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Long non-coding RNA and mRNA analysis of Ang II-induced neuronal dysfunction

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Abstract

The sustained activation of Angiotensin II (Ang II) induces the remodelling of neurovascular units, inflammation and oxidative stress reactions in the brain. Long non-coding RNAs (lncRNAs) play a crucial regulatory role in the pathogenesis of hypertensive neuronal damage. The present study aimed to substantially extend the list of potential candidate genes involved in Ang II-related neuronal damage. This study assessed apoptosis and energy metabolism with Annexin V/PI staining and a Seahorse assay after Ang II exposure in SH-SY5Y cells. The expression of mRNA and lncRNA was investigated by transcriptome sequencing. The integrated analysis of mRNA and lncRNAs and the molecular mechanism of Ang II on neuronal injury was analysed by bioinformatics. Ang II increased the apoptosis rate and reduced the energy metabolism of SH-SY5Y cells. The data showed that 702 mRNAs and 821 lncRNAs were differentially expressed in response to Ang II exposure (244 mRNAs and 432 lncRNAs were upregulated, 458 mRNAs and 389 lncRNAs were downregulated) (fold change ≥ 1.5, P < 0.05). GO and KEGG analyses showed that both DE mRNA and DE lncRNA were enriched in the metabolism, differentiation, apoptosis and repair of nerve cells. This is the first report of the lncRNA–mRNA integrated profile of SH-SY5Y cells induced by Ang II. The novel targets revealed that the metabolism of the vitamin B group, the synthesis of unsaturated fatty acids and glycosphingolipids are involved in the Ang II-related cognitive impairment. Sphingolipid metabolism, the Hedgehog signalling pathway and vasopressin-regulated water reabsorption play important roles in nerve damage.

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Abbreviations

Ang II:

Angiotensin II

lncRNA:

Long non-coding RNA

SH-SY5Y:

Human nerve cell

DMEM/F12:

Dulbecco’s modified eagle medium: nutrient mixture F-12

FBS:

Fetal bovine serum

OCR:

Oxygen consumption rate

DE:

Differential expression

FC:

Fold change

GO:

GeNE ontology

BP:

Biological process

CC:

Cellular components

MF:

Molecular function

ACE2:

Angiotensin converting enzyme 2

LRP2:

Low-density lipoprotein receptor-related protein 2

ECEL1:

Endothelin-converting enzyme-like 1

VIP:

Vasoactive intestinal peptide

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Acknowledgements

This study was supported by National Natural Science Foundation of China No. 81573916 and Shandong Province ‘Taishan Scholar’ Construction Project Funds No. 2018-35. We thank Yixin Yin of Shanghai Biotechnology Corporation for technical assistance and constructive suggestions.

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Authors and Affiliations

  1. First Clinical Medical College, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China

    Lin-Lin Shao & Ying-Zi Qi

  2. Department of Cardiovascular, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, No. 16369, Jingshi Road, Jinan, 250011, China

    Yue-Hua Jiang, Ling-Yu Jiang & Chuan-Hua Yang

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Contributions

LLS, performed the experiments and drafted the manuscript. YHJ designed the study and performed the experiments. LYJ carried out methodology and data analysis. CHY designed the study, contributed to discussion and carried out important revisions of the article. YZQ contributed to data curation and software applications. All authors read and approved the final manuscript.

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Correspondence to Chuan-Hua Yang.

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Shao, LL., Jiang, YH., Jiang, LY. et al. Long non-coding RNA and mRNA analysis of Ang II-induced neuronal dysfunction. Mol Biol Rep 46, 3233–3246 (2019). https://doi.org/10.1007/s11033-019-04783-x

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