背景:内皮功能障碍(ED),以一氧化氮(NO)生物利用度明显降低为特征,血管收缩,向促炎和血栓形成状态转变,是高血压的重要原因,动脉粥样硬化,和其他心血管疾病。腺苷5'-单磷酸(AMP)激活的蛋白激酶(AMPK)广泛参与心血管发育。PrzewaquinoneA(PA),从丹参中提取的亲脂性二萜醌,抑制血管收缩。
目的:在此,目的是探讨PA在体内和体外对ED的保护作用,以及潜在的机制。
方法:采用血管紧张素II(AngII)诱导的人脐静脉内皮细胞(HUVEC)模型进行体外观察。AMPK水平,内皮型一氧化氮合酶(eNOS),血管细胞粘附分子-1(VCAM-1),一氧化氮(NO),免疫印迹法和ELISA法检测内皮素-1(ET-1)。采用渗透泵连续输注AngII(1000ng/kg/min)4周建立小鼠高血压模型。PA和/或缬沙坦给药后,测定NO和ET-1水平。通过免疫组织化学评估胸主动脉中AMPK信号传导相关蛋白的水平。收缩压(SBP),舒张压(DBP),使用尾套法测量平均动脉压(MAP)。分离的主动脉血管张力测量用于评估小鼠的血管舒张功能。分子对接,分子动力学,和表面等离子体共振成像(SPRi)用于确认AMPK和PA的相互作用。
结果:PA以剂量依赖性方式抑制AngII诱导的血管收缩和血管粘附以及激活的AMPK信号传导。此外,PA显著抑制血压,AngII刺激后小鼠血管舒张激活,并促进AMPK信号的激活。此外,分子模拟和SPRi揭示了PA直接靶向AMPK。AMPK抑制部分消除了PA对内皮功能障碍的保护作用。
结论:PA激活AMPK并改善高血压期间的内皮功能障碍。
BACKGROUND: Endothelial dysfunction (ED), characterized by markedly reduced nitric oxide (NO) bioavailability, vasoconstriction, and a shift toward a proinflammatory and prothrombotic state, is an important contributor to hypertension, atherosclerosis, and other cardiovascular diseases. Adenosine 5\'-monophosphate (AMP)-activated protein kinase (AMPK) is widely involved in cardiovascular development. Przewaquinone A (PA), a lipophilic diterpene quinone extracted from Salvia przewalskii Maxim, inhibits vascular contraction.
OBJECTIVE: Herein, the goal was to explore the protective effect of PA on ED in vivo and in vitro, as well as the underlying mechanisms.
METHODS: A human umbilical vein endothelial cell (HUVEC) model of ED induced by angiotensin II (AngII) was used for in vitro observations. Levels of AMPK, endothelial nitric oxide synthase (eNOS), vascular cell adhesion molecule-1 (VCAM-1), nitric oxide (NO), and endothelin-1 (ET-1) were detected by western blotting and ELISA. A mouse model of hypertension was established by continuous infusion of AngII (1000 ng/kg/min) for 4 weeks using osmotic pumps. Following PA and/or valsartan administration, NO and ET-1 levels were measured. The levels of AMPK signaling-related proteins in the thoracic aorta were evaluated by immunohistochemistry. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) were measured using the tail cuff method. Isolated aortic vascular tone measurements were used to evaluate the vasodilatory function in mice. Molecular docking, molecular dynamics, and surface plasmon resonance imaging (SPRi) were used to confirm AMPK and PA interactions.
RESULTS: PA inhibited AngII-induced vasoconstriction and vascular adhesion as well as activated AMPK signaling in a dose-dependent manner. Moreover, PA markedly suppressed blood pressure, activated vasodilation in mice following AngII stimulation, and promoted the activation of AMPK signaling. Furthermore, molecular simulations and SPRi revealed that PA directly targeted AMPK. AMPK inhibition partly abolished the protective effects of PA against endothelial dysfunction.
CONCLUSIONS: PA activates AMPK and ameliorates endothelial dysfunction during hypertension.