Guo Yingying, Cen Xian-Feng, Li Dan, Qiu Hong-Liang, Chen Ya-Jie, Zhang Meng, Huang Si-Hui, Xia Hao, Xu Man
Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, China.
Hubei Key Laboratory of Metabolic and Chronic Diseases, Renmin Hospital of Wuhan University, Wuhan, China.
Front Cardiovasc Med. 2023 Jun 19;10:1137429. doi: 10.3389/fcvm.2023.1137429. eCollection 2023.
Chronic pressure overload triggers pathological cardiac hypertrophy that eventually leads to heart failure. Effective biomarkers and therapeutic targets for heart failure remain to be defined. The aim of this study is to identify key genes associated with pathological cardiac hypertrophy by combining bioinformatics analyses with molecular biology experiments.
Comprehensive bioinformatics tools were used to screen genes related to pressure overload-induced cardiac hypertrophy. We identified differentially expressed genes (DEGs) by overlapping three Gene Expression Omnibus (GEO) datasets (GSE5500, GSE1621, and GSE36074). Correlation analysis and BioGPS online tool were used to detect the genes of interest. A mouse model of cardiac remodeling induced by transverse aortic constriction (TAC) was established to verify the expression of the interest gene during cardiac remodeling by RT-PCR and western blot. By using RNA interference technology, the effect of transcription elongation factor A3 (Tcea3) silencing on PE-induced hypertrophy of neonatal rat ventricular myocytes (NRVMs) was detected. Next, gene set enrichment analysis (GSEA) and the online tool ARCHS4 were used to predict the possible signaling pathways, and the fatty acid oxidation relevant pathways were enriched and then verified in NRVMs. Furthermore, the changes of long-chain fatty acid respiration in NRVMs were detected using the Seahorse XFe24 Analyzer. Finally, MitoSOX staining was used to detect the effect of Tcea3 on mitochondrial oxidative stress, and the contents of NADP(H) and GSH/GSSG were detected by relevant kits.
A total of 95 DEGs were identified and Tcea3 was negatively correlated with Nppa, Nppb and Myh7. The expression level of Tcea3 was downregulated during cardiac remodeling both and . Knockdown of Tcea3 aggravated cardiomyocyte hypertrophy induced by PE in NRVMs. GSEA and online tool ARCHS4 predict Tcea3 involved in fatty acid oxidation (FAO). Subsequently, RT-PCR results showed that knockdown of Tcea3 up-regulated Ces1d and Pla2g5 mRNA expression levels. In PE induced cardiomyocyte hypertrophy, Tcea3 silencing results in decreased fatty acid utilization, decreased ATP synthesis and increased mitochondrial oxidative stress.
Our study identifies Tcea3 as a novel anti-cardiac remodeling target by regulating FAO and governing mitochondrial oxidative stress.
慢性压力超负荷引发病理性心脏肥大，最终导致心力衰竭。心力衰竭有效的生物标志物和治疗靶点仍有待确定。本研究旨在通过生物信息学分析与分子生物学实验相结合，鉴定与病理性心脏肥大相关的关键基因。
使用综合生物信息学工具筛选与压力超负荷诱导的心脏肥大相关的基因。通过重叠三个基因表达综合数据库（GEO）数据集（GSE5500、GSE1621和GSE36074）鉴定差异表达基因（DEG）。使用相关性分析和BioGPS在线工具检测感兴趣的基因。建立经主动脉缩窄（TAC）诱导的心脏重塑小鼠模型，通过RT-PCR和蛋白质印迹法验证心脏重塑过程中感兴趣基因的表达。利用RNA干扰技术，检测转录延伸因子A3（Tcea3）沉默对苯肾上腺素（PE）诱导的新生大鼠心室肌细胞（NRVM）肥大的影响。接下来，使用基因集富集分析（GSEA）和在线工具ARCHS4预测可能的信号通路，富集脂肪酸氧化相关通路并在NRVM中进行验证。此外，使用海马XFe24分析仪检测NRVM中长链脂肪酸呼吸的变化。最后，使用MitoSOX染色检测Tcea3对线粒体氧化应激的影响，并通过相关试剂盒检测烟酰胺腺嘌呤二核苷酸磷酸（NADP(H)）和还原型谷胱甘肽/氧化型谷胱甘肽（GSH/GSSG）的含量。
共鉴定出95个DEG，Tcea3与利钠肽A（Nppa）、利钠肽B（Nppb）和肌球蛋白重链7（Myh7）呈负相关。在心脏重塑过程中，Tcea3的表达水平在[此处原文缺失具体阶段信息]均下调。Tcea3的敲低加重了PE诱导的NRVM肥大。GSEA和在线工具ARCHS4预测Tcea3参与脂肪酸氧化（FAO）。随后，RT-PCR结果显示，Tcea3的敲低下调了羧酸酯酶1D（Ces1d）和磷脂酶A2，Group V（Pla2g5）mRNA表达水平。在PE诱导的心肌细胞肥大中，Tcea3沉默导致脂肪酸利用减少、ATP合成减少和线粒体氧化应激增加。
我们的研究通过调节FAO和控制线粒体氧化应激，将Tcea3鉴定为一个新的抗心脏重塑靶点。
