模拟微重力导致人脐静脉内皮细胞微管骨架重构及增殖的影响

Effect of Modeled Microgravity on Cytoskeleton Architecture and Proliferation of HUVECs

目的:观察模拟微重力(MMG)致人脐静脉内皮细胞(HUVECs)微管骨架结构的改变,并对MMG作用后细胞的增殖等功能进行评价。方法:酶消化法原代培养HUVECs,随机分为2D-clinostat干预的MMG培养组与NG对照培养组,均培养48 h;倒置显微镜下观察细胞形态,细胞计数及流式细胞术分析细胞增殖、凋亡及细胞周期变化。结果:所培养的HUVECs细胞并经流式细胞术鉴定证实;MMG干预48 h使大量的微管蛋白发生解聚,微管的网状结构已经模糊不见,随之代替的是崩解的微管小聚体;MMG可使HUVECs增殖明显抑制,细胞周期抑制于G2/M期(G2/M:MMG,27.6%;NG,18.1%;P<0.05),然而细胞并没有发生明显凋亡。结论:MMG可显著影响HUVECs的形态与细胞骨架结构并抑制其增殖功能,HUVECs的增殖抑制可能与紊乱的微管结构密切相关。

Objective： To investigate the cytoskeleton, proliferation function of HUVECs under MMG. Method： Isolation and culture of HUVECs by enzyme digestion, and cells were identified by flow cytometry （FCM）. The stimulated weightlessness was mimic- ked by 2D- clinostat. HUVECs were exposed to clinorotation for 48 h at 30 rpm. Cell proliferation, apoptosis and cell cycle were analyz- ed by cell counting and FCM. The change of the microtubules network and chromatin structure were observed by Confocal microscopy. Results： The expression of CD31 and Factor- Vm were positive in our primary cultured HUVECs; Instead of long, strongly labeled microtubules radiating throughout the cytoplasm, only a few filaments could be distinguished against the strong background. This more or less diffuse labeling could correspond to either labeled free tubulin subunits or numerous but very short microtubules; HUVECs prolif- eration was greatly decreased in MMG while no apoptosis was detected and cell Cycling was mainly blocked in G2/M phase （MMG, 27.6%; NG, 18.1%; P〈0.05）. Conclusions： MMG could affect cellular morphology and proliferation. And the decreased cell proliferation is greatly associated with architecture of microtubules.