舒林酸代谢产物诱导人血管内皮细胞凋亡的的实验研究
Sulindac derivative-induced apoptosis in a human umbilical vein endothelial cell line ECV304
摘要目的探讨舒林酸硫化物及砜化物体外对血管内皮细胞增殖及凋亡的影响。方法 MTT法测定舒林酸硫化物及砜化物对人脐静脉内皮细胞增殖的作用,流式细胞仪和电镜分别检测其细胞周期、凋亡和超微结构的改变。 结果 MTT显示,舒林酸硫化物可显著抑制血管内皮细胞的增殖,呈剂量依赖效应,IC50为200 μmol/L。流式细胞仪检测结果显示,舒林酸硫化物可改变血管内皮细胞的周期分布,G0-G1期(对照组77.74%±1.58%,舒林酸砜化物组75.63%±2.12%,舒林酸硫化物组46.12%±1.60%),S期(对照组13.64%±1.22%,舒林酸砜化物组16.40%±2.30%,舒林酸硫化物组27.26%±2.08%),G2-M期(对照组8.61%±0.67%,舒林酸砜化物组7.98%±0.49%,舒林酸硫化物组26.62%±3.54%)。与对照组比较,舒林酸硫化物使G0-G1期细胞显著减少(P<0.01),S期增加(P<0.01),G2-M增加(P<0.01)。舒林酸硫化物使细胞凋亡率增加(对照组6.08%±3.39%,舒林酸砜化物组4.81%±2.14%,舒林酸硫化物组51.90%±5.67%), 与对照组比较,差异有极显著意义(P<0.01)。电镜下舒林酸硫化物组可见细胞核染色质浓缩,细胞膜发泡,并出现凋亡小体。而舒林酸砜化物对血管内皮细胞的增殖、细胞周期分布及凋亡无影响。结论 舒林酸的硫化代谢产物能够显著抑制血管内皮细胞的增殖活性,改变细胞的周期分布,使细胞阻滞于G2期,并可能在此期诱导其凋亡。舒林酸砜化物对血管内皮细胞的增殖、细胞周期分布及凋亡无影响。
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abstractsObjective To investigate the effects of sulindac metabolites on proliferation and apoptosis in the human umbilical vein endothelial cell line ECV304 in vitro.Methods The proliferation profile of ECV304 was determined by methyl thiazolyl tetrazolium (MTT) method. Cell cycle distribution, apoptosis and the ultrastructure of ECV304 were detected by flow cytometry (FCM) and electron microscopy, respectively.Results MTT assay showed that the sulfide inhibited the proliferation of ECV304 and its effect was dose-dependent; the IC50 was 200 μmol/L. FCM showed that the sulfide changed cell cycle distribution. The cell cycle distribution was as follows: G1 phase (control group 77.74%±1.58%; sulfone group 75.63%±2.12%; sulfide group 46.12%±1.60%); S phase (control group 13.64%±1.22%; sulfone group 16.40±2.30%; sulfide group 27.26%±2.08%); G2-M phase (control group 8.61%±0.67%; sulfone group 7.98%±0.49%; sulfide group 26.62%±3.54%). The apoptosis rates in the control group, sulfone group and sulfide group were 6.08%±3.39%, 4.81%±2.14% and 51.90%±5.67%, respectively. Sulfide reduced the proportion of G1 phase, increased the proportion of S phase, G2-M phase and the apoptosis rate significantly (P<0.01, vs control). In the sulfide-treated cells, there were nuclear fragmentation and chromosomal condensation, shrinkage of the cell and loss of contact with neighboring cells. Apoptotic bodies were observed. Sulfone showed no effect on cell proliferation, cell cycle distribution or cell morphology.Conclusions Sulfide can significantly reduce the proliferation of ECV304, change the cell cycle distribution and arrest cells in G2-M phase where apoptosis may be induced. Sulfone has no such effects on this cell line.
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