Direct Effects of Empagliflozin on Extracellular Matrix Remodelling in Human Cardiac Myofibroblasts: Novel Translational Clues to Explain EMPA-REG OUTCOME Results
Direct Effects of Empagliflozin on Extracellular Matrix Remodelling in Human Cardiac Myofibroblasts: Novel Translational Clues to Explain EMPA-REG OUTCOME Results
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DOI：
10.1016/j.cjca.2019.08.033
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发表时间：
2020-04-01
影响因子：
6.2
通讯作者：
Fedak, Paul W. M.
中科院分区：
文献类型：
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作者：
Kang, Sean;Verma, Subodh;Fedak, Paul W. M.
背景资料：恩格列净是一种SGLT 2抑制剂，已显示心血管死亡率和心力衰竭住院率显著降低（EMPA-REG OUTCOME）。然而，恩格列净的心力衰竭保护作用的潜在机制在很大程度上仍然未知。心脏成纤维细胞在结构性心脏重塑和心力衰竭的进展中发挥不可或缺的作用，部分是通过调节细胞外基质（ECM）的稳态。本研究的目的是确定恩格列净是否对人心肌成肌纤维细胞介导的ECM remodeling.Methods：心脏成纤维细胞通过体外培养从心内直视手术获得的人心房组织中分离。胶原凝胶收缩试验用于评估肌成纤维细胞活性。使用共聚焦显微镜检查细胞形态和细胞介导的ECM重塑。使用逆转录定量聚合酶链反应评估促纤维化标志物的基因表达。结果：恩格列净暴露72小时后，通过胶原凝胶收缩显著减弱转化生长因子β 1诱导的成纤维细胞活化，浓度升高（0.5 μ mol/L，1 μ mol/L和5 μ mol/L）导致更大的衰减。形态学评估显示，暴露于恩格列净的肌成纤维细胞尺寸较小，延伸较短且数量较少，表明其表型更为静止。此外，通过胶原纤维排列指数测量，恩格列净显著减弱了细胞介导的ECM重塑。基因表达谱分析显示恩格列净显著抑制了关键的促纤维化标志物，包括COL 1A 1、ACTA 2、CTGF、FN 1和MMP-2。结论：我们提供了新的数据，表明恩格列净通过减弱肌成纤维细胞活性和细胞介导的胶原重塑对人心脏肌成纤维细胞表型和功能产生直接影响。这些数据为EMPA-REG OUTCOME研究中发现的恩格列净的深远影响提供了重要见解。
Background: Empagliflozin, an SGLT2 inhibitor, has shown remarkable reductions in cardiovascular mortality and heart failure admissions (EMPA-REG OUTCOME). However, the mechanism underlying the heart failure protective effects of empagliflozin remains largely unknown. Cardiac fibroblasts play an integral role in the progression of structural cardiac remodelling and heart failure, in part, by regulating extracellular matrix (ECM) homeostasis. The objective of this study was to determine if empagliflozin has a direct effect on human cardiac myofibroblast-mediated ECM remodelling.Methods: Cardiac fibroblasts were isolated via explant culture from human atrial tissue obtained at open heart surgery. Collagen gel contraction assay was used to assess myofibroblast activity. Cell morphology and cell-mediated ECM remodelling was examined with the use of confocal microscopy. Gene expression of profibrotic markers was assessed with the use of reverse-transcription quantitative polymerase chain reaction.Results: Empagliflozin significantly attenuated transforming growth factor beta 1-induced fibroblast activation via collagen gel contraction after 72-hour exposure, with escalating concentrations (0.5 mu mol/L, 1 mu mol/L, and 5 mu mol/L) resulting in greater attenuation. Morphologic assessment showed that myofibroblasts exposed to empagliflozin were smaller in size with shorter and fewer number of extensions, indicative of a more quiescent phenotype. Moreover, empagliflozin significantly attenuated cell-mediated ECM remodelling as measured by collagen fibre alignment index. Gene expression profiling revealed significant suppression of critical profibrotic markers by empagliflozin, including COL1A1, ACTA2, CTGF, FN1, and MMP-2.Conclusions: We provide novel data showing a direct effect of empagliflozin on human cardiac myofibroblast phenotype and function by attenuation of myofibroblast activity and cell-mediated collagen remodelling. These data provide critical insights into the profound effects of empagliflozin as noted in the EMPA-REG OUTCOME study.