槲皮素对模拟微重力条件下体外培养大鼠心肌细胞骨架的影响  

Effect of quercetin on the cytoskeleton of rat cardiac myocytes cultured in vitro trader simulated microgravity

作　　者：熊江辉[1] 李莹辉[1] 聂捷琳[1] 丁柏[1] 张晓铀[1] 黄增明[1] 毕蕾[1] 

机构地区：[1]航天医学工程研究所细胞与分子生物学实验室,北京100094

出　　处：《中国生物学文摘》2005年第7期87-91,共5页Chinese Biological Abstracts

基　　金：国家高技术研究发展计划资助项目（No.863-2-2-2-4）

摘　　要：采用细胞免疫双荧光染色观察离体培养的大鼠心肌细胞微丝和微管分布，探讨模拟微重力条件下槲皮素对心肌细胞骨架分布的影响。结果表明：模拟微重力条件下心肌细胞微丝、微管在近胞核区的分布增多：模拟微重力处理的同时加入槲皮素，则使近胞核处微丝、微管分布明显减少，微丝束的粗细与对照组无异。提示模拟微重力可显著影响心肌细胞微丝、微管的分布，槲皮素可对抗该效应而发挥其心肌细胞保护作用。In order to explore effect of quercetin on cytoskeleton distribution in cardiac mycytes under simulated microgravity, we observed the distribution of actins and tubulins, which are the constituentproteins of cardiac myocyte microfilaments and microtubules respectively, using immuuocytochemistry and fluorescence microscope. Enriched cultures of cardiac myocytes were obtained from 2-day-old Wistar rats by stepwise trypsin dissociation, the dispersed cells were incu- bated in cell culture coverslips fixed in cell culture flasks. Cells were divided into three groups： a simulated microgravity group （flasks were set up in a cliuostat and rotated at 30 r/min for 48 hours in a 37℃ incubator） , a control group （cultured in stationary flasks） and a quercetin group （cells were inculated in 100 ug/ml quercetln under cliuorotatlon）. The coverslips was washed in PBS, and fixed with 3. 7 % paraformaldehyde for 30 minutes. After auother wash in PBS and blocking in PBS containing 5 % uormal sheep serum for 30 minutes, the coverslips were incubated with a 1 50 dilution of mouse anti rat α-tubulin mouoantibody for 90 minutes. The coverslips were rinsed with PBS and then incubated with rabzit anti-mouse IgG conjugated with FITC for 150 minutes at room temperature. A wash of PBS was performed and the coverslips were incubated with Texas Red-X phalloidin （1 500） for 120 rain, then the coverslips were rinsed, mounted in crystal mount medium with mowiol andphotographed using laser scanning confocal microscope. Microfilaments and microtubules were stained with green and red fluorescence respectively when observed under a confocal microscope. Compared to the control group, the microfilaments and microtubules of rat cardiac rnyocytes cultured in vitro under simulated microgravity were more heavily concentrated near the nucleus. Treatment with quercetin could partly inhibit the effect of simulated microgravity on the redistribution of microfilaments and microtubules and there was little difference in the thick ness of m

关 键 词：细胞骨架 槲皮素 模拟微重力 体外培养心肌细胞 微重力条件 心肌细胞 体外培养 大鼠 微重力处理 离体培养 

分 类 号：Q693[生物学—生物物理学] TQ511[化学工程]