Uric acid and sphingomyelin enhance autophagy in iPS cell-originated cardiomyocytes through lncRNA MEG3/miR-7-5p/EGFR axis
Uric acid and sphingomyelin enhance autophagy in iPS cell-originated cardiomyocytes through lncRNA MEG3/miR-7-5p/EGFR axis
复制标题
尿酸和鞘磷脂通过 lncRNA MEG3/miR-7-5p/EGFR 轴增强 iPS 细胞来源的心肌细胞的自噬
DOI：
10.1080/21691401.2019.1667817
复制
发表时间：
2019-12-04
影响因子：
5.8
通讯作者：
Huang, Guoying
中科院分区：
文献类型：
--
作者：
Cao, Yinyin;Wen, Junxiang;Huang, Guoying
本研究旨在确定与室间隔缺损(VSD)相关的代谢物及其潜在机制。采集VSD患者的血液和胸腺组织，分别进行基于LC-MS的代谢组学分析和iPS细胞来源的心肌细胞培养。体内实验采用VSD大鼠模型。采用RT-PCR、Western blotting、免疫组织化学、荧光素酶活性测定、GFP-LC3腺病毒和GFP及RFP tfLC3检测和透射电子显微镜等方法研究其作用机制。VSD患者血清中代谢产物尿酸(UA)和鞘磷脂(SM)升高，自噬增强。UA和SM联合应用可促进自噬，抑制EGFR和AKT3的表达。EGFR和AKT3的过表达分别抑制了UA和SM处理的心肌细胞的自噬。此外，通过靶向miR-7-5p，抑制和抑制UA和SM处理的心肌细胞的自噬。此外，miR-7-5p模拟物和抑制剂分别通过靶向EGFR促进和抑制UA和SM处理的心肌细胞的自噬。在VSD大鼠模型中，上调MEG3可以逆转高水平的自噬，降低血清UA和SM。综上所述，UA和SM是VSD相关代谢标志物，MEG3/miR-7-5p/EGFR轴在调节心肌细胞自噬中起关键作用。
This study aimed to determine the metabolites associated with ventricular septal defect (VSD) and the underlying mechanisms. Blood samples and thymus tissues were collected from VSD patients to perform LC-MS-based metabolomics assay and generate iPS cell-derived cardiomyocytes, respectively. VSD rat model was used in vivo study. RT-PCR, western blotting, immunohistochemistry, luciferase activity assay, GFP-LC3 adenovirus and GFP and RFP tfLC3 assay, and transmission electron microscopy were performed to investigate the underlying mechanisms. The metabolites uric acid (UA) and sphingomyelin (SM) increased in the serum of VSD patients, along with enhanced autophagy. The combination of UA and SM treatment could promote autophagy and inhibit EGFR and AKT3 expressions. Overexpression of EGFR and AKT3 suppressed autophagy in UA and SM-treated cardiomyocytes, respectively. Also, lncRNA MEG3 knockdown and overexpression could enhance and inhibit autophagy in UA and SM-treated cardiomyocytes, respectively, through targeting miR-7-5p. Moreover, miR-7-5p mimics and inhibitors promoted and inhibited autophagy in UA and SM-treated cardiomyocytes, respectively, via target EGFR. In VSD rat model, upregulation of MEG3 could reverse high level of autophagy and decrease serum UA and SM. In conclusion, UA and SM are essential VSD-associated metabolic biomarkers and MEG3/miR-7-5p/EGFR axis is critical to the regulation of autophagy in cardiomyocytes.