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机构地区:[1]第一军医大学全军休克微循环重点实验室,广东广州510515 [2]第一军医大学实验管理中心,广东广州510515
出 处:《第一军医大学学报》2003年第4期364-366,共3页Journal of First Military Medical University
摘 要:目的观察脂多糖(LPS)对心肌细胞微丝蛋白的形态学影响以及中药虎杖甙(PD)的防治效果。方法体外乳鼠心肌细胞培养,随机分为4组:正常对照组(D'-Hanks 作用于心肌细胞30 min)、PD组(0.2 mmol/L作用于心肌细胞10 min)、LPS组(100 ng/ml刺激30 min)和LPS+PD组(100 ng/ml LPS 30 min, 0.2 mmol/L PD治疗10 min)。用免疫荧光技术标记纤维状肌动蛋白(F-actin)。结果正常对照组心肌细胞皮质部分有较多纤维状肌动蛋白分布,细胞内纤维状肌动蛋白呈网状布满于整个细胞;LPS组细胞皮质部分染色较弱甚至消失,心肌细胞应力纤维(stress fiber)形成;PD(0.2 mmol/L)处理10 min后,心肌细胞应力纤维消失,形态趋于正常组细胞;而PD组和正常对照组细胞形态无明显差异。结论LPS可能直接导致心肌细胞应力纤维的形成,PD对LPS导致的心肌细胞损伤有保护作用。Objective To observe the effect of lipopolysaccharide (LPS) on actin cytoskeleton of rat cardiac myocytes and the intervention effect of polydatin against this effect. Methods Rat cardiac myocytes were isolated from newborn SD rats (3 days old) and cultured in vitro, which were then divided into control group (treated with D-Hank's solution for 30 min), polydatin group (with 0.2 mmol/L polydatin treatment for 10 min), LPS group (with 100 ng/ml LPS stimulation for 30 min), and LPS/polydatin group (with 100 ng/ml LPS stimulation for 30 min followed by incubation with 0.2 mmol/L polydatin for 10 min). When the treatments were completed, the cells were analyzed for myocardial F-actin by immunofluorescent staining. Results In the control group, F-actin was localized in the cortex of cardiac myocytes and the cells were filled with F-actin organized into reticular structures. After LPS stimulation, the staining for F-actin was faint or even invisible in the cortex, with the formation of stress fibers observed in the cells, which disappeared upon the 10-min polydatin treatment and the F-actin resumed normal arrangement. No obvious difference was found between the control and polydatin groups. Conclusion LPS may directly induce stress fiber formation, therefore cause damages to rat cardiac myocytes, which can be reverted by polydatin through the mechanism of participating in the F-actin organization.
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