Growth differentiation factor 11 promotes macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway
Growth differentiation factor 11 promotes macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway
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DOI：
10.2478/fzm-2023-0008
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发表时间：
2023-01
期刊：
影响因子：
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通讯作者：
Manyu Gong;Xuewen Yang;Yaqi Wang;Yanying Wang;Dongping Liu;H. Li;Yu-Chun Qu;Xiyang Zhang
中科院分区：
文献类型：
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作者：
Manyu Gong;Xuewen Yang;Yaqi Wang;Yanying Wang;Dongping Liu;H. Li;Yu-Chun Qu;Xiyang Zhang
心肌梗死（MI）是一种严重威胁人类健康的疾病，尤其是在寒冷环境中。巨噬细胞向不同功能表型（M1巨噬细胞和M2巨噬细胞）的极化与MI修复密切相关。生长分化因子11（GDF 11）在炎症相关疾病中起重要作用。在这项研究中，我们研究了GDF 11在巨噬细胞极化中的调节作用，并阐明了MI的潜在机制。方法采用永久性结扎冠状动脉左前降支（LAD）建立小鼠心肌梗死模型，将小鼠随机分为假手术组、心肌梗死组和心肌梗死+GDF 11组。通过超声心动图、氯化三苯基四氮唑染色和心脏组织免疫荧光染色证实GDF 11对心肌梗死的保护作用及其对巨噬细胞极化的影响。在体外，基于RAW264.7细胞系，通过qRT-PCR、蛋白质印迹和流式细胞术验证了GDF 11通过抑制Notch 1信号通路促进巨噬细胞向M2型极化的作用。结果GDF 11在MI小鼠心肌组织中表达明显下调。补充GDF 11可改善心功能。GDF 11可降低MI小鼠心肌组织中M1巨噬细胞的比例，增加M2巨噬细胞的聚集。巨噬细胞清除后，GDF 11对MI小鼠的心脏保护作用减弱。在细胞水平，应用GDF 11可以以剂量依赖性方式抑制M1巨噬细胞（经典活化的巨噬细胞）标志物iNOS、白细胞介素（IL）-1β和IL-6的表达。相反，GDF 11显著增加M2巨噬细胞标志物的水平，包括IL-10、CD 206、精氨酸酶1（Arg 1）和血管内皮生长因子（VEGF）。有趣的是，GDF 11可以促进M1巨噬细胞极化为M2巨噬细胞。在分子水平上，GDF 11显著下调Notch 1信号通路，其激活已被证明促进巨噬细胞中的M1极化。结论GDF 11通过抑制Notch 1信号通路促进巨噬细胞向M2极化，减轻心肌梗死。
Abstract Background Myocardial infarctions (MI) is a major threat to human health especially in people exposed to cold environment. The polarization of macrophages towards different functional phenotypes (M1 macrophages and M2 macrophages) is closely related to MI repairment. The growth differentiation factor 11 (GDF11) has been reported to play a momentous role in inflammatory associated diseases. In this study, we examined the regulatory role of GDF11 in macrophage polarization and elucidated the underlying mechanisms in MI. Methods In vivo, the mice model of MI was induced by permanent ligation of the left anterior descending coronary artery (LAD), and mice were randomly divided into the sham group, MI group, and MI+GDF11 group. The protective effect of GDF11 on myocardial infarction and its effect on macrophage polarization were verified by echocardiography, triphenyl tetrazolium chloride staining and immunofluorescence staining of heart tissue. In vitro, based on the RAW264.7 cell line, the effect of GDF11 in promoting macrophage polarization toward the M2 type by inhibiting the Notch1 Signaling pathway was validated by qRT-PCR, Western blot, and flow cytometry. Results We found that GDF11 was significantly downregulated in the cardiac tissue of MI mice. And GDF11 supplementation can improve the cardiac function. Moreover, GDF11 could reduce the proportion of M1 macrophages and increase the accumulation of M2 macrophages in the heart tissue of MI mice. Furthermore, the cardioprotective effect of GDF11 on MI mice was weakened after macrophage clearance. At the cellular level, application of GDF11 could inhibit the expression of M1 macrophage (classically activated macrophage) markers iNOS, interleukin (IL)-1β, and IL-6 in a dose-dependent manner. In contrast, GDF11 significantly increased the level of M2 macrophage markers including IL-10, CD206, arginase 1 (Arg1), and vascular endothelial growth factor (VEGF). Interestingly, GDF11 could promote M1 macrophages polarizing to M2 macrophages. At the molecular level, GDF11 significantly down-regulated the Notch1 signaling pathway, the activation of which has been demonstrated to promote M1 polarization in macrophages. Conclusions GDF11 promoted macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway.