Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
Pharm Biol. 2013 Sep;51(9):1150-7. doi: 10.3109/13880209.2013.782051. Epub 2013 Jun 12.
Andrographolide, extracted from the leaves of Andrographis paniculata (Burm. f.) Nees (Acanthaceae), is a labdane diterpene lactone. It is widely reported to possess anti-inflammatory and antitumorigenic activities. Cerebral endothelial cells (CECs) play a crucial role in supporting the integrity and the function of the blood-brain barrier (BBB). However, no data are available concerning the effects of andrographolide in CECs. The aim of this study was to examine the detailed mechanisms of andrographolide on CECs.
This study investigated a novel bioactivity of andrographolide on cerebral ischemia/reperfusion-induced brain injury.
CECs were treated with andrographolide (20-100 µΜ) for the indicated times (0-24 h). After the reactions, cell survival rate and cytotoxicity were tested by the MTT assay and the lactate dehydrogenase (LDH) test, respectively. Western blotting was used to detect caspase-3 expression. In addition, analysis of cell cycle and apoptosis using PI staining and annexin V-FITC/PI labeling, respectively, was performed by flow cytometry. We also investigated the effect of andrographolide on middle cerebral artery occlusion (MCAO)/reperfusion-induced brain injury in a rat model.
In the present study, we found that andrographolide (50-100 µΜ) markedly inhibited CEC growth according to an MTT assay and caused CEC damage according to a LDH test. Our data also revealed that andrographolide (50 µM) induced CEC apoptosis and caspase-3 activation as respectively detected by PI/annexin-V double staining and western blotting. Moreover, andrographolide arrested the CEC cell cycle at the G0/G1 phase by PI staining. In addition, andrographolide (5 mg/kg) caused deterioration of MCAO/reperfusion-induced brain injury in a rat model.
These data suggest that andrographolide may disrupt BBB integrity, thereby deteriorating MCAO/reperfusion-induced brain injury, which are, in part, associated with its capacity to arrest cell-cycle and induce CEC apoptosis.
穿心莲内酯是从穿心莲（Burm. f.） Nees（爵床科）的叶子中提取的一种贝壳杉烷二萜内酯。它被广泛报道具有抗炎和抗肿瘤活性。脑内皮细胞（CEC）在维持血脑屏障（BBB）的完整性和功能方面起着至关重要的作用。然而，目前尚无关于穿心莲内酯对 CEC 的影响的数据。本研究旨在研究穿心莲内酯对 CEC 的详细作用机制。
本研究探讨了穿心莲内酯在脑缺血/再灌注诱导的脑损伤中的一种新的生物活性。
用穿心莲内酯（20-100μM）处理 CEC 不同时间（0-24 小时）。反应后，通过 MTT 检测法和乳酸脱氢酶（LDH）试验分别检测细胞存活率和细胞毒性。用 Western blot 检测 caspase-3 的表达。此外，通过 PI 染色和 Annexin V-FITC/PI 标记分别用流式细胞术分析细胞周期和细胞凋亡。我们还研究了穿心莲内酯对大鼠大脑中动脉闭塞（MCAO）/再灌注诱导的脑损伤的影响。
在本研究中，我们发现穿心莲内酯（50-100μM）显著抑制 MTT 检测法中的 CEC 生长，并根据 LDH 试验导致 CEC 损伤。我们的数据还表明，穿心莲内酯（50μM）通过 PI/annexin-V 双重染色和 Western blot 分别检测到诱导 CEC 凋亡和 caspase-3 激活。此外，穿心莲内酯通过 PI 染色将 CEC 细胞周期阻滞在 G0/G1 期。此外，穿心莲内酯（5mg/kg）在大鼠模型中加重 MCAO/再灌注诱导的脑损伤。
这些数据表明，穿心莲内酯可能破坏 BBB 的完整性，从而加重 MCAO/再灌注诱导的脑损伤，这在一定程度上与其阻滞细胞周期和诱导 CEC 凋亡的能力有关。
