Zhao Qing, Huang Hai-xiao, Jin Chun-hua
Key Laboratory for Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2003 Apr;23(4):364-6.
To observe the effect of lipopolysaccharide (LPS) on actin cytoskeleton of rat cardiac myocytes and the intervention effect of polydatin against this effect.
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.
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.
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.
观察脂多糖(LPS)对大鼠心肌细胞肌动蛋白细胞骨架的影响以及虎杖苷对此影响的干预作用。
从新生SD大鼠(3日龄)分离心肌细胞并进行体外培养,然后将其分为对照组(用D - 汉克氏溶液处理30分钟)、虎杖苷组(用0.2 mmol/L虎杖苷处理10分钟)、LPS组(用100 ng/ml LPS刺激30分钟)和LPS/虎杖苷组(用100 ng/ml LPS刺激30分钟,随后用0.2 mmol/L虎杖苷孵育10分钟)。处理完成后,通过免疫荧光染色分析细胞中的心肌F - 肌动蛋白。
对照组中,F - 肌动蛋白定位于心肌细胞的皮质,细胞内充满了组织成网状结构的F - 肌动蛋白。LPS刺激后,皮质中F - 肌动蛋白的染色变淡甚至不可见,细胞中观察到应力纤维形成,经10分钟虎杖苷处理后应力纤维消失,F - 肌动蛋白恢复正常排列。对照组和虎杖苷组之间未发现明显差异。
LPS可能直接诱导应力纤维形成,从而对大鼠心肌细胞造成损伤,虎杖苷可通过参与F - 肌动蛋白组织的机制使其恢复。