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 [动物学报49(1) :98～ 10 3 ,2 0 0 3]。
In order to explore effect of quercetin on cytoskeleton distribution in cardiac myocytes under simulated microgravity, we observed the distribution of actins and tubulins, which are the constituent proteins of cardiac myocyte microfilaments and microtubules respectively, using immunocytochemistry and fluorescence microscope.Enriched cultures of cardiac myocytes were obtained from 2-day-old Wistar rats by stepwise trypsin dissociation, the dispersed cells were incubated 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 clinostat 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 quercetin under clinorotation).The coverslips was washed in PBS, and fixed with 3.7% paraformaldehyde for 30 minutes.After another wash in PBS and blocking in PBS containing 5% normal sheep serum for 30 minutes, the coverslips were incubated with a 1∶50 dilution of mouse anti rat α-tubulin monoantibody for 90 minutes.The coverslips were rinsed with PBS and then incubated with rabbit 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 min, then the coverslips were rinsed, mounted in crystal mount medium with mowiol and photographed 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 myocytes 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 thickness of microfilaments between the quercetin and control group.These results suggest that treatment with simulated microgravity results in the redistribution of microfilaments and microtubules of rat cardiac myocytes cultured in vitro.Quercetin shows its protective effect via partial inhibition of the cytoskeleton redistribution in cardiac myocytes induced by simulated microgravity [Acta Zoologica Sinica 49(1):98-103, 2003].
出处
《动物学报》
SCIE
CAS
CSCD
北大核心
2003年第1期98-103,共6页
ACTA ZOOLOGICA SINICA
基金
国家高技术研究发展计划资助项目 (No. 86- 3 -2 -2 - 2 - 4)~~
