Faizo Nisreen, Narasimhulu Chandrakala Aluganti, Forsman Anna, Yooseph Shibu, Parthasarathy Sampath
Burnett School of Biomedical Sciences, Genomics and Bioinformatics Cluster, College of Medicine, University of Central Florida, Orlando, FL 32816, USA.
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL 32816, USA.
Foods. 2021 Feb 3;10(2):314. doi: 10.3390/foods10020314.
Lipid peroxides (LOOHs) abound in processed food and have been implicated in the pathology of diverse diseases including gut, cardiovascular, and cancer diseases. Recently, RNA Sequencing (RNA-seq) has been widely used to profile gene expression. To characterize gene expression and pathway dysregulation upon exposure to peroxidized linoleic acid, we incubated intestinal epithelial cells (Caco-2) with 100 µM of 13-hydroperoxyoctadecadienoic acid (13-HPODE) or linoleic acid (LA) for 24 h. Total RNA was extracted for library preparation and Illumina HiSeq sequencing. We identified 3094 differentially expressed genes (DEGs) in 13-HPODE-treated cells and 2862 DEGs in LA-treated cells relative to untreated cells. We show that 13-HPODE enhanced lipid metabolic pathways, including steroid hormone biosynthesis, PPAR signaling, and bile secretion, which alter lipid uptake and transport. 13-HPODE and LA treatments promoted detoxification mechanisms including cytochrome-P450. Conversely, both treatments suppressed oxidative phosphorylation. We also show that both treatments may promote absorptive cell differentiation and reduce proliferation by suppressing pathways involved in the cell cycle, DNA synthesis/repair and ribosomes, and enhancing focal adhesion. A qRT-PCR analysis of representative DEGs validated the RNA-seq analysis. This study provides insights into mechanisms by which 13-HPODE alters cellular processes and its possible involvement in mitochondrial dysfunction-related disorders and proposes potential therapeutic strategies to treat LOOH-related pathologies.
脂质过氧化物(LOOHs)在加工食品中大量存在,并与多种疾病的病理过程有关,包括肠道疾病、心血管疾病和癌症。最近,RNA测序(RNA-seq)已被广泛用于分析基因表达。为了表征暴露于过氧化亚油酸后基因表达和通路失调情况,我们将肠上皮细胞(Caco-2)与100µM的13-氢过氧化十八碳二烯酸(13-HPODE)或亚油酸(LA)孵育24小时。提取总RNA用于文库制备和Illumina HiSeq测序。相对于未处理的细胞,我们在13-HPODE处理的细胞中鉴定出3094个差异表达基因(DEGs),在LA处理的细胞中鉴定出2862个DEGs。我们发现13-HPODE增强了脂质代谢途径,包括类固醇激素生物合成、PPAR信号传导和胆汁分泌,这些途径会改变脂质摄取和运输。13-HPODE和LA处理促进了包括细胞色素P45在内的解毒机制。相反,两种处理均抑制了氧化磷酸化。我们还表明,两种处理可能通过抑制参与细胞周期、DNA合成/修复和核糖体的途径,并增强粘着斑,从而促进吸收性细胞分化并减少增殖。对代表性DEGs的qRT-PCR分析验证了RNA-seq分析。本研究深入探讨了13-HPODE改变细胞过程的机制及其可能参与线粒体功能障碍相关疾病的情况,并提出了治疗与LOOH相关病理的潜在治疗策略。