發(fā)布時(shí)間：2018-06-26 04:08

  本文選題：模擬微重應(yīng)力效 + 成骨細(xì)胞�。� 參考：《哈爾濱工業(yè)大學(xué)》2011年碩士論文

【摘要】：航天事業(yè)的發(fā)展推動(dòng)航天醫(yī)學(xué)、空間細(xì)胞生物學(xué)的興起與進(jìn)步,而微重力對(duì)細(xì)胞各方面的影響則成為研究的重點(diǎn)。微重力對(duì)細(xì)胞形態(tài)及細(xì)胞骨架、周期、凋亡等各項(xiàng)生理指標(biāo)發(fā)揮重要作用、意義深遠(yuǎn)。 本課題組前期研究表明,模擬微重力效應(yīng)使人骨肉瘤細(xì)胞微管細(xì)胞骨架和紡錘體結(jié)構(gòu)改變,細(xì)胞周期阻滯,以及紡錘體檢驗(yàn)點(diǎn)蛋白表達(dá)改變。為了進(jìn)一步探討模擬微重力效應(yīng)對(duì)成骨細(xì)胞的影響,本實(shí)驗(yàn)通過(guò)回轉(zhuǎn)模擬微重力效應(yīng),檢測(cè)不同時(shí)間點(diǎn)下其對(duì)人成骨細(xì)胞系hFOB1.19和大鼠原代成骨細(xì)胞包括微管骨架、紡錘體結(jié)構(gòu)、細(xì)胞周期、細(xì)胞凋亡的影響情況。 本研究利用回轉(zhuǎn)器模擬微重力效應(yīng),分別培養(yǎng)hFOB1.19細(xì)胞及原代成骨細(xì)胞24、48、72、96小時(shí)。通過(guò)免疫熒光技術(shù)標(biāo)記α-tubulin,激光共聚焦顯微鏡觀察發(fā)現(xiàn)模擬微重力效應(yīng)48小時(shí)后hFOB1.19細(xì)胞紡錘體結(jié)構(gòu)異常,出現(xiàn)三極、四極紡錘體,模擬微重力效應(yīng)72、96小時(shí)后紡錘體結(jié)構(gòu)異常加劇,出現(xiàn)五極、六極紡錘體,4個(gè)時(shí)間點(diǎn)內(nèi)多極紡錘體形成率分別為3.87%、10.08%、11.94%、12.16%;而原代成骨細(xì)胞在模擬微重力效應(yīng)96小時(shí)后才觀察到三極紡錘體,且多極紡錘體形成率維持在較低水平。流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期,結(jié)果發(fā)現(xiàn)模擬微重力效應(yīng)下hFOB1.19細(xì)胞S期顯著增多;而原代成骨細(xì)胞在模擬微重力效應(yīng)72小時(shí)G2/M期明顯增多。模擬微重力效應(yīng)對(duì)2種成骨細(xì)胞細(xì)胞凋亡的影響均表現(xiàn)為細(xì)胞早期凋亡與晚期死亡比例明顯增大。 實(shí)驗(yàn)結(jié)果顯示,模擬微重力效應(yīng)導(dǎo)致成骨細(xì)胞微管結(jié)構(gòu)紊亂、有序性下降,并導(dǎo)致多極紡錘體的增多,使成骨細(xì)胞無(wú)法正常分裂,因此造成了細(xì)胞周期阻滯。另外,伴隨模擬微重力效應(yīng)時(shí)間的延長(zhǎng),成骨細(xì)胞微管與紡錘體結(jié)構(gòu)的損傷程度在不斷地加劇,證明細(xì)胞并未成功得以修復(fù),因而凋亡的細(xì)胞也就明顯增加。
[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.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R85

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