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牦牛 PASMCs 在低氧和常氧条件下的联合转录组和蛋白质组分析。

Combined transcriptome and proteome analysis of yak PASMCs under hypoxic and normoxic conditions.

机构信息

Life Science and Engineering College, Northwest Minzu University, Lan, China.

Biomedical Research Center, Northwest Minzu University, Lan Zhou, China.

出版信息

PeerJ. 2022 Nov 25;10:e14369. doi: 10.7717/peerj.14369. eCollection 2022.

Abstract

BACKGROUND

Yaks are animals that have lived in plateau environments for generations. Yaks can adapt to the hypoxic plateau environment and also pass this adaptability on to the next generation. The lungs are the most important respiratory organs for mammals to adapt to their environment. Pulmonary artery smooth muscle cells play an important role in vascular remodeling under hypoxia, but the genetic mechanism underpinning the yak's ability to adapt to challenging plateau conditions is still unknown.

METHODS

A tandem mass tag (TMT) proteomics study together with an RNA-seq transcriptome analysis were carried out on pulmonary artery smooth muscle cells (PASMCs) that had been grown for 72 hours in both normoxic (20% O2) and hypoxic (1% O2) environments. RNA and TP (total protein) were collected from the hypoxic and normoxic groups for RNA-seq transcriptome sequencing and TMT marker protein quantification, and RT-qPCR validation was performed.

RESULTS

A total of 17,711 genes and 6,859 proteins were identified. Further, 5,969 differentially expressed genes (DEGs) and 531 differentially expressed proteins (DEPs) were identified in the comparison group, including 2,924 and 186 upregulated genes and proteins and 3,045 and 345 down-regulated genes and proteins, respectively. The transcriptomic and proteomic analyses revealed that 109 DEGs and DEPs were highly positively correlated, with 77 genes showing the same expression trend. Nine overlapping genes were identified in the HIF-1 signaling pathway, glycolysis / gluconeogenesis, central carbon metabolism in cancer, PPAR signaling pathway, AMPK signaling pathway, and cholesterol metabolism (PGAM1, PGK1, TPI1, HMOX1, IGF1R, OLR1, SCD, FABP4 and LDLR), suggesting that these differentially expressed genes and protein functional classifications are related to the hypoxia-adaptive pathways. Overall, our study offers abundant data for further analysis of the molecular mechanisms in yak PASMCs and their adaptability to different oxygen concentrations.

摘要

背景

牦牛是世代生活在高原环境中的动物。它们能够适应低氧的高原环境,并将这种适应性遗传给下一代。肺是哺乳动物适应环境最重要的呼吸器官。肺动脉平滑肌细胞在低氧环境下的血管重构中起着重要作用,但牦牛适应挑战性高原环境的遗传机制尚不清楚。

方法

对在常氧(20% O2)和低氧(1% O2)环境中培养 72 小时的肺动脉平滑肌细胞(PASMCs)进行串联质量标签(TMT)蛋白质组学研究和 RNA-seq 转录组分析。从低氧和常氧组中收集 RNA 和 TP(总蛋白)进行 RNA-seq 转录组测序和 TMT 标记蛋白定量,并进行 RT-qPCR 验证。

结果

共鉴定出 17711 个基因和 6859 个蛋白质。进一步在比较组中鉴定出 5969 个差异表达基因(DEGs)和 531 个差异表达蛋白(DEPs),分别包括 2924 个和 186 个上调基因和蛋白,以及 3045 个和 345 个下调基因和蛋白。转录组和蛋白质组分析表明,109 个 DEGs 和 DEPs 高度正相关,其中 77 个基因具有相同的表达趋势。在 HIF-1 信号通路、糖酵解/糖异生、癌症中心碳代谢、PPAR 信号通路、AMPK 信号通路和胆固醇代谢(PGAM1、PGK1、TPI1、HMOX1、IGF1R、OLR1、SCD、FABP4 和 LDLR)中发现了 9 个重叠基因,表明这些差异表达基因和蛋白的功能分类与低氧适应途径有关。总的来说,我们的研究为进一步分析牦牛 PASMCs 的分子机制及其对不同氧浓度的适应性提供了丰富的数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d55c/9703989/c49392ba0bf0/peerj-10-14369-g001.jpg

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