Wang Xinwen, Liang Yan, Shi Jian, Zhu Hao-Jie, Bleske Barry E
Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, Ann Arbor, Michigan, USA.
The Key Laboratory of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing, China.
Planta Med. 2018 Oct;84(15):1094-1100. doi: 10.1055/a-0601-7083. Epub 2018 Apr 16.
Increased expression of microRNA 155 (miR-155) results in a decrease in endothelial nitric oxide synthase (eNOS) expression and impaired endothelial function. Factors that have been shown to increase expression of miR-155 may be mitigated by WS 1442, an extract of hawthorn leaves and flowers ( special extract) that contains a range of pharmacologically active substances including oligomeric proanthocyanidins and flavonoids. The purpose of this study is to determine the effect of WS 1442 on the expression of miR-155 and eNOS in the presence of tumor necrosis factor (TNF-). Human umbilical vein endothelial cells (HUVECs) were studied after the exposure to TNF-, with or without simvastatin (positive control) and WS 1442. The expression levels of eNOS, phosphorylated eNOS, and miR-155 in the different HUVEC treatment groups were determined by western blot and quantitative real-time polymerase chain reaction, respectively. To evaluate the effect of WS 1442 on the eNOS activity, the medium and intracellular nitrate/nitrite (NO) concentrations were also analyzed using a colorimetric Griess assay kit. The results demonstrated that TNF- upregulated miR-155 expression and decreased eNOS expression and NO concentrations. WS 1442 also increased miR-155 expression and decreased eNOS expression but, unlike TNF-, increased phosphorylated eNOS expression and NO concentrations. Surprisingly, WS 1442 increased miR-155 expression; however, WS 1442 mitigated the overall negative effect of miR-155 on decreasing eNOS expression by increasing expression of phosphorylated eNOS and resulting in an increase in NO concentrations. In the setting where miR-155 may be expressed, WS 1442 may offer vascular protection by increasing the expression of phosphorylated eNOS.
微小RNA 155（miR - 155）表达增加会导致内皮型一氧化氮合酶（eNOS）表达减少以及内皮功能受损。已证明可增加miR - 155表达的因素，可能会被WS 1442减轻，WS 1442是山楂叶和花的提取物（特殊提取物），含有一系列药理活性物质，包括低聚原花青素和黄酮类化合物。本研究的目的是确定在存在肿瘤坏死因子（TNF -）的情况下，WS 1442对miR - 155和eNOS表达的影响。在暴露于TNF - 后，对人脐静脉内皮细胞（HUVECs）进行研究，同时添加或不添加辛伐他汀（阳性对照）和WS 1442。分别通过蛋白质印迹法和定量实时聚合酶链反应测定不同HUVEC处理组中eNOS、磷酸化eNOS和miR - 155的表达水平。为了评估WS 1442对eNOS活性的影响，还使用比色Griess检测试剂盒分析培养基和细胞内硝酸盐/亚硝酸盐（NO）浓度。结果表明，TNF - 上调了miR - 155表达，降低了eNOS表达和NO浓度。WS 1442也增加了miR - 155表达并降低了eNOS表达，但与TNF - 不同的是，它增加了磷酸化eNOS表达和NO浓度。令人惊讶的是，WS 1442增加了miR - 155表达；然而，WS 1442通过增加磷酸化eNOS表达并导致NO浓度增加，减轻了miR - 155对降低eNOS表达的总体负面影响。在可能表达miR - 155的情况下，WS 1442可能通过增加磷酸化eNOS的表达来提供血管保护。
