Sharma Dolly, Gupta Harshita, Gupta Avinash, Kumari Manisha, Varshney Rajeev, Meena Ramesh C
Department of Disruptive and Deterrence Technologies, Defence Institute of Physiology and Allied Sciences, Lucknow Road, Timarpur, Delhi, 110054 India.
3 Biotech. 2025 Apr;15(4):103. doi: 10.1007/s13205-025-04271-z. Epub 2025 Mar 31.
The present study was designed to investigate the effect of hypoxia (1% O) for 24 h in human AC16 cells by analyzing alterations in the expression of cardiac markers and signature pathways using immunocytochemistry and next-generation sequencing respectively. The Gene set enrichment analysis and Cytoscape software were used for data analysis and visualization respectively. Sequencing data validation and functional characterization were done using flow cytometry, qRT-PCR, an antibody array, and immunoblotting. The result revealed that the expression levels of troponins decreased; however, the expression levels of VEGF-A and HIF-alpha increased under hypoxia compared with unexposed control. A total of 2120 genes corresponding to 457 gene sets were significantly altered, 153 of which were significantly upregulated and 304 of which were downregulated in hypoxic cardiomyocytes. The significantly altered gene sets corresponded to key cellular and molecular pathways, such as cardiac hypertrophy, transcription factors, microRNAs, mitochondrial abnormalities, RNA processing, cell cycle, and biological oxidation pathways. Thus, this analysis revealed multiple pathways associated with hypoxia which provides valuable insights into the molecular mechanisms underlying human cardiomyocytes, identifying potential targets for addressing cardiac illnesses induced by hypoxia.
The online version contains supplementary material available at 10.1007/s13205-025-04271-z.
本研究旨在通过分别使用免疫细胞化学和下一代测序分析心脏标志物表达及特征性通路的变化，来研究缺氧（1%氧气）24小时对人AC16细胞的影响。基因集富集分析和Cytoscape软件分别用于数据分析和可视化。使用流式细胞术、qRT-PCR、抗体芯片和免疫印迹进行测序数据验证和功能表征。结果显示，肌钙蛋白的表达水平降低；然而，与未暴露的对照相比，缺氧条件下VEGF-A和HIF-α的表达水平升高。共有457个基因集对应的2120个基因发生了显著改变，其中153个在缺氧心肌细胞中显著上调，304个下调。显著改变的基因集对应于关键的细胞和分子通路，如心肌肥大、转录因子、微小RNA、线粒体异常、RNA加工、细胞周期和生物氧化途径。因此，该分析揭示了与缺氧相关的多种通路，为人类心肌细胞的分子机制提供了有价值的见解，确定了治疗缺氧诱导的心脏疾病的潜在靶点。
在线版本包含可在10.1007/s13205-025-04271-z获取的补充材料。
