The effect of miR-471-3p on macrophage polarization in the development of diabetic cardiomyopathy
The effect of miR-471-3p on macrophage polarization in the development of diabetic cardiomyopathy
复制标题
miR-471-3p对糖尿病心肌病发生过程中巨噬细胞极化的影响
DOI：
10.1016/j.lfs.2020.118989
复制
发表时间：
2021-01-18
期刊：
影响因子：
6.1
通讯作者：
Yuan, Qiong
中科院分区：
文献类型：
--
作者：
Liu, Guangqi;Yan, Dan;Yuan, Qiong
目的：M1/M2巨噬细胞比例失衡促进糖尿病心肌病（DCM）的发生，但其确切机制尚不完全清楚。本研究的目的是探讨miR-471-3p/沉默信息调节因子1（SIRT1）通路是否参与DCM发生过程中的巨噬细胞极化。方法：采用免疫组化染色检测心脏组织中M1和M2巨噬细胞的浸润情况。采用流式细胞术检测M1和M2巨噬细胞的比例。通过实时定量PCR对miR-471-3p的表达进行定量。将 miRNA 抑制剂转染至 RAW264.7 细胞中以研究其潜在机制。采用生物信息学方法和蛋白质印迹法探索miR-471-3p的靶基因，并通过双荧光素酶报告基因检测进一步证实。主要发现：我们观察到DCM组织心脏中M1型巨噬细胞浸润，而M2型巨噬细胞减少。 db/db 小鼠的骨髓源性巨噬细胞 (BMDM) 和经晚期糖基化终末产物 (AGE) 处理的 RAW264.7 细胞中，M1/M2 比率显着增加。同时，AGEs诱导的RAW264.7细胞中miR-471-3p显着上调，抑制miR-471-3p可以减少巨噬细胞的炎症极化。生物信息学分析确定 SIRT1 是 miR-471-3p 的靶标。双荧光素酶报告基因测定和蛋白质印迹均证实 miR-471-3p 负向调节 SIRT1 表达。 SIRT1激动剂白藜芦醇可以下调AGEs诱导的M1型巨噬细胞比例增加。结论：我们的研究结果表明，DCM的发生与AGEs诱导的巨噬细胞通过miR-471-3p/SIRT1通路极化为M1型相关。
Aims: The imbalance of M1/M2 macrophage ratio promotes the occurrence of diabetic cardiomyopathy (DCM), but the precise mechanisms are not fully understood. The aim of this study was to investigate whether miR-471-3p/silent information regulator 1 (SIRT1) pathway is involved in the macrophage polarization during the development of DCM.Methods: Immunohistochemical staining was used to detect M1 and M2 macrophages infiltration in the heart tissue. Flow cytometry was used to detect the proportion of M1 and M2 macrophages. Expression of miR-471-3p was quantified by real time quantitative-PCR. Transfection of miRNA inhibitor into RAW264.7 cells was performed to investigate the underlying mechanisms. Bioinformatics methods and western blotting were used to explore the target gene of miR-471-3p and further confirmed by dual luciferase reporter assay.Key findings: We observed that M1 macrophages infiltration in the heart of tissue in DCM while M2 type was decreased. M1/M2 ratio was increased significantly in bone marrow-derived macrophages (BMDMs) from db/db mice and in RAW264.7 cells treated with advanced glycation end products (AGEs). Meanwhile, miR-471-3p was significantly upregulated in RAW264.7 cells induced by AGEs and inhibition of miR-471-3p could reduce the inflammatory polarization of macrophages. Bioinformatics analysis identified SIRT1 as a target of miR-471-3p. Both dual luciferase reporter assay and western blotting verified that miR-471-3p negatively regulated SIRT1 expression. SIRT1 agonist resveratrol could downregulate the increased proportion of M1 macrophages induced by AGEs.Conclusion: Our results indicated that the development of DCM was related to AGEs-induced macrophage polarized to M1 type through a mechanism involving the miR-471-3p/SIRT1 pathway.