Hur Kyu Yeon, Seo Hye Jun, Kang Eun Seok, Kim Soo Hyun, Song Seungjeong, Kim Eun Hee, Lim Soyeon, Choi Chulhee, Heo Ji Hoe, Hwang Ki Chul, Ahn Chul Woo, Cha Bong Soo, Jung Mankil, Lee Hyun Chul
Brain Korea 21 Project for Medical Science, Yonsei University, Seoul, Republic of Korea.
Eur J Pharmacol. 2008 May 31;586(1-3):226-33. doi: 10.1016/j.ejphar.2008.02.072. Epub 2008 Feb 29.
Vascular smooth muscle cell (VSMC) proliferation and migration in response to platelet-derived growth factor (PDGF) play an important role in the development of atherosclerosis and restenosis. Recent evidence indicates that PDGF increases intracellular levels of reactive oxygen species in VSMCs and that both PDGF-induced VSMC proliferation and migration are reactive oxygen species-dependent. Danshen is a representative oriental medicine used for the treatment of vascular disease. Previously, we reported that magnesium lithospermate B, an active component of Danshen, is a potent antioxidant. Thus we investigated the therapeutic potential of magnesium lithospermate B in neointimal formation after carotid artery injury in rats along with its effects on the PDGF signaling pathway for stimulating VSMC proliferation and migration in vitro. PDGF is dimeric glycoprotein composed of two A or two B chains. In this study, we used PDGF-BB, which is one of the isoforms of PDGF (i.e., PDGF-AA, PDGF-BB, and PDGF-AB). Our results demonstrated that magnesium lithospermate B directly scavenged reactive oxygen species in a xanthine/xanthine oxidase system and reduced PDGF-BB-induced intracellular reactive oxygen species generation in VSMCs. In a rat carotid artery balloon injury model, magnesium lithospermate B treatment (10 mg/kg/day, i.p) showed a significant effect on the prevention of neointimal formation compared with vehicle treatment. In cultured VSMCs, magnesium lithospermate B significantly attenuated PDGF-BB-induced cell proliferation and migration as measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2-tetrazolium bromide (MTT) assay and transwell migration assays, respectively. Further, magnesium lithospermate B inhibited PDGF-BB-induced phosphorylation of phospatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways by scavenging reactive oxygen species. Together, these data indicated that magnesium lithospermate B, a potent reactive oxygen species scavenger, prevented both injury-induced neointimal formation in vivo and PDGF-BB-induced VSMC proliferation and migration in vitro, suggesting that magnesium lithospermate B may be a promising agent to prevent atherosclerosis and restenosis following angioplasty.
血管平滑肌细胞（VSMC）对血小板衍生生长因子（PDGF）作出反应而发生的增殖和迁移，在动脉粥样硬化和再狭窄的发展过程中起着重要作用。最近的证据表明，PDGF会增加VSMC内活性氧的水平，而且PDGF诱导的VSMC增殖和迁移均依赖于活性氧。丹参是一种用于治疗血管疾病的典型传统中药。此前，我们报道过丹参的活性成分丹酚酸B镁是一种强效抗氧化剂。因此，我们研究了丹酚酸B镁对大鼠颈动脉损伤后新生内膜形成的治疗潜力，以及它对体外刺激VSMC增殖和迁移的PDGF信号通路的影响。PDGF是一种由两条A链或两条B链组成的二聚体糖蛋白。在本研究中，我们使用了PDGF-BB，它是PDGF的异构体之一（即PDGF-AA、PDGF-BB和PDGF-AB）。我们的结果表明，丹酚酸B镁在黄嘌呤/黄嘌呤氧化酶系统中直接清除活性氧，并减少了PDGF-BB诱导的VSMC内活性氧的生成。在大鼠颈动脉球囊损伤模型中，与溶剂处理相比，丹酚酸B镁处理（10毫克/千克/天，腹腔注射）对预防新生内膜形成具有显著效果。在培养的VSMC中，丹酚酸B镁分别通过3-[4,5-二甲基-2-噻唑基]-2,5-二苯基-2-溴化四氮唑（MTT）法和Transwell迁移试验测定，显著减弱了PDGF-BB诱导的细胞增殖和迁移。此外，丹酚酸B镁通过清除活性氧，抑制了PDGF-BB诱导的磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）和丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）通路的磷酸化。总之，这些数据表明，强效活性氧清除剂丹酚酸B镁既可以预防体内损伤诱导的新生内膜形成，也可以预防体外PDGF-BB诱导的VSMC增殖和迁移，这表明丹酚酸B镁可能是一种预防血管成形术后动脉粥样硬化和再狭窄的有前景的药物。
