Zhang Liyun, Chen Juan, Yan Lianhua, He Qin, Xie Han, Chen Manhua
Department of Cardiology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Front Pharmacol. 2021 Jun 10;12:646240. doi: 10.3389/fphar.2021.646240. eCollection 2021.
Accumulating evidence suggested that resveratrol (RES) could protect against adverse cardiac remodeling induced by several cardiovascular diseases. However, the role of RES in the setting of heart failure with preserved ejection fraction (HFpEF) and the underlying mechanisms of its action remain understood. This study was to determine whether RES could ameliorate HFpEF-induced cardiac remodeling and its mechanisms. , C57BL/6 mice served as either the sham or the HFpEF model. The HFpEF mice model was induced by uninephrectomy surgery and d-aldosterone infusion. RES (10 mg/kg/day, ig) or saline was administered to the mice for four weeks. , transforming growth factor β1 (TGF-β1) was used to stimulate neonatal rat cardiac fibroblasts (CFs) and Ex-527 was used to inhibit sirtuin 1 (Sirt1) in CFs. Echocardiography, hemodynamics, western blotting, quantitative real-time PCR, histological analysis, immunofluorescence, and ELISA kits were used to evaluate cardiac remodeling induced by HFpEF. Sirt1 and Smad3 expressions were measured to explore the underlying mechanisms of RES. HFpEF mice developed left ventricular hypertrophy, preserved ejection fraction, diastolic dysfunction, and pulmonary congestion. Moreover, HFpEF mice showed increased infiltration of neutrophils and macrophages into the heart, including increased interleukin (IL)-1β, IL-6, and TNF-α. We also observed elevated M1 macrophages and decreased M2 macrophages, which were exhibited by increased mRNA expression of M1 markers (iNOS, CD86, and CD80) and decreased mRNA expression of M2 markers (Arg1, CD163, and CD206) in HFpEF hearts. Moreover, HFpEF hearts showed increased levels of intracellular reactive oxygen species (ROS). Importantly, HFpEF mice depicted increased and and TGF-β mRNA expressions and decreased protein expression of phosphorylated endothelial nitric-oxide synthase (p-eNOS). Results of western blot revealed that the activated TGF-β/Smad3 signaling pathway mediated HFpEF-induced cardiac remodeling. As expected, this HFpEF-induced cardiac remodeling was reversed when treated with RES. RES significantly decreased Smad3 acetylation and inhibited Smad3 transcriptional activity induced by HFpEF activating Sirt1. Inhibited Sirt1 with Ex-527 increased Smad3 acetylation, enhanced Smad3 transcriptional activity, and offset the protective effect of RES on TGF-β-induced cardiac fibroblast-myofibroblast transformation in CFs. Our results suggested that RES exerts a protective action against HFpEF-induced adverse cardiac remodeling by decreasing Smad3 acetylation and transcriptional activity activating Sirt1. RES is expected to be a novel therapy option for HFpEF patients.
越来越多的证据表明，白藜芦醇（RES）可以预防由多种心血管疾病引起的不良心脏重塑。然而，RES在射血分数保留的心力衰竭（HFpEF）中的作用及其作用的潜在机制仍不清楚。本研究旨在确定RES是否可以改善HFpEF引起的心脏重塑及其机制。为此，将C57BL/6小鼠作为假手术组或HFpEF模型组。通过单侧肾切除手术和输注d-醛固酮诱导建立HFpEF小鼠模型。给小鼠灌胃RES（10mg/kg/天）或生理盐水，持续四周。此外，用转化生长因子β1（TGF-β1）刺激新生大鼠心脏成纤维细胞（CFs），并用Ex-527抑制CFs中的沉默调节蛋白1（Sirt1）。采用超声心动图、血流动力学、蛋白质免疫印迹、实时定量PCR、组织学分析、免疫荧光和ELISA试剂盒评估HFpEF诱导的心脏重塑。检测Sirt1和Smad3的表达以探究RES的潜在作用机制。HFpEF小鼠出现左心室肥厚、射血分数保留、舒张功能障碍和肺充血。此外，HFpEF小鼠心脏中中性粒细胞和巨噬细胞浸润增加，包括白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α增加。我们还观察到M1巨噬细胞增加而M2巨噬细胞减少，这表现为HFpEF心脏中M1标志物（诱导型一氧化氮合酶、CD86和CD80）的mRNA表达增加以及M2标志物（精氨酸酶1、CD163和CD206）的mRNA表达减少。此外，HFpEF心脏中细胞内活性氧（ROS）水平升高。重要的是，HFpEF小鼠的TGF-β1和TGF-β mRNA表达增加，而磷酸化内皮型一氧化氮合酶（p-eNOS）的蛋白表达减少。蛋白质免疫印迹结果显示，激活的TGF-β/Smad3信号通路介导了HFpEF诱导的心脏重塑。正如预期的那样，用RES治疗可逆转这种HFpEF诱导的心脏重塑。RES显著降低Smad3乙酰化水平，并通过激活Sirt1抑制HFpEF诱导的Smad3转录活性。用Ex-527抑制Sirt1会增加Smad3乙酰化水平，增强Smad3转录活性，并抵消RES对TGF-β诱导的CFs心脏成纤维细胞-肌成纤维细胞转化的保护作用。我们的研究结果表明，RES通过激活Sirt1降低Smad3乙酰化水平和转录活性，从而对HFpEF诱导的不良心脏重塑发挥保护作用。RES有望成为HFpEF患者的一种新型治疗选择。
