红花苷对模拟微重力下细胞迁移的作用

Effect of Carthamin on the Cell Migration Induced by Modeled Microgravity

目的观察红花苷修复模拟微重力下人脐静脉内皮细胞(HUVECs)细胞迁移抑制的作用并探讨其机制。方法 MTT比色法选出红花苷干预HUVECs的最佳浓度;模拟失重-2D回转仪模拟失重培养HUVECs,分为正常重力对照组、模拟微重力组、红花苷微重力组。采用透膜侵袭小室模型检测细胞迁移;激光共聚焦显微镜观察红花苷干预后细胞骨架纤维状肌动蛋白(F-actin)的形态及对黏着斑(FAs)的影响。结果与正常重力对照组比较,模拟微重力组细胞迁移数下降(P<0.01);红花苷微重力组较模拟微重力组细胞迁移数增加(P<0.05)。正常重力对照组的F-actin呈现明显极化、微丝集结成束,模拟微重力可导致F-actin骨架重构,极化下降并呈现弥散特点;而红花苷微重力组的F-actin排列有序,伪足较模拟微重力组增多,方向性显著增强。模拟微重力组较正常重力对照组FAs数量(VN)、面积(VA)及FAs到细胞边缘距离(DS)显著降低(P<0.05);红花苷微重力组较模拟微重力组VN、VA、DS显著升高(P<0.05)。结论红花苷可改善模拟微重力作用导致的HUVECs迁移抑制,修复模拟微重力作用导致的HUVECs骨架F-actin的重构,改善FAs的生成及定位。

Objective To observe the repairing effect of Carthamin on the inhibition of cell migration induced by modeled microgravity（MMG）and its mechanism.Methods The proper concentration of Carthamin was got with MTT assay.The human umbilical vein endothelial cells（HUVECs）were cultured and were divided into the normal gravity（NG）control group, MMG group and MMG＋Carthamin group.Transwell was used to detect cell migration.Confocal microscopy and image analysis were used to analyze the modifications of the cytokeratin network and FAs structure.Results Comparing with the NG group,the number of cell migration was significantly decreased in the MMG group （P0.01）.Comparing with the MMG group,the number of cell migration was significantly increased in the MMG＋Carthamin group（P0.05）.In the NG group, F-actin was obviously polarized and microfilaments were assembled into a bundle.The cytoskeleton of HUVECs was refined, F-actin polarization was significantly decreased and microfilaments staining showed diffuse in the MMG group.Comparing with the MMG group,the F-actin of HUVECs was orderly arranged,pseudopodium was increased and the directivity was enhanced in the MMG＋Carthamin group.Comparing with the NG group,the number of FAs（VN）and area（VA）and distance（DS） to cell edge in the MMG group were significantly decreased（P0.05）.Comparing with the MMG group,the VN,VA and DS were significantly increased in the MMG＋Carthamin group（P0.05）.Conclusion Carthamin could repair the blocked HUVECs migration induced by MMG and it is associated with refinement of F-actin and FAs structure.