Liou Shu-Fen, Nguyen Thi Tuyet Ngan, Hsu Jong-Hau, Sulistyowati Erna, Huang Shang-En, Wu Bin-Nan, Lin Ming-Chung, Yeh Jwu-Lai
Department of Pharmacy, Chia-Nan University of Pharmacy and Science, Tainan 717, Taiwan.
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
Antioxidants (Basel). 2020 Oct 6;9(10):956. doi: 10.3390/antiox9100956.
Vascular calcification (VC) is highly prevalent in patients with atherosclerosis, chronic kidney disease, diabetes mellitus, and hypertension. In blood vessels, VC is associated with major adverse cardiovascular events. Xanthohumol (XN), a main prenylated chalcone found in hops, has antioxidant effects to inhibit VC. This study aimed to investigate whether XN attenuates VC through in vivo study. A rat VC model was established by four weeks oral administration of vitamin D plus nicotine in Sprague Dawley (SD) rats. In brief, 30 male SD rats were randomly divided into three groups: control, 25 mg/kg nicotine in 5 mL corn oil and 3 × 10 IU/kg vitamin D administration (VDN), and combination of VDN with 20 mg/L in 0.1% ethanol of XN (treatment group). Physiological variables such as body and heart weight and drinking consumption were weekly observed, and treatment with XN caused no differences among the groups. In comparison with the control group, calcium content and alkaline phosphatase (ALP) activity were increased in calcified arteries, and XN treatment reduced these levels. Dihydroethidium (DHE) and 2',7'-dichloroflurescin diacetate (DCFH-DA) staining to identify Superoxide and reactive oxygen species generation from aorta tissue showed increased production in VDN group compared with the control and treatment groups. Hematoxylin eosin (HE) and Alizarin Red S staining were determined to show medial vascular thickness and calcification of vessel wall. Administration of VDN resulted in VC, and XN treatment showed improvement in vascular structure. Moreover, overexpression of osteogenic transcription factors bone morphogenetic protein 2 (BMP-2) and runt-related transcription factor 2 (Runx2) were significantly suppressed by XN treatment in VC. Moreover, downregulation of vascular phenotypic markers alpha-smooth muscle actin (α-SMA) and smooth muscle 22 alpha (SM22α) were increased by XN treatment in VC. Furthermore, XN treatment in VC upregulated nuclear translocation of nuclear factor-E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expressions. Otherwise, Kelch-like ECH-associated protein 1 (Keap1) was alleviated by XN treatment in VC. In conclusion, our findings suggested that XN enhances antioxidant capacity to improve VC by regulating the Nrf2/Keap1/HO-1 pathway. Therefore, XN may have potential effects to decrease cardiovascular risk by reducing VC.
血管钙化（VC）在动脉粥样硬化、慢性肾病、糖尿病和高血压患者中极为常见。在血管中，VC与主要不良心血管事件相关。黄腐酚（XN）是啤酒花中发现的一种主要的异戊烯基查尔酮，具有抑制VC的抗氧化作用。本研究旨在通过体内研究探讨XN是否能减轻VC。通过对Sprague Dawley（SD）大鼠口服维生素D加尼古丁四周建立大鼠VC模型。简而言之，将30只雄性SD大鼠随机分为三组：对照组、5 mL玉米油中含25 mg/kg尼古丁和3×10 IU/kg维生素D给药组（VDN组）以及VDN与0.1%乙醇中20 mg/L XN的联合给药组（治疗组）。每周观察体重、心脏重量和饮水量等生理变量，XN治疗在各组之间未引起差异。与对照组相比，钙化动脉中的钙含量和碱性磷酸酶（ALP）活性增加，XN治疗降低了这些水平。用二氢乙锭（DHE）和2',7'-二氯荧光素二乙酸酯（DCFH-DA）染色来鉴定主动脉组织中产生的超氧化物和活性氧，结果显示与对照组和治疗组相比，VDN组的产生增加。苏木精伊红（HE）染色和茜素红S染色用于显示血管中层厚度和血管壁钙化。给予VDN导致VC，XN治疗显示血管结构有所改善。此外，XN治疗在VC中显著抑制了成骨转录因子骨形态发生蛋白2（BMP-2）和 runt相关转录因子2（Runx2）的过表达。此外，XN治疗在VC中使血管表型标志物α-平滑肌肌动蛋白（α-SMA）和平滑肌22α（SM22α）的下调增加。此外，XN治疗在VC中上调了核因子E2相关因子2（Nrf2）的核转位和血红素加氧酶-1（HO-1）的表达。另外，XN治疗降低了VC中kelch样ECH相关蛋白1（Keap1）的表达。总之，我们的研究结果表明，XN通过调节Nrf2/Keap1/HO-1途径增强抗氧化能力以改善VC。因此，XN可能具有通过减少VC来降低心血管风险的潜在作用。
