甲状腺素在细胞超微水平对人脐静脉血管内皮细胞增殖及迁移的影响

Effects of thyroxine hormone on proliferation and migration of human umbilical vein endothelial cells at ultrastructure level

目的：观察甲状腺素对体外培养的人脐静脉血管内皮细胞（ HUVECs ）增殖、迁移的影响。方法采用噻唑兰比色法检测各浓度（0、10^-10、10^-9、10^-8、10^-7、10^-6 mmol/L）甲状腺素对HUVECs增殖的影响；细胞划痕实验及Transwell实验检测甲状腺素（0、10^-9、10^-7 mmol/L）对HUVECs迁移的影响；利用原子力显微镜（ AFM）观察甲状腺素（0、10^-9、10^-7 mmol/L）对HUVECs 细胞表面超微结构的影响。结果与0 mmol/L相比，10^-10、10^-9、10^-8、10^-7、10^-6 mmol/L甲状腺素处理的HUVECs的增殖率明显增大，增殖率分别为（11.572±0.68）％、（18.549±7.53）％、（24.183±4.02）％、（38.724±1.98）％、（32.492±0.76）％，并且细胞增殖程度与甲状腺素浓度（10-10～10-7 mmol/L）呈剂量依赖关系（P＜0.01）；10^-9、10^-7 mmol/L甲状腺素处理的HUVECs的迁移距离和迁移数量均较0 mmol/L明显增大（ P＜0.01），迁移距离和迁移数量依处理浓度的由低到高分别为（18.649±2.370）、（58.176±4.163）、（140.354±25.239）μm和（37.673±2.302）、（73.860±2.608）、（117.405±5.128）个；甲状腺素可使HUVECs 表面超微结构发生明显改变，表现为细胞表面粗糙度增加，突起增大及数量增多。结论甲状腺素可以改变HUVECs表面的超微结构，促进血管内皮细胞形成突起及伪足，进一步促进血管内皮细胞的增殖、迁移。

Objective To observe the effects of thyroxine hormone （ TH） on the proliferation and migration of human umbilical vein endothelial cells （HUVECs）.Methods HUVECs were co-cultured with TH.The proliferation rate of HUVECs was examined by MTT assay , while the cell migration ability was assessed by using the modified of wound healing assay and transwell chamber assay .The morphology and ultrastructure observation of HUVECs membrane surface was carried out by atomic force microscope （ AFM） .Results TH significantly induced proliferation of HUVECs in a dose -dependent manner （ P〈0.01 ） .The proliferation rates of HUVECs in each group （ 10 -10 , 10 -9 , 10 -8 10-7 and 10 -6 mmol/L） were （11.572 ±0.68）%, （18.549 ±7.53）%, （24.183 ±4.02）%, （38.724 ±1.98）% and （32.492 ± 0.76）%, respectively.The migration distance of HUVECs and the number of cells passing the transwell chamber were both significantly increased in the groups treated with TH than in control group （0 mmol/L） （P〈0.01）.The migration distances in control, 10-9 mmol/L and 10-7 mmol/L groups were （18.649 ±2.370）, （58.176 ±4.163） and （140.354 ± 25.239）μm, respectively;while the numbers of HUVECs passing the transwell chamber were 37.673 ±2.302, 73.860 ± 2.608 and 117.405 ±5.128, respectively.According to AFM image, HUVECs were more active after the treatment with TH than the control group （0 mmol/L） cells, presenting with more pseudopodium , granules extended the edges of cell and rough surface in cell membrane .Conclusion TH can change the ultrastructure of the HUVECs surface thus to promote cell proliferation and migration .