模拟微重力效应对成骨细胞纺锤体结构的影响

发布时间：2018-06-26 04:08

本文选题：模拟微重应力效 + 成骨细胞　；参考：《哈尔滨工业大学》2011年硕士论文

【摘要】：航天事业的发展推动航天医学、空间细胞生物学的兴起与进步,而微重力对细胞各方面的影响则成为研究的重点。微重力对细胞形态及细胞骨架、周期、凋亡等各项生理指标发挥重要作用、意义深远。本课题组前期研究表明,模拟微重力效应使人骨肉瘤细胞微管细胞骨架和纺锤体结构改变,细胞周期阻滞,以及纺锤体检验点蛋白表达改变。为了进一步探讨模拟微重力效应对成骨细胞的影响,本实验通过回转模拟微重力效应,检测不同时间点下其对人成骨细胞系hFOB1.19和大鼠原代成骨细胞包括微管骨架、纺锤体结构、细胞周期、细胞凋亡的影响情况。本研究利用回转器模拟微重力效应,分别培养hFOB1.19细胞及原代成骨细胞24、48、72、96小时。通过免疫荧光技术标记α-tubulin,激光共聚焦显微镜观察发现模拟微重力效应48小时后hFOB1.19细胞纺锤体结构异常,出现三极、四极纺锤体,模拟微重力效应72、96小时后纺锤体结构异常加剧,出现五极、六极纺锤体,4个时间点内多极纺锤体形成率分别为3.87%、10.08%、11.94%、12.16%;而原代成骨细胞在模拟微重力效应96小时后才观察到三极纺锤体,且多极纺锤体形成率维持在较低水平。流式细胞术检测细胞周期,结果发现模拟微重力效应下hFOB1.19细胞S期显著增多;而原代成骨细胞在模拟微重力效应72小时G2/M期明显增多。模拟微重力效应对2种成骨细胞细胞凋亡的影响均表现为细胞早期凋亡与晚期死亡比例明显增大。实验结果显示,模拟微重力效应导致成骨细胞微管结构紊乱、有序性下降,并导致多极纺锤体的增多,使成骨细胞无法正常分裂,因此造成了细胞周期阻滞。另外,伴随模拟微重力效应时间的延长,成骨细胞微管与纺锤体结构的损伤程度在不断地加剧,证明细胞并未成功得以修复,因而凋亡的细胞也就明显增加。
[Abstract]:The development of space industry promotes space medicine, the rise and progress of space cell biology, and the influence of microgravity on all aspects of cells has become the focus of research. Microgravity plays an important role in cell morphology, cytoskeleton, cycle, apoptosis and other physiological indexes.
The previous study showed that simulated microgravity effect made the microtubule of human osteosarcoma cell microtubule cytoskeleton and spindle structure change, cell cycle block, and the change of protein expression of spindle checkpoint. In order to further explore the effect of simulated microgravity effect on osteoblasts, this experiment was not detected by rotary simulation microgravity effect. At the same time, its effects on human osteoblast cell line hFOB1.19 and rat primary osteoblasts including microtubule skeleton, spindle structure, cell cycle and apoptosis were observed.
In this study, hFOB1.19 cells and primary osteoblasts were cultured for 24,48,72,96 hours by using a gyroscope to simulate the effect of microgravity. By labeling alpha -tubulin by immunofluorescence technique, the spindle structure of hFOB1.19 cells was found to be abnormal after 48 hours of simulated microgravity, and three pole, quadrupole spindle, and simulated microweight appeared. When the force effect was 72,96 hours, the spindle structure was abnormal. The formation rate of the multipolar spindle was 3.87%, 10.08%, 11.94% and 12.16%, respectively, and the formation rate of the primary osteoblast was 3.87%, 11.94% and 12.16%, respectively, and the formation rate of the multipolar spindle was maintained at a lower level after the simulated microgravity effect for 96 hours. Cell cycle was detected by cell operation. The results showed that the S phase of hFOB1.19 cells increased significantly under simulated microgravity, while the primary osteoblasts were significantly increased in the 72 hour G2/M phase of simulated microgravity effect. The effect of simulated microgravity effect on the apoptosis of 2 osteoblast cells showed that the proportion of early apoptosis and late death increased significantly.
The results showed that the simulated microgravity effect resulted in the disorder of the microtubule of osteoblasts, the decreasing of the order, the increase of the multipolar spindle and the failure of the osteoblasts to split normally, thus causing cell cycle arrest. In addition, the damage degree of the microtubule and spindle structure of osteoblasts was associated with the prolonged duration of the simulated microgravity effect. Increasing evidence shows that cells have not been successfully repaired, and the apoptotic cells have increased significantly.
【学位授予单位】：哈尔滨工业大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R85

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