Izumi Yasukatsu, Shiota Masayuki, Kusakabe Hiromi, Hikita Yuko, Nakao Takafumi, Nakamura Yasuhiro, Muro Takashi, Miura Katsuyuki, Yoshiyama Minoru, Iwao Hiroshi
Department of Pharmacology, Osaka City University Medical School, Osaka 545-8585, Japan.
Hypertens Res. 2009 Aug;32(8):675-9. doi: 10.1038/hr.2009.77. Epub 2009 Jun 5.
Statins exert pleiotropic effects on the cardiovascular system, in part through an increase in nitric oxide (NO) bioavailability. In this study, we examined the role of pravastatin in ischemia-induced angiogenesis. Unilateral hindlimb ischemia was surgically induced in C57BL/6J mice. Phosphorylation of AMP-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC) and endothelial NO synthase (eNOS) was increased in ischemic tissues. Furthermore, mice treated with pravastatin showed higher increases in phosphorylation than did untreated mice. Laser Doppler analysis has shown that pravastatin treatment accelerates the development of collateral vessels and angiogenesis in response to hindlimb ischemia. Capillary density in the ischemic hindlimb was also increased by pravastatin treatment. An in vitro study on human umbilical vein endothelial cells (HUVECs) revealed that pravastatin increased the phosphorylation of AMPK. Pravastatin-induced phosphorylation of eNOS, one of the downstreams of AMPK, was inhibited by compound C, an AMPK antagonist. The increased migration and tube formation of HUVECs by pravastatin were significantly blocked by compound C treatment. The accelerated angiogenesis by pravastatin after hindlimb ischemia was significantly reduced after treatment with compound C. Thus, ischemia induced AMPK phosphorylation in vivo. Furthermore, pravastatin could also activate AMPK in vivo and in vitro. Such phosphorylation results in eNOS activation and angiogenesis, which provide a novel explanation for one of the pleiotropic effects of statins that is beneficial for angiogenesis.
他汀类药物对心血管系统具有多效性作用，部分是通过增加一氧化氮（NO）的生物利用度来实现的。在本研究中，我们研究了普伐他汀在缺血诱导的血管生成中的作用。通过手术诱导C57BL/6J小鼠单侧后肢缺血。缺血组织中AMP激活的蛋白激酶（AMPK）、乙酰辅酶A羧化酶（ACC）和内皮型一氧化氮合酶（eNOS）的磷酸化增加。此外，与未治疗的小鼠相比，接受普伐他汀治疗的小鼠磷酸化水平升高幅度更大。激光多普勒分析表明，普伐他汀治疗可加速侧支血管的发育以及对后肢缺血的血管生成反应。普伐他汀治疗还增加了缺血后肢的毛细血管密度。一项对人脐静脉内皮细胞（HUVECs）的体外研究表明，普伐他汀增加了AMPK的磷酸化。普伐他汀诱导的eNOS（AMPK的下游之一）磷酸化被AMPK拮抗剂化合物C抑制。普伐他汀导致的HUVECs迁移和管腔形成增加被化合物C处理显著阻断。后肢缺血后普伐他汀加速的血管生成在化合物C处理后显著降低。因此，缺血在体内诱导了AMPK磷酸化。此外，普伐他汀在体内和体外均可激活AMPK。这种磷酸化导致eNOS激活和血管生成，这为他汀类药物有利于血管生成的多效性作用之一提供了新的解释。
