RNA 测序揭示了与葛根素介导的小鼠心肌肥厚抑制相关的新型 LncRNA/mRNAs 共表达网络。
RNA sequencing reveals novel LncRNA/mRNAs co-expression network associated with puerarin-mediated inhibition of cardiac hypertrophy in mice.
机构信息
Department of Cardiology, Laboratory of Heart Center, Guangdong Provincial Biomedical Engineering Technology Research Center for Cardiovascular Disease, Sino-Japanese Cooperation Platform for Translational Research in Heart Failure, Zhujiang Hospital, Southern Medical University, Guangzhou, PR China.
Department of Geriatrics, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
出版信息
PeerJ. 2022 Apr 5;10:e13144. doi: 10.7717/peerj.13144. eCollection 2022.
BACKGROUND
Evidence has demonstrated that puerarin is a potential medicine for the treatment of cardiac hypertrophy. However, the precise underlying molecular mechanisms of the protective effect of puerarin are still unclear. Here, we aimed to explore the regulatory mechanisms of lncRNAs/mRNAs co-expression network in a cardiac hypertrophy mouse model after puerarin treatment.
METHODS
A mouse model of cardiac hypertrophy was established by transverse aortic constriction (TAC). The echocardiography, tissue staining and western blot were used to examine the protective effect of puerarin. Then RNA sequencing (RNA-seq) was carried out to analyze systematically mRNAs and lncRNAs expression. The target lncRNA were confirmed using qRT-PCR. Moreover, a coding/non-coding gene co-expression network were established to find the interaction of lncRNA and mRNAs. The biological process, cellular component, molecular function and pathways of different expression mRNAs targeted by lncRNA were explored using Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis.
RESULTS
Puerarin exhibited an obvious inhibitory effect in cardiac hypertrophy in TAC model. RNA-seq analysis was performed to investigate the lncRNAs and mRNAs expression patterns of cardiomyocytes in sham and TAC groups treated with or without puerarin. RNA-seq identified that TAC downregulated four lncRNAs, which could be revised by puerarin treatment (|log2 Fold change| > 2 and FDR < 0.05). Among them, expression alterations of lncRNA Airn (antisense of Igf2r non-protein coding RNA) was confirmed by qRT-PCR. Pearson's correlation coefficients of co-expression levels suggested that there was an interactive relationship between Airn and 2,387 mRNAs ( > 0.95 or < -0.95). Those co-expressed mRNAs were enriched in some important biological processes such as translational initiation, cell proliferation, insulin-like growth factor binding and poly(A) RNA binding. KEGG analyses suggested that those Airn-interacted mRNAs were enriched in endocytosis, signaling pathways regulating pluripotency of stem cells and the Jak-STAT pathway.
CONCLUSION
Puerarin may exert beneficial effects on cardiac hypertrophy through regulating the lncRNAs/mRNAs co-expression network.
背景
有证据表明，葛根素是治疗心肌肥厚的潜在药物。然而，葛根素保护作用的确切潜在分子机制尚不清楚。在这里，我们旨在探讨葛根素处理后心肌肥厚小鼠模型中长链非编码 RNA（lncRNA）/mRNA 共表达网络的调控机制。
方法
通过横主动脉缩窄（TAC）建立心肌肥厚小鼠模型。采用超声心动图、组织染色和 Western blot 检测葛根素的保护作用。然后进行 RNA 测序（RNA-seq）以系统分析 mRNAs 和 lncRNAs 的表达。采用 qRT-PCR 验证靶 lncRNA。此外，建立编码/非编码基因共表达网络，以发现 lncRNA 与 mRNAs 的相互作用。通过基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）途径分析，探讨不同表达 mRNAs 靶向的 lncRNA 的生物学过程、细胞成分、分子功能和途径。
结果
葛根素对 TAC 模型中的心肌肥厚有明显的抑制作用。进行 RNA-seq 分析，以研究心肌细胞在 sham 和 TAC 组中 lncRNAs 和 mRNAs 的表达模式，以及有无葛根素处理的情况。RNA-seq 鉴定出 TAC 下调了 4 个 lncRNA，这些 lncRNA可以被葛根素治疗所修正（|log2 Fold change| > 2，FDR < 0.05）。其中，lncRNA Airn（Igf2r 非蛋白编码 RNA 的反义）的表达变化通过 qRT-PCR 得到证实。共表达水平的 Pearson 相关系数表明，Airn 与 2387 个 mRNAs 之间存在相互作用关系（>0.95 或 < -0.95）。这些共表达的 mRNAs富集在一些重要的生物学过程中，如翻译起始、细胞增殖、胰岛素样生长因子结合和 poly（A）RNA 结合。KEGG 分析表明，Airn 相互作用的 mRNAs 富集在内吞作用、调节干细胞多能性的信号通路和 Jak-STAT 通路。
结论
葛根素可能通过调节 lncRNAs/mRNAs 共表达网络对心肌肥厚发挥有益作用。
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