目的:探讨生理电场干预对大鼠主动脉血管平滑肌细胞形态和细胞骨架蛋白F-actin表达的影响。 方法:实验于2002-07/2003-12在解放军第三军医大学西南医院中心实验室进行。取清洁级Wistar大鼠47只,颈离断法处死大鼠,体外培养其主动脉血管平滑肌细胞。①分别给予0,50,100,150,200,250mV/mm生理电场干预,显微成像及图像分析技术记录分析血管平滑肌细胞形态变化。②150mV/mm生理电场干预的血管平滑肌细胞在无生理电场(0)和生理电场作用5,10,30min、1,3,6h各时间点,用激光共聚焦显微镜检测细胞骨架蛋白F-actin的变化。 结果:①血管平滑肌细胞在100~250mV/mm生理电场作用下显示明显的趋化反应,部分细胞的板状突起向阴极面伸展。②激光共聚焦显微镜显示150mV/mm生理电场作用5min,细胞内F-actin表达增加,30~60min达到高峰,以后仍然高于无电场干预的血管平滑肌细胞;F-actin逐渐呈平行排列,形成整齐的应力纤维丝。结论:生理电场干预下血管平滑肌细胞细胞膜向阴极伸展,血管平滑肌细胞F-actin合成和重新分布,发生结构改变,可能参与血管平滑肌细胞定向迁移的调控。
AIM: To investigate the effects of intervention of physiological electric field (EF) on morphous of vascular smooth muscle cells (VSMCs) of arteriae aorta (AO) and expression of cytoskeletal protein F-actin in rats. METHODS: The experiment was conducted in the Central Laboratory of Southwest Hospital, Third Military Hospital of Chinese PLA from July 2002 to December 2003. Totally 47 Wistar rats of clean grade were selected. Rats were executed with neck mutilating method; VSMCs of AO were cultured in vitro. ①Intervention of physiological EF were given at 0, 50, 100,150,200,250 mV/mm respectively, morphologic changes of VSMCs were recorded and analyzed with micro-imaging and image analysis method. ②Changes of cytoskeletal protein F-actin in VSMCs with intervention of physiological EF were inspected with laser confocal microscopy at each time-point such as in the 5^th minute, 10^th minute, 30^th minute, 1^st hour, 3rd hour and 6^th hour of action in non-physiological EF (0) and physiological field. RESULTS:①There was obvious chemotactic response (CKT) in VSMCs in EF of 100-250 mV/mm, partial cellular buttressed prominences extended toward negative pole. ②Confocal microscopy showed that expression of F-actin in cells increased in the 5^th minute of action in physiological EF at 150 mV/mm, and reached to the peak between 30-60 minutes, it was higher than that in VSMCs without intervention of EF and F-actin were in ordered parallel arrays gradually and formed into concinnous stress fiber. CONCLUSION: Under the intervention of physiological EF, cellular membrane of VSMCs extend towards negative pole. There are synthesis and redistribution of Foactin in VSMCs and structures changes, which may be involved in the regulation of oriented migration of VSMC.