槲皮素通过促进 miR-135b 的表达抑制 TGF-β/Smads 通路改善心房颤动。
Quercetin improves atrial fibrillation through inhibiting TGF-β/Smads pathway via promoting MiR-135b expression.
机构信息
Department of Cardiology, The Second Affiliated Hospital of Xi'an JiaoTong University, China.
Department of Cardiology, The Second Affiliated Hospital of Xi'an JiaoTong University, China.
出版信息
Phytomedicine. 2021 Dec;93:153774. doi: 10.1016/j.phymed.2021.153774. Epub 2021 Sep 26.
PURPOSE
To investigate the role and mechanism of quercetin in isoprenaline (ISO)-induced atrial fibrillation (AF).
STUDY DESIGN
Rat cardiac fibroblasts (RCFs) models and RCFs were used to explore the effect and underlying mechanism of quercetin in isoprenaline (ISO)-induced atrial fibrillation (AF) in vivo and in vitro by a series of experiments.
METHODS
Differentially expressed microRNAs were screened from human AF tissues using the GEO2R and RT-qPCR. The expressions of TGF-β/Smads pathway molecules (TGFβ1, TGFBR1, Tgfbr1, Tgfbr2, Smad2, Smad3, Smad4) in AF tissues were detected by RT-qPCR and Western blot. The relationships between miR-135b and genes (Tgfbr1, Tgfbr2, Smad2) were analyzed by Pearson correlation, TargetScan and dual-luciferase activity assay. RCFs induced by ISO were treated with quercetin (20 or 50 μM), miR-135b mimic and inhibitor, siTgfbr1 and their corresponding controls, then the cell viability was determined by MTT and the expressions of cyclin D1, α-SMA, collagen-related molecules, TGF-β/Smads pathway molecules, and miR-135b were measured by RT-qPCR and Western blot. ISO-induced rats were treated with quercetin (25 mg/kg/day) via gavage, miR-135b antagomir, agomir and their corresponding controls. The treated rats were used for the detection of miR-135b expression by RT-qPCR, histopathological observation by HE and Masson staining, and the detection of Col1A1 and fibronectin contents by immunohistochemical technique.
RESULTS
The expression of miR-135b was downregulated, and those of TGFBR1, TGFBR2, target genes of miR-135b were upregulated in human AF tissues and negatively regulated by miR-135b in RCFs. Through inhibiting TGF-β/Smads pathway via promoting miR-135b expression, quercetin treatment inhibited proliferation, myofibroblast differentiation and collagen deposition in ISO-treated RCFs, as evidenced by reduced expressions of cyclin D1, α-SMA, collagen-related genes and proteins, and alleviated fibrosis and collagen deposition of atrial tissues in ISO-treated rats.
CONCLUSION
Quercetin may alleviate AF by inhibiting fibrosis of atrial tissues through inhibiting TGF-β/Smads pathway via promoting miR-135b expression.
目的
研究槲皮素在异丙肾上腺素（ISO）诱导的心房颤动（AF）中的作用和机制。
研究设计
使用大鼠心肌成纤维细胞（RCFs）模型和 RCFs，通过一系列实验探讨槲皮素在体内和体外 ISO 诱导的 AF 中的作用及潜在机制。
方法
利用 GEO2R 和 RT-qPCR 从人 AF 组织中筛选差异表达的 microRNAs。通过 RT-qPCR 和 Western blot 检测 AF 组织中 TGF-β/Smads 通路分子（TGFβ1、TGFBR1、Tgfbr1、Tgfbr2、Smad2、Smad3、Smad4）的表达。通过 Pearson 相关性分析、TargetScan 和双荧光素酶活性测定分析 miR-135b 与基因（Tgfbr1、Tgfbr2、Smad2）的关系。用 ISO 诱导 RCFs，然后用槲皮素（20 或 50 μM）、miR-135b 模拟物和抑制剂、siTgfbr1 及其相应的对照物处理，然后用 MTT 法测定细胞活力，用 RT-qPCR 和 Western blot 法测定细胞周期蛋白 D1、α-SMA、胶原相关分子、TGF-β/Smads 通路分子和 miR-135b 的表达。通过灌胃给予 ISO 诱导的大鼠槲皮素（25 mg/kg/天）、miR-135b 拮抗剂、激动剂及其相应的对照物。用 RT-qPCR 检测大鼠的 miR-135b 表达，用 HE 和 Masson 染色进行组织病理学观察，用免疫组织化学技术检测 Col1A1 和纤维连接蛋白含量。
结果
miR-135b 在人 AF 组织中表达下调，而 TGFBR1、TGFBR2、miR-135b 的靶基因在 RCFs 中上调，并受 miR-135b 的负调控。通过促进 miR-135b 的表达抑制 TGF-β/Smads 通路，槲皮素治疗抑制了 ISO 处理的 RCFs 中的增殖、成纤维细胞分化和胶原沉积，表现为细胞周期蛋白 D1、α-SMA、胶原相关基因和蛋白表达减少，并减轻了 ISO 处理大鼠心房组织的纤维化和胶原沉积。
结论
槲皮素可能通过促进 miR-135b 的表达抑制 TGF-β/Smads 通路，从而抑制心房组织纤维化，减轻 AF。
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