熊果苷通过激活 AMPK/CREB/PGC⁃1α 通路减轻心肌梗死后心脏重构

Arbutin alleviates cardiac remodeling after myocardial infarction by activating the AMPK/CREB/PGC⁃1α pathway

摘要

目的：探讨熊果苷（Arbutin）对心肌梗死（MI）后心脏重构（MR）的治疗作用及其分子机制。方法：采用左前降支结扎法建立小鼠 MI 模型，40 只 8 周龄雄性 C57BL/6 小鼠随机分为 Sham+Vehicle 组、Sham+Arbutin 组、MI+Vehicle 组及 MI+Arbutin 组（n=10）。术后 24 h 开始腹腔注射熊果苷（10 mg·kg^-1·d^-1）或等体积生理盐水，连续 4 周。超声心动图评估心脏结构与功能，测定血清心肌损伤标志物。体外实验采用新生大鼠心肌细胞（NRCMs）构建缺氧模型，随机计数法分为 Norm+Vehicle 组、Norm+Arbutin 组、Hypo+Vehicle 组及 Hypo+Arbutin 组。采用 Western Blot、RT⁃qPCR 和组织学方法分析分子机制，通过药物抑制实验和基因敲低实验验证 AMPK 通路的作用。结果：与 MI 组相比，熊果苷治疗能够改善 MI 后左室射血分数和缩短分数，缩小梗死面积，降低血清 cTnT、CK⁃MB 和 LDH 水平（P<0.000 1）。组织学检查显示熊果苷可减轻心肌细胞肥大、间质纤维化、氧化应激和细胞凋亡（P<0.000 1）。机制研究表明，熊果苷可增强 AMPKα 磷酸化，激活 CREB 并上调 PGC⁃1α 表达（P<0.000 1）。体外实验证实熊果苷能够促进缺氧心肌细胞存活，抑制肥厚标志物 ANP 和 BNP 表达（P<0.000 1）。应用 AMPK 抑制剂 Compound C 或 siRNA 敲低 AMPKα 后，熊果苷的保护作用被抵消。结论：熊果苷能够通过激活 AMPK/CREB/PGC⁃1α 信号通路减轻心肌梗死后心脏重构，是防治 MI 后心力衰竭的潜在治疗药物。

Abstract

Objective: To explore the therapeutic effect of arbutin on myocardial remodeling (MR) after myocardial infarction (MI) and its molecular mechanism. Methods: The mouse MI model was established by ligation of the left anterior descending branch. Forty 8⁃week⁃old male C57BL/6 mice were randomly divided into the Sham+Vehicle group, the Sham+Arbutin group, the MI+Vehicle group, and the MI+Arbutin group (n=10). Arbutin (10 mg·kg^-1·d^-1) or an equal volume of normal saline was intraperitoneally injected 24 hours after the operation for 4 consecutive weeks. Echocardiography was used to evaluate the structure and function of the heart and determine serum myocardial injury markers. In vitro experiments were conducted using neonatal rat cardiomyocytes (NRCMs) to construct a hypoxia model, which was randomly divided into the Norm+Vehicle group, Norm+Arbutin group, Hypo+Vehicle group, and Hypo+Arbutin group by the random counting method. The molecular mechanism was analyzed by Western Blot, RT⁃qPCR and histological methods, and the role of the AMPK pathway was verified by drug inhibition experiments and gene knockdown experiments. Results: Compared with the MI group, Arbutin treatment could improve the left ventricular ejection fraction and shortening fraction after myocardial infarction, reduce the infarction area, and lower the levels of serum cTnT, CK⁃MB, and LDH (P<0.000 1). Histological examination showed that Arbutin could alleviate myocardial cell hypertrophy, interstitial fibrosis, oxidative stress and apoptosis (P<0.000 1). Mechanism studies showed that Arbutin enhanced the phosphorylation of AMPKα, activated CREB and up⁃regulated the expression of PGC⁃1α (P<0.000 1).In vitro experiments confirmed that Arbutin could improve the survival of hypoxic cardiomyocytes and inhibit the expression of hypertrophic markers ANP and BNP (P<0.000 1). After knocking down AMPKα with AMPK inhibitors Compound C or siRNA, the protective effect of arbutin was eliminated. Conclusion: Arbutin can alleviate cardiac remodeling after myocardial infarction by activating the AMPK/CREB/PGC⁃1α signaling pathway and is a potential therapeutic drug for the prevention and treatment of heart failure after MI.

关键词

熊果苷 / 心肌梗死 / 心脏重构 / 腺苷酸活化蛋白激酶 / 过氧化物酶体增殖物激活受体γ 辅激活因子⁃1α

Key words

Arbutin / Myocardial Infarction / Myocardial Remodeling / AMPK / PGC⁃1α

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基金资助

湖北省自然科学基金项目资助项目(2023AFC027)