Division of Cardiology, Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 704, Taiwan.
Department of Biological Science, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
Int J Mol Sci. 2022 Nov 27;23(23):14839. doi: 10.3390/ijms232314839.
Cilostazol is an antiplatelet agent with vasodilating effects that functions by increasing the intracellular concentration of cyclic adenosine monophosphate. We have previously shown that cilostazol has favorable effects on angiogenesis. However, there is no study to evaluate the effects of cilostazol on adiponectin. We investigated the effects of cilostazol on angiogenesis in diabetes in vitro and in vivo through adiponectin/adiponectin receptors (adipoRs) and the sirtuin 1 (SIRT1)/AMP-activated protein kinase (AMPK) signaling pathway. Human umbilical vein endothelial cells (HUVECs) and human aortic smooth muscle cells (HASMCs) were cocultured under high glucose (HG) conditions. Adiponectin concentrations in the supernatants were significantly increased when HASMCs were treated with cilostazol but not significantly changed when only HUVECs were treated with cilostazol. Cilostazol treatment enhanced the expression of SIRT1 and upregulated the phosphorylation of AMPK in HG-treated HUVECs. By sequential knockdown of adipoRs, SIRT1, and AMPK, our data demonstrated that cilostazol prevented apoptosis and stimulated proliferation, chemotactic motility, and capillary-like tube formation in HG-treated HUVECs through the adipoRs/SIRT1/AMPK signaling pathway. The phosphorylation of downstream signaling molecules, including acetyl-CoA carboxylase (ACC) and endothelial nitric oxide synthase (eNOS), was downregulated when HUVECs were treated with a SIRT1 inhibitor. In streptozotocin-induced diabetic mice, cilostazol treatment could improve blood flow recovery 21-28 days after inducing hindlimb ischemia as well as increase the circulating of CD34CD45 cells 14-21 days after operation; moreover, these effects were significantly attenuated by the knockdown of adipoR1 but not adipoR2. The expression of SIRT1 and phosphorylation of AMPK/ACC and Akt/eNOS in ischemic muscles were significantly attenuated by the gene knockdown of adipoRs. Cilostazol improves HG-induced endothelial dysfunction in vascular endothelial cells and enhances angiogenesis in diabetic mice by upregulating the expression of adiponectin/adipoRs and its SIRT1/AMPK downstream signaling pathway.
西洛他唑是一种具有血管扩张作用的抗血小板药物，通过增加环磷酸腺苷单磷酸的细胞内浓度而起作用。我们之前已经表明，西洛他唑对血管生成有良好的作用。然而，目前还没有研究评估西洛他唑对脂联素的影响。我们通过脂联素/脂联素受体（adipoRs）和沉默调节蛋白 1（SIRT1）/腺苷酸活化蛋白激酶（AMPK）信号通路，在体外和体内研究了西洛他唑对糖尿病血管生成的影响。将人脐静脉内皮细胞（HUVECs）和人主动脉平滑肌细胞（HASMCs）在高葡萄糖（HG）条件下共培养。当 HASMCs 用西洛他唑处理时，上清液中的脂联素浓度显著增加，而当仅用 HUVECs 处理时，脂联素浓度没有显著变化。西洛他唑处理增强了 HG 处理的 HUVECs 中 SIRT1 的表达，并上调了 AMPK 的磷酸化。通过顺序敲低 adipoRs、SIRT1 和 AMPK，我们的数据表明，西洛他唑通过 adipoRs/SIRT1/AMPK 信号通路防止了 HG 处理的 HUVECs 的凋亡，并刺激了增殖、趋化运动和毛细血管样管形成。当 HUVECs 用 SIRT1 抑制剂处理时，下游信号分子，包括乙酰辅酶 A 羧化酶（ACC）和内皮型一氧化氮合酶（eNOS）的磷酸化被下调。在链脲佐菌素诱导的糖尿病小鼠中，西洛他唑治疗可改善后肢缺血后 21-28 天的血流恢复，并增加术后 14-21 天的循环 CD34CD45 细胞；此外，这些作用被 adipoR1 的敲低显著减弱，但 adipoR2 没有减弱。缺血肌肉中 adipoRs 的基因敲低显著减弱了 SIRT1 和 AMPK/ACC 和 Akt/eNOS 的磷酸化。西洛他唑通过上调脂联素/脂联素受体及其 SIRT1/AMPK 下游信号通路，改善 HG 诱导的血管内皮细胞功能障碍，并增强糖尿病小鼠的血管生成。
