibit the collagen Ⅰ products of SMCs.