紫花前胡素对抗心肌梗死后心肌纤维化的作用研究

Effects of Decursin on Myocardial Fibrosis Following Myocardial Infarction

摘要

目的 探讨紫花前胡素（decursin，DEC）对小鼠心肌梗死后心肌纤维化的作用，及其对心肌成纤维细胞（cardiac fibroblasts，CFs）活化的调控机制。方法 选用C57BL/6J小鼠，行冠状动脉左前降支结扎术，构建小鼠心肌梗死模型，随机分组（每组 n = 5只），假手术组（Sham组）、心肌梗死模型组（MI组）、紫花前胡素治疗组（DEC组），通过灌胃给药的方式，给予DEC治疗，术后28 d，通过超声心动图评估心功能，免疫组织化学法检测心肌组织的病理形态变化，观察心肌纤维化程度；细胞划痕实验检测心肌成纤维细胞迁移能力；EdU技术检测心肌成纤维细胞增殖；Western blot检测心肌纤维化相关信号通路及关键蛋白表达水平。结果 超声心动图评估结果发现，DEC治疗组与MI组相比，左心室射血分数（LVEF，升高25%，P < 0.001）、左心室短轴缩短率（left ventricular ejection fraction，LVFS）升高16%，P < 0.001，二尖瓣E/A比值显著升高（MV E/A，P < 0.001）。免疫组化结果显示，MI组心肌间质胶原沉积增多、纤维化面积扩大（P < 0.001），而DEC干预可明显抑制纤维化进程（P < 0.01）。体外实验表明，DEC能显著抑制心肌成纤维细胞增殖（P < 0.01）与迁移（P < 0.05），使Fibronectin（P < 0.05）和α-SMA、Collagen I等蛋白表达下调（P < 0.01），并抑制心肌成纤维细胞的活化。结论 紫花前胡素具有改善心肌梗死后的心功能及抑制心肌纤维化的作用，其机制可能与PI3K/AKT信号通路相关。

Abstract

Objective To investigate the effects of Decursin (DEC) on myocardial fibrosis after myocardial infarction in mice and its regulatory mechanism underlying the activation of cardiac fibroblasts (CFs). Methods C57BL/6J mice were subjected to ligation of the left anterior descending coronary artery to establish a mouse myocardial infarction model. The mice were randomly divided into three groups (n = 5 per group): sham operation group (Sham), myocardial infarction model group (MI), and decursin treatment group (DEC). DEC was administered by intragastric gavage. For 28 days post-surgery, cardiac function was evaluated by echocardiography; histopathological changes and the degree of myocardial fibrosis were assessed by immunohistochemistry. The migratory capacity of cardiac fibroblasts was determined by wound-healing assay; the proliferation of cardiac fibroblasts was detected by EdU assay. The expression levels of key proteins related to myocardial fibrosis signaling pathways were measured by Western blot (WB). Results Echocardiographic assessment revealed that compared with the MI group, the DEC treatment group exhibited significantly increased left ventricular ejection fraction (LVEF, increased by 25%, P < 0.001), left ventricular fractional shortening (LVFS, increased by 16%, P < 0.001), and mitral valve E/A ratio (MV E/A, P < 0.001). Immunohistochemical results showed that the MI group presented increased myocardial interstitial collagen deposition and enlarged fibrotic area (P < 0.001), whereas DEC intervention markedly attenuated the fibrotic process (P < 0.01). In vitro experiments demonstrated that DEC significantly inhibited the proliferation (P < 0.01) and migration (P < 0.05) of cardiac fibroblasts, downregulated the expression of fibronectin (P < 0.05), α-SMA, collagen I and other proteins (P < 0.01), and suppressed the activation of cardiac fibroblasts. Conclusion Decursin can improve cardiac function and attenuate myocardial fibrosis after myocardial infarction, and its mechanism may be associated with the PI3K/AKT signaling pathway.

关键词

紫花前胡素 / 心肌梗死 / 心肌纤维化 / 成纤维细胞 / PI3K/AKT信号通路

Key words

Decursin / Myocardial infarction / Myocardial fibrosis / Fibroblasts / PI3K/AKT signaling pathway

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基金资助

国家自然科学基金(82070401)