Zhang Hai-Hong, Chen Yu-Cheng, Liang Ling, Zeng Zhi
Department of Cardiology, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Mar;39(2):188-92.
To study the influence of Tanshinone II A on the proliferation and migration of rabbit vascular smooth muscle cell in vitro and to primarily probe into the mechanism of how Tanshinone II A works.
Rabbit aortic artery was chosen for primary culture of vascular smooth muscle cells (VSMC), which were used and divided into control and Tanshinone II A groups with different drug concentrations. After cells, according to the protocol, incubated with drug for 24 h, 48 h or 72 h respectively, MTT assay was used to study the effect of Tanshinone II A on the VSMC proliferation; sterile scraper and contrast phase microscope were used to measure the distance of VSMC migration; analyses of cell cycle and DNA content were performed by flow cytometry. Then after TRITC-phalloidin labeling for cytoskeleton filament (F-actin), the influence of Tanshinone II A on the cytoskeleton was revealed by laser confocal microscope.
(1) At the same time point, Tanshinone II A inhibited the proliferation and migration of rabbit VSMCs in vitro significantly in a dose-dependent manner; (2) After 24 h, compared with control group Tanshinone II A increased the ratio of rabbit VSMCs in G0/G1 phase, and decreased DNA content significantly in a dose-dependent manner; (3) Tanshinone II A influenced rabbit VSMCs cytoskeleton filament, including relatively short filaments arranged within nonpolarized cells and no lamellipodial protrusions, while in control group formation of lamellipodial protrusions and actin filaments parallel to long axis were seen. CONCLUSION; Tanshinone II A can significantly inhibit the proliferation and migration of rabbit VSMCs in vitro, which may result from arresting rabbit VSMCs through the check point of cell cycle, accumulating them in G0/G1 phase, inducing cell apoptosis and influencing the cytoskeleton filament.
研究丹参酮ⅡA对兔血管平滑肌细胞体外增殖和迁移的影响,并初步探讨其作用机制。
选取兔主动脉进行血管平滑肌细胞(VSMC)原代培养,将其分为对照组和不同药物浓度的丹参酮ⅡA组。细胞按方案分别与药物孵育24 h、48 h或72 h后,采用MTT法研究丹参酮ⅡA对VSMC增殖的影响;用无菌刮棒和相差显微镜测量VSMC迁移距离;通过流式细胞术分析细胞周期和DNA含量。然后用TRITC-鬼笔环肽标记细胞骨架丝(F-肌动蛋白),用激光共聚焦显微镜观察丹参酮ⅡA对细胞骨架的影响。
(1)在同一时间点,丹参酮ⅡA显著抑制兔VSMC体外增殖和迁移,呈剂量依赖性;(2)24 h后,与对照组相比,丹参酮ⅡA增加了兔VSMC在G0/G1期的比例,并呈剂量依赖性显著降低DNA含量;(3)丹参酮ⅡA影响兔VSMC细胞骨架丝,包括非极化细胞内排列相对较短的丝,无片状伪足突出,而对照组可见片状伪足形成和与长轴平行的肌动蛋白丝。结论:丹参酮ⅡA可显著抑制兔VSMC体外增殖和迁移,其机制可能是通过细胞周期检查点阻滞兔VSMC,使其积聚在G0/G1期,诱导细胞凋亡并影响细胞骨架丝。