钙调蛋白拮抗剂W-7对人Tenon囊成纤维细胞的抑制实验

目的观察钙调蛋白拮抗剂W-7对培养的人Tenon囊成纤维细胞的抑制作用。方法1)人Tenon囊成纤维细胞离体培养及生长抑制实验:取对数生长期的Tenon囊成纤维细胞接种于96孔培养板中,分别加入不同浓度的W-7及丝裂霉素C(MMC),培养不同时间,用MTT法检测细胞增生抑制情况。2)药物对人Tenon囊成纤维细胞形态及线粒体代谢的影响:将人Tenon囊成纤维细胞接种在96孔培养板中,同时加入不同浓度的W-7及MMC,分别培养不同时间,观察细胞形态变化;同时加入MTT,观察细胞线粒体代谢变化。结果1)W-7依剂量及时间依赖方式明显抑制离体培养的人Tenon囊成纤维细胞增生。2)MTT显示W-7作用的细胞线粒体代谢存在,而MMC作用后细胞死亡,线粒体代谢消失。结论W-7可抑制人Tenon囊成纤维细胞增生。其对细胞线粒体代谢的影响较MMC小,不引起细胞膜完整性的破坏,可减轻炎症反应,是具有良好应用前景的抗斑痕药物。

Effect of Calmodulin Antagonist W-7 on the Proliferation in Vitro Cultured Human Tenon's Capsule Fibroblasts

Objective To evaluate the effects of calmodulin antagonist W-7 on the proliferation of in vitro cultured human Tenon's capsule fibroblast cells for finding new antiproliferative drugs applied for the glaucoma filtration surgery. Methods Human Tenon's capsule fibroblast cells were cultured in DMEM medium containing 10% fetal bovine serum at 37 ℃,5% CO₂. For growth inhibition assays, cells lie in logarithm growth period were seeded into 96 well plate, and treated with W-7 and MMC, the inhibitory effect was determined by MTT(tetrazolium bromide) method. The mitochondrial metabolic ability of the drugs treated cells was also examined by MTT assays. Results W-7 caused significant inhibition effect of fibroblast proliferation in dose and time dependent manner, which were perhaps the causes of cell apoptosis in human Tenon's capsule fibroblast cells. The MTT assay showed that W-7 treated cells had mitochondrial metabolic ability while MMC treated cells lost this ability. While the cell membrane integrity of W-7 treated cells was superior to MMC treated cells, and it may cause slight inflammatory reaction. Conclusion There is a good prospect of W-7 for clinical application.