持续性压力对PC12细胞增殖和细胞骨架蛋白表达的研究  

Effects of continuous compression on the proliferation of PC12 cells and the expression of cytoskeletal protein

作　　者：方钧[1] 叶正旭[1] 王宇清[1] 杜俊杰[1] 罗卓荆[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,西安710032

出　　处：《武警医学》2010年第5期383-387,共5页Medical Journal of the Chinese People's Armed Police Force

基　　金：军队"十一五"科技攻关资助计划项目(06G097)

摘　　要：目的研究持续性压力对PC12细胞增殖和细胞骨架蛋白表达的影响。方法利用自制压力可控性细胞损伤装置,对PC12细胞分别施加0.1、0.3、0.5mPa的持续压力,且在不同压力值作用下分别持续加压10、30、60min,应用MTT法和免疫组化染色法分析不同压力值及持续时间对PC12细胞增殖和细胞骨架蛋白表达的影响。结果正常PC12细胞呈上皮样细胞生长,表现出神经细胞特性。当加压损伤后,PC12细胞出现凋亡、坏死现象。对照组MTT结果显示,PC12细胞的OD值随观察期延长而逐渐升高;而加压组中,当压力≥0.3mPa、持续时间≥30min后的细胞增殖显著受抑(P<0.05)。β-tubu-linⅢ免疫组化染色结果显示0.5mPa压力作用10min后,细胞骨架蛋白β-tubulinⅢ逐渐断裂降解,且降解程度随持续作用时间的延长而增强。结论在急性压迫性脊髓损伤中,早期神经细胞增殖受限和β-tubulinⅢ蛋白降解,可随压力值和持续时间增加而增强,这种现象可能参与了ASCI后机械信号转变为生物学信号的过程。Objectives To investigate the effect of continuous compression on the proliferation and the expression of cytoskcletal protein in cultured PC12 cells in vitro. Methods A self - made hydrostatic compressive device was used to apply 0.1,0.3, 0. 5 mPa static continuous stress on cultured PC12 cells, which was also applied at 10, 30 or 60min under different pressure. Then the MTY and immunohistochemical methods were used to study the effect of different continuously compressive pressure of different duration on the cellular proliferation and expression of cytoskeletal protein in cultured PC12 cells in vitro. Results The morphology of cultured PCI2 cells, which resembled the Epithelioid cells, showed the characteristics of neural cells in culture. At a higher pressure, the PC12 cells showed apoptosis and necrosis. The MTT results showed that the values of absorbance in control group gradually increased over time. The cellar proliferation was significantly inhibited after 30 min and at a pressure ≥0.3 mPa （ P 〈 0.05 ）. The immunohistochemical results of β -tubulinm showed the cytoskeletal protein degradation after 0.5 mPa application of 10 min and enhanced degradation with the extension of duration. Conclusions In acute compressive spinal cord injury, the degree of neural cell proliferation and β - tubulin III protein degradation may be positively correlated with the pressure value and duration, which might be involved in the transformation of the mechanical signal into a biological signal after ASCI.

关 键 词：PC12细胞 细胞增殖 Β-微管蛋白 压力 

分 类 号：R681.54[医药卫生—骨科学]