丹参酮I通过靶向TGFBR1，调节TGF-β信号通路减轻缺血后心肌损伤

IF 6.7 1区医学 Q1 CHEMISTRY, MEDICINAL

Phytomedicine Pub Date : 2025-06-16 DOI:10.1016/j.phymed.2025.156994

Liyuan Ke , Ziyao Zheng , Shumin Ye , Chenhui Zhong , Qingyun Lin , Yan Hu , Peiying Shi , Lei Wen , Hong Yao

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引用次数: 0

摘要

丹参酮I （tanshinone I, Tan I）是传统心血管药物丹参（S. miltiorrhiza Bunge）的重要活性成分。Tan I对心肌细胞的保护作用已有报道，但其抗心肌缺血作用及其机制尚不清楚。目的系统评价Tan I减轻心肌缺血（MI）损伤的作用，并阐明Tan I改善小鼠心肌纤维化和心室功能的潜在分子机制。方法通过构建体内和体外心肌梗死模型，验证谭一抗心肌梗死的作用，通过靶点捕捞、分子对接和网络药理学研究，进一步预测谭一抗心肌梗死的作用机制和潜在靶点蛋白。随后，采用基于TMT的定量蛋白质组学、转化生长因子β受体1 （TGFBR1）过表达慢病毒载体、分子动力学（MD）模拟、生物层干涉法（BLI）、细胞热移法（CETSA）、TGFBR1激酶活性和药物亲和力反应靶稳定性（DARTS）实验验证了Tan I的抗mi效应机制和靶点。改善细胞内环境，抑制细胞内活性氧的释放。在心肌梗死小鼠模型中具有恢复异常心电图、减少心肌梗死面积、抑制心肌纤维化、降低血清关键心脏损伤生物标志物水平的作用。在机制上，Tan I显著抑制了心肌梗死小鼠TGFBR1和Smad2的磷酸化修饰水平，以及I/III胶原蛋白、α-平滑肌肌动蛋白、Bcl-2和Bax蛋白的异常表达。这些发现在过表达TGFBR1或被TGF-β1激活的NIH-3T3细胞中得到进一步验证。MD模拟、CETSA和dart显示TGFBR1与Tan I的结合相对稳定。此外，BLI表明Tan I结合TGFBR1的平衡解离常数为1.5 × 10−6 m。基于激酶活性测定，Tan I抑制TGFBR1的半最大抑制浓度为739.6 nM。结论本研究首次揭示了Tan I通过靶向TGFBR1调节TGF-β信号通路，从而减轻心肌梗死损伤和纤维化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Tanshinone I alleviates post-ischemic myocardial injury by targeting TGFBR1 and modulating the TGF-β signaling pathway

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Tanshinone I alleviates post-ischemic myocardial injury by targeting TGFBR1 and modulating the TGF-β signaling pathway

Background

Tanshinone I (Tan I) is an essential active ingredient of the traditional cardiovascular medicine Salvia miltiorrhiza Bunge (S. miltiorrhiza). Although the protection of Tan I on cardiomyocyte has been reported, its anti-myocardial ischemia effects and mechanisms remain unknown.

Purpose

Systematic evaluation of the role of Tan I in reducing myocardial ischemia (MI) injury and elucidation of the underlying molecular mechanisms by which Tan I improves myocardial fibrosis and ventricular function in mouse MI models.

Methods

In vivo and in vitro MI models were constructed to substantiate the anti-MI effects of Tan I. Through target fishing, molecular docking, and network pharmacology investigation, the effect mechanisms and potential target proteins of Tan I against MI were predicted further. Tandem mass tags (TMT)-based quantitative proteomics, transforming growth factor beta receptor I (TGFBR1)-overexpressing lentiviral vectors, molecular dynamics (MD) simulations, biolayer interferometry (BLI), cellular thermal shift assay (CETSA), TGFBR1 kinase activity, and drug affinity responsive target stability (DARTS) assay were subsequently used to validate the anti-MI-effect mechanisms and targets of Tan I.

Results

Tan I can markedly increase the survival of oxidative stress cell models, improve intracellular environment, and inhibit the release of intracellular reactive oxygen species. Moreover, it can restore abnormal electrocardiograms, decrease myocardial infarction area, inhibit cardiac fibrosis, and reduce serum levels of key cardiac injury biomarkers in the MI mouse model. Mechanistically, Tan I considerably inhibited the phosphorylation modification levels of TGFBR1 and Smad2 and the aberrant expressions of Collagen I/III, α-smooth muscle actin, Bcl-2, and Bax proteins in MI mice. These findings were further verified in NIH-3T3 cells overexpressing TGFBR1 or activated by TGF-β1. MD simulations, CETSA, and DARTS showed that TGFBR1 binding to Tan I was relatively stable. In addition, BLI indicated that the equilibrium dissociation constant of Tan I binding TGFBR1 was 1.5 × 10⁻⁶ M. Based on the kinase activity assay, Tan I restrained TGFBR1 with a half-maximal inhibitory concentration of 739.6 nM.

Conclusion

This work reveals for the first time that Tan I can reduce MI injury and fibrosis by modulating the TGF-β signaling pathway via targeting of TGFBR1.

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来源期刊

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.