模拟微重力对大鼠嗜铬细胞瘤细胞形态及其生长的影响 Effects of Simulated Microgravity on the Morphology and Growth of PC12 Cells.pdfVIP

第19 卷 第2 期 航天医学与医学工程 VOI. 19 NO. 2 2006 年 4 月 Space Medicine MedicaI Engineering Apr. 2006 ·学术论著 · 模拟微重力对大鼠嗜铬细胞瘤细胞形态及其生长的影响＊ 杨唐斌，曲丽娜，钟 萍，元艳宏，肇 海 （中国航天员科研训练中心，北京 100094 ） 摘要：目的研究回转是否影响PC12 细胞内蛋白质的羰基化和硝基化水平，进而影响细胞骨架和形态； 细胞内活性氮水平升高是否会影响PC12 细胞的增殖分化。方法观察并测定了静置对照组和回转组细 胞的细胞形态和细胞密度的变化，采用免疫荧光标记和RT- PCR 技术研究了回转对细胞骨架蛋白Actin 羰基化和Tubulin 硝基化以及nNOS 和iNOS 的mRNA 和蛋白表达的影响；并利用流式细胞术检测了 细胞周期的改变，初步探讨了这些改变和一氧化氮合酶之间的关系。结果 细胞骨架蛋白发生硝基化和 羰基化的改变，但回转组细胞骨架形态变化不明显，PC12 细胞的微丝回转后明显伸长；回转后，细胞密 度显著增加，细胞周期中DNA 合成的S 期明显缩短。结论 结果表明水平回转模拟微重力并不改变 PC12 细胞形态，相反还有利于细胞的生长，其机制可能与一氧化氮合酶的表达上调有关。 关键词：失重模拟；水平式回转器；硝基酪氨酸；蛋白羰基；细胞骨架；荧光标记 中图分类号： R854 文献标识码：A 文章编号：1002-0837 （2006 ）02-0079-06 Eff ects of Simulated Microgravity on the Morp hology and Growth of PC12 Cells. YANG Tang- bin，OU Li- na ，ZHONG Ping ，Y UAN Yan- hong ，ZHAO Hai. Space Medicine Medical Engineering ，2006 ，19 （2 ）： 79 ~ 84 Abstract ：Objective To observe whether the increase of oxidative stress in PC12 cells could influ- ence the levels of protein carbonyls and nitrotyrosine and alter the cytoskeleton and cell morphology under clinostat condition，and whether the increase of content of nitrate/ nitrite in cell culture medi- um could influence the cell proliferation and differentiation. Method Cell morphology ，carbonylated actin and nitrotyrosinated tubulin，and mRNA and protein express of nNOS and iNOS were observed and determined with immunofluorescence and RT- PCR technology in clinostat rotated and control static groups. At the same time ，cell density was measured and cell cycles were detected with flow cytometry . The relationship between all these changes and NOS were also analyzed. Result The levels of carbonylated and nitrotyrosinated cytoskeleton protein were alt